Microbial Biochemistry

CA Selects: Fermentation Chemicals
Issue 3, 2000
Page 1
MICROBIAL BIOCHEMISTRY
132: 20857g Methylsulfomycin I, a new cyclic peptide antibiotic from Streptomyces sp. HIL Y-9420704. Kumar, E. K. S.
Vijaya; Kenia, J.; Mukhopadhyay, Triptikumar; Nadkarni, S. R.
(Research Centre, Hoechst Marion Roussel Limited, Mumbai, 400
080 India). J. Nat. Prod. 1999, 62(11), 1562-1564 (Eng), American
Chemical Society. Methylsulfomycin I (I) is a new cyclic peptide
CH2
O
N
H
NH2
N
H
CH2
CH2
O
H
N
Me
N
O
HO
Me
NH
CH2
HN
N
H
OH
Me
H2 C
NH
N
Me
O
NH
O
MeO
H
N
N
S
Me
Me
antibiotic isolated from the fermn. broth of a Streptomyces sp. HIL

Y-9420704. Its structure was elucidated by NMR and GC-MS. The
in vitro activity (MIC) against a wide range of Gram-pos. bacteria,
including methicillin-resistant Staphylococcus aureus and vancomycin-, and teicoplanin-resistant strains, is described.
132: 20858h Isolation and characterization of a novel antifungal peptide from Aspergillus niger. Gun Lee, Dong; Yub Shin,
Song; Maeng, Cheol-Young; Zhu Jin, Zhe; Lyong Kim, Kil; Hahm,
Kyung-Soo (Peptide Engineering Research Unit, Korea Research
Institute of Bioscience and Biotechnology, Taejon, S. Korea). Biochem. Biophys. Res. Commun. 1999, 263(3), 646-651 (Eng), Academic
Press. A novel antifungal peptide (termed as Anafp) was isolated
from the culture supernatant of the filamentous fungi, Aspergillus
niger. The whole amino acid sequence of Anafp was detd. and the
peptide was found to be composed of a single polypeptide chain with
58 amino acids including six cysteine residues. The peptide shows
some degree of sequence homol. to a cysteine-rich antifungal peptides reported from the seeds of Sinapis alba and Arabidopsis thaliana
or the extracellular media of Aspergillus giganteus and Penicillium
chrysogenumsome. Cysteine-spacing pattern of Anafp was similar
to that of the antifungal peptide from Penicillium chrysogenum. The
Anafp exhibited potent growth inhibitory activities against yeast
strains as well as filamentous fungi at a range from 4 to 15 M. In
contrast, Anafp did not show antibacterial activity against Escherichia coli and Bacillus subtilis even at 50 M. (c) 1999 Academic
Press.
132: 20860c Novel radicinol derivatives from long-term cultures of Alternaria chrysanthemi. Sheridan, Helen; Canning,
Ann-Marie (Department of Pharmacognosy School of Pharmacy,
Trinity College, Dublin, Ire.). J. Nat. Prod. 1999, 62(11), 1568-1569
(Eng), American Chemical Society. Cultures of Alternaria chrysanO
R1
against oral pathogens. Groenink, J.; Walgreen-Weterings, E.;

van 't Hof, W.; Veerman, E. C. I.; Nieuw Amerongen, A. V. (Section
Oral Biochemistry, Department of Oral Biology, Academic Centre for
Dentistry Amsterdam (ACTA), 1081 BT Amsterdam, Neth.). FEMS
Microbiol. Lett. 1999, 179(2), 217-222 (Eng), Elsevier Science B.V..
Peptides derived from the N-terminal domain that comprises an amphipathic R-helix in human lactoferrin (LFh 18-31 and LFh 20-38)
and bovine lactoferrin (LFb 17-30 and LFb 19-37) were chem.
synthesized. Since many pos. charged amphipathic R-helixes contain
antimicrobial activity, the peptides were tested for their antimicrobial
activity against various oral pathogens. Both peptides from bovine
lactoferrin had more potent antimicrobial activities than the human
equiv. Peptide LFb 17-30, contg. the largest no. of pos. charged
amino acids, showed the highest antimicrobial activity to both Grampos. and Gram-neg. bacteria. Since native lactoferrin mols. had no
killing activity, release of these peptides from the native protein should
be investigated to explore the use in oral care products.
132: 20867k UV-guided isolation of verrucines A and B, novel
quinazolines from Penicillium verrucosum structurally related
to anacine from Penicillium aurantiogriseum. Larsen, Thomas
Ostenfeld; Franzyk, Henrik; Jensen, Soren Rosendal (Mycology
Group Department of Biotechnology and Department of Organic
Chemistry, Technical University of Denmark, Lyngby, 2800 Den.). J.
Nat. Prod. 1999, 62(11), 1578-1580 (Eng), American Chemical
Society. Two novel quinazolines, verrucines A (I) and B (II), have
H
N
NH
N
O
H
O
O
I R,R1 =b-H,b-H
II R,R1 =a-H,b-H or b-H,a-H
NH2
Me
Me
H
N
NH
N
O
H
O
O
NH2
III
been isolated as a major and a minor metabolite, resp., of Penicillium

verrucosum. Both are condensates of one mole each of anthranilic
acid, phenylalanine, and glutamine. The structures were elucidated
by spectroscopic methods, and the two compds. were found to be
epimers. The spectroscopic data for the ostensible benzodiazepine
anacine reported from Penicillium aurantiogriseum, composed of one
mole each of anthranilic acid, leucine, and glutamine, appeared very
similar to those of I. Therefore, a revised quinazoline structure for
anacine (III) is proposed.
OH
O
O
Me
Me
O
I R=H, R1 =OH
II R=OH, R1 =H
III R=OMe, R1 =H
OMe
OH
O
O
Me
Me
IV
themi normally produce radicinin and radicinol (I) when cultured on

Czapek-Dox medium or on potato dextrose broth. We have obsd.
that long-term cultures of A. chrysanthemi grown on malt-ext. broth
produce 3-epiradicinol (II), the novel metabolites 3-methoxy-3epiradicinol (III) and 9,10-epoxy-3-methoxy-3-epiradicinol (IV),
and (I).
132: 20863f Cationic amphipathic peptides, derived from
bovine and human lactoferrins, with antimicrobial activity
132: 20869n Characterization of mutations in the rpoB gene

in naturally rifampin-resistant Rickettsia species. Drancourt,
Michel; Raoult, Didier (Unite des Rickettsies CNRS UPRES-A 6020,
Faculte de Medecine, Universite de la Mediterranee, 13385 Marseille,
Fr.). Antimicrob. Agents Chemother. 1999, 43(10), 2400-2403 (Eng),
American Society for Microbiology. Rickettsiae are gram-neg., obligately intracellular bacteria responsible for arthropod-borne spotted
fevers and typhus. Exptl. studies have delineated a cluster of naturally rifampin-resistant spotted fever group species. We sequenced
the 4122-4125-bp RNA polymerase -subunit-encoding gene (rpoB)
from typhus and spotted fever group representatives and obtained
partial sequences for all naturally rifampin-resistant species. A
single point mutation resulting in a phenylalanine-to-leucine change
at position 973 of the Rickettsia conorii rpoB sequence and present
in all the rifampin-resistant species was absent in all the rifampinsusceptible species. RpoB-based phylogenetic relationships among
Page 2
these rickettsial species yielded topologies which were in accordance

with previously published phylogenies.
132: 20871g Lyngbyastatin 2 and norlyngbyastatin 2, analogues of dolastatin G and nordolastatin G from the marine
cyanobacterium Lyngbya majuscula. Luesch, Hendrik; Yoshida,
Wesley Y.; Moore, Richard E.; Paul, Valerie J. (Department of
Chemistry, University of Hawaii at Manoa, Honolulu, HI 96822 USA).
J. Nat. Prod. 1999, 62(12), 1702-1706 (Eng), American Chemical
Society. Lyngbyastatin 2 (I) and norlyngbyastatin 2 (II), new cyto-
Me
OMe
Me
Me
Me
Me
Me
N
H
O
Me
Me
Me
N
O
O
OH
Me
Me
Me
Me
O
Me
OMe
Me
Me
Me
Me
OMe
Me
Me
Me
N
H
O
Me
Me
Me
N
O
O
O
OH
Me
Me
Me
O
Me
O
O
Me
Me
II
toxic analogs of dolastatin G and nordolastatin G, resp., have been

isolated as constituents from a Guamanian variety of the marine
cyanobacterium Lyngbya majuscula. Structure elucidation of these
cyclic depsipeptides relied on extensive application of 2D NMR
techniques. The finding of these new metabolites further supports
the proposal that many compds. originally isolated from the sea hare
Dolabella auricularia are most likely of cyanobacterial origin.
132: 20876n Immunomodulatory constituents from an ascomycete, Microascus tardifaciens. Fujimoto, Haruhiro; Fujimaki,
Toshiyuki; Okuyama, Emi; Yamazaki, Mikio (Faculty of Pharmaceutical Sciences, Chiba University, Chiba, Japan 263-8522). Chem.
Pharm. Bull. 1999, 47(10), 1426-1432 (Eng), Pharmaceutical Society
of Japan. Fractionation guided by the immunosuppressive activity of
the defatted AcOEt ext. of an Ascomycete, Microascus tardifaciens,
afforded eight constituents, questin (emodin 8-O-methylether) (I),
rubrocristin (II), 5,7-dihydroxy-4-methylphthalide (III), cladosporin
(asperentin) (IV), cladosporin 8-O-methylether (V), tardioxopiperazine A [cyclo-L-alanyl-5-isopentenyl-2-(1',1'-dimethylallyl)-Ltryptophan] (VI), tardioxopiperazine B [cyclo-L-alanyl-7-isopentenyl-2-(1',1'-dimethylallyl)-L-tryptophan] (VII), and asperflavin
(VIII), among which VI and VII were new compds. Compds. I and
II showed considerably high immunosuppressive activity, VI was
moderate and, III, IV, V, VII, and VIII showed low activity.
132: 20877p Cytokinin production by Paenibacillus polymyxa. Timmusk, S.; Nicander, B.; Granhall, U.; Tillberg, E.
(Department of Plant Biology, Swedish University of Agricultural Sciences, S-750 07 Uppsala, Swed.). Soil Biol. Biochem. 1999, 31(13),
1847-1852 (Eng), Elsevier Science Ltd.. The prodn. of hormones
has been suggested to be one of the mechanisms by which plant
growth-promoting rhizobacteria (PGPR) stimulate plant growth. To
evaluate whether the free-living soil bacterium, Paenibacillus polymyxa, releases the hormone group cytokinins and, if so, their identity,
the content of cytokinins in the growth media, before and after cultivation of this bacterium, was detd. by immunoaffinity chromatog. (IAC).
This method allows the isolation of almost all known cytokinins and
their metabolites. Sepn. and characterization were done by high
performance liq. chromatog. (HPLC) with online UV detection, and
final identification was by gas chromatog.-mass spectrometry. Isopentenyladenine (iP) was identified in the two defined media used for
the cultivation of P. polymyxa, but not earlier than at its late stationary growth. A third medium, supplemented with yeast ext., contained
iso-pentenyladenine riboside (iPR) and some addnl. cytokinin-like
substances before inoculation. When the same medium was sampled
Issue 3, 2000
after the cultivation of P. polymyxa up to its logarithmic growth phase,

the cytokinin concn. had decreased. After prolonged cultivation of P.
polymyxa, small amts. of iP appeared in all three media, and iPR had
disappeared from the yeast-contg. medium, which indicates that the
bacterium can metabolize cytokinins.
132: 20878q Palmitoylation of the intracytoplasmic R peptide of the transmembrane envelope protein in Moloney murine leukemia virus. Olsen, Katharina E. P.; Andersen, Klaus B.
(Department of Pharmacology, The Royal Danish School of Pharmacy,
Copenhagen, DK-2100 Den.). J. Virol. 1999, 73(11), 8975-8981
(Eng), American Society for Microbiology. Previously it was reported
that the 16-amino-acid (aa) C-terminal cytoplasmic tail of Moloney
murine leukemia virus (MoMLV) transmembrane protein Pr15E is
cleaved off during virus synthesis, yielding the mature, fusion active
transmembrane protein p15E and the 16-aa peptide (R peptide or
p2E). It remains to be elucidated how the R peptide impairs fusion
activity of the uncleaved Pr15E. The R peptide from MoMLV was
analyzed by Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunostained with antiserum against the synthetic
16-aa R peptide. The R peptide resolved with an apparent mol.
mass of 7 kDa and not the 4 kDa seen with the corresponding synthetic
peptide. The 7-kDa R peptide was found to be membrane bound in
MoMLV-infected NIH 3T3 cells, showing that cleavage of the 7-kDa
R-peptide tail must occur before or during budding of progeny virions, in which only small amts. of the 7-kDa R peptide were found.
The 7-kDa R peptide was palmitoylated since it could be labeled
with [3H]palmitic acid, which explains its membrane assocn., slower
migration on gels, and high sensitivity in immunoblotting. The
present results are in contrast to previous findings showing equimolar amts. of R peptide and p15E in virions. The discrepancy, however,
can be explained by the presence of nonpalmitoylated R peptide in
virions, which were poorly detected by immunoblotting. A mechanistic
model is proposed. The uncleaved R peptide can, due to its lipid
modification, control the conformation of the ectodomain of the transmembrane protein and thereby govern membrane fusion.
132: 20884p Characteristics of aluminum biosorption by Sargassum fluitans biomass. Lee, Hak Sung; Volesky, Bohumil
(Department of Chemical Engineering, University of Ulsan, Ulsan
Nam, 680-749 S. Korea). Mar. Biotechnol. 1999, 1(4), 380-383 (Eng),
Springer-Verlag New York Inc.. A biomass of nonliving brown
seaweed Sargassum fluitans pretreated by different methods is capable
of taking up more than 10% (11 mEq/g) of its dry wt. in aluminum at
pH 4.5. There are indications that the biomass hydroxyl groups were
involved in sequestering the aluminum in the form of polynuclear
aluminum species. Aluminum-alginate complex (like cotton candy)
was formed in the aluminum sorption soln. as alginate was partially
released from the biomass. Aluminum uptake of S. fluitans biomass
was independent of residual alginate content in the biomass. Sodium
ion added for pH adjustment was not adsorbed at all in the presence
of aluminum ion.
132: 20890n Localization of periplasmic redox proteins of Alcaligenes faecalis by a modified general method for fractionating Gram-negative bacteria. Zhu, Zhenyu; Sun, Dapeng; Davidson, Victor L. (Department of Biochemistry, The University of
Mississippi Medical Center, Jackson, MS 39216-4505 USA). J. Bacteriol. 1999, 181(20), 6540-6542 (Eng), American Society for Microbiology. A lysozyme-osmotic shock method is described for fractionation of Alcaligenes faecalis which uses glucose to adjust osmotic
strength and multiple osmotic shocks. During phenylethylaminedependent growth, arom. amine dehydrogenase, azurin, and a single
cytochrome c were localized in the periplasm. Their induction patterns are different from those for the related quinoprotein methylamine dehydrogenase and its assocd. redox proteins.
132: 20891p Occurrence of free D-amino acids and aspartate
racemases in hyperthermophilic Archaea. Matsumoto, Megumi;
Homma, Hiroshi; Long, Zhiqun; Imai, Kazuhiro; Iida, Toshii; Maruyama, Tadashi; Aikawa, Yuko; Endo, Isao; Yohda, Masafumi (Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo,
Japan 113-0033). J. Bacteriol. 1999, 181(20), 6560-6563 (Eng),
American Society for Microbiology. The occurrence of free D-amino
acids and aspartate racemases in several hyperthermophilic archaea
was investigated. Aspartic acid in all the hyperthermophilic archaea

was highly racemized. The ratio of D-aspartic acid to total aspartic
acid was in the range of 43.0 to 49.1%. The crude exts. of the hyperthermophiles exhibited aspartate racemase activity at 70C, and aspartate racemase homologous genes in them were identified by PCR.
D-Enantiomers of other amino acids (alanine, leucine, phenylalanine, and lysine) in Thermococcus strains were also detected. Some
of them might be byproducts of aspartate racemase. It is proven that
D-amino acids are produced in some hyperthermophilic archaea,
although their function is unknown.
132: 20898w Enhancement of the lytic enzyme activity in
rifampicin-resistant mutants of Streptomyces recifensis var.
lyticus. Zhernosekova, I. V.; Kilochek, T. P.; Babenko, Yu. S.; Vinnikov, A. I. (Dnepropetr. Gos. Univ., Ukraine). Mikrobiol. Zh. 1999,
61(2), 15-24 (Russ), Institut Mikrobiologii i Virusologii NAN Ukraini.
Population variability of the producer of lytic enzymes Streptomyces
recifensis var. lyticus was studied at different stages of selection. It
has been established that the decrease of activity of strain II-29
already obtained by means of protoplasting was connected both with
the accumulation of inactive variants in the population and with the
changes in regulatory mechanisms of synthesis of extracellular
enzymes evidenced by the decrease of the strain sensitivity to catabolic
repression. High efficiency of rifampicin for selection of highly active
mutants have been shown. The strain 2P-15 has been obtained on
the basis of two stages of selection of rifampicin-resistant mutants.
The strain lytic activity reached 6300 U/mL that was twice as high
as the activity of the initial strain II-29.
132: 20899x A sampling of the yeast proteome. Futcher, B.;
Latter, G. I.; Monardo, P.; McLaughlin, C. S.; Garrels, J. I. (Cold
Spring Harbor Laboratory, Cold Spring Harbor, NY 11724 USA). Mol.
Cell. Biol. 1999, 19(11), 7357-7368 (Eng), American Society for Microbiology. In this study, we examd. yeast proteins by twodimensional (2D) gel electrophoresis and gathered quant. information
from about 1,400 spots. We found that there is an enormous range
of protein abundance and, for identified spots, a good correlation
between protein abundance, mRNA abundance, and codon bias. For
each mol. of well-translated mRNA, there were about 4,000 mols. of
protein. The relative abundance of proteins was measured in glucose
and ethanol media. Protein turnover was examd. and found to be
insignificant for abundant proteins. Some phosphoproteins were
identified. The behavior of proteins in differential centrifugation expts.
was examd. Such expts. with 2D gels can give a global view of the
yeast proteome.
132: 20903u The structure of the repeating unit of the O-specific polysaccharide from Marinomonas communis ATCC
27118(T). Zubkov, V. A.; Nazarenko, E. L.; Ivanova, E. P.; Gorshkova, N. M.; Gorshkova, R. P. (Pacific Institute of Bioorganic
Chemistry, Far East Division, Russian Academy of Sciences, Vladivostok, Russia 690022). Bioorg. Khim. 1999, 25(4), 290-292 (Russ),
MAIK Nauka. The O-specific polysaccharide from Marinomonas communis ATCC 27118(T) was isolated and characterized. Based on
carbohydrate anal., methylation, and 13C NMR spectroscopy, this
polysaccharide was found to be a homopolymer built of trisaccharide
repeating units.
132: 20906x Factors affecting the extreme acid resistance of
Escherichia coli O157:H7. Diez-Gonzalez, F.; Russell, J. B. (Section of Microbiology, Cornell University, Ithaca, NY 14833 USA). Food
Microbiol. 1999, 16(4), 367-374 (Eng), Academic Press. When
stationary phase Escherichia coli O157:H7 cells were subjected to
extreme acid shock (pH 20, 6 h, 37C) cell survival was as great as
10%, but culture conditions greatly affected the acid resistance.
Anaerobic cultures were more resistant to extreme acid shock if the
glucose concn. of the growth medium was high, acids accumulated,
and pH declined. By varying pH and acetate concn., it was possible
to demonstrate a high correlation (R2)086) between undissociated
acetate and extreme acid resistance. Because dissocd. acetate and
extreme acid resistance were poorly correlated (R2<001), it appeared
that the pH effects were being mediated via acetate dissocn. Propionate and butyrate were as effective as acetate, but formate, lactate,
benzoate and the uncoupler, carbonylcyanide m -chlorophenylhydrazone (CCCP), were much less effective in promoting extreme acidresistance. Acetate, propionate, butyrate, benzoate and CCCP all
decreased the intracellular pH of E. coli O157:H7, but the correlation
Issue 3, 2000
Page 3
between intracellular pH and extreme acid resistance was low (R2<001).

Cultures grown aerobically only needed half as much acetate to induce
extreme acid resistance as those grown anaerobically, and the addn.
of the reducing agent, cysteine, to anaerobic media made the stationary phase cells less responsive to acetate. An rpoS mutant of E. coli
O157:H7 was at least 100-fold more sensitive to acid shock than the
wild-type, and large amts. of acetate were needed to promote even a
small increase in viability. (c) 1999 Academic Press.
132: 20910u Effect of Saccharomyces cerevisiae ATCC 97 631
on aflatoxins production. Martins, H. M.; Bernardo, F. M.;
Martins, M. L. (Lab. Nacional Investigacao Veterinaria, Lisbon, Port.).
World Congr. Foodborne Infect. Intox., 4th 1998, 2, 1156-1160 (Eng).
Edited by Noeckler, Karsten. Federal Institute for Health Protection
of Consumers and Veterinary Medicine: Berlin, Germany. Aflatoxins
B1, B2, G1 and G2 are secondary metabolites produced by some
genetic competent strains of Aspergillus flavus, A. parasiticus and A.
nomius. The natural occurrence of these mycotoxins in food or feedstuffs can cause sever damage to human and animal health. Many
efforts have been made to control aflatoxin prodn. in foods. Modeling
natural competitive exclusion as a method for controlling aflatoxin
biosynthesis has been exptl. tried, but with different results. The
aim of the present study is to test the in vitro effect of Saccharomyces
cerevisiae ATCC 97 631 on aflatoxin biosynthesis when cultured
together with A. parasiticus (ATCC 15 517) in Modified CzapeckDox Broth (2% saccharose) (MCDB) and in cracked corn (CC) during
8 and 12 days at 28C. Quantification of aflatoxins B1, B2, G1 and
G2 was performed by HPLC and confirmed by TLC. Biomasses were
also detd. in MCDB cultures. Prodn. of aflatoxins (n)33) at 8 and
12 days with A. parasiticus was 0.978 (+ 0.077) and 1.152 (+0.115)
mg/50 mL, resp., in MCDB (P< 0.01) and 1.327 (+ 0.022) and 2.147
(+0.190) mg/50, resp., in CC (P<0.04). Cultures with S. cerevisiae
produced a very significant increase in the aflatoxin prodn. level: +
125% in MCDB at day 8 of incubation and + 134% at day 12. In
cracked corn productivity increases were higher: +194% and + 263%
at 8 and 12 days of incubation, resp. Biomasses of mixed cultures
were also increased (+ 11% and +12 % resp.). In terms of practical
application it is clear that S. cerevisiae is not an agent effective for
aflatoxins control; probably because it produce metabolites that are
intermediates in the biosynthesis pathway, thus increasing biosynthetic efficiency. Nevertheless, in lab. terms, it's an interesting finding, considering that S. cerevisiae can be used as a potential promoter
of in vitro productivity of aflatoxins.
132: 20913x ade9 is an allele of SER1 and plays an indirect
role in purine biosynthesis. Buc, Petra S.; Rolfes, Ronda J.
(Department of Biology, Georgetown University, Washington, DC
20057-1229 USA). Yeast 1999, 15(13), 1347-1355 (Eng), John Wiley
& Sons Ltd.. In this study we demonstrate that ade9 plays an indirect
role in purine biosynthesis as a non-functional allele of SER1 in
Saccharomyces cerevisiae. The SER1 locus, encoding 3-phosphoserine aminotransferase required for serine biosynthesis, is located
on chromosome XV and resides approx. where ade9 had previously
been mapped genetically. A minimal functional construct of SER1 is
necessary and sufficient to complement both the adenine- and serinerequiring phenotypes of ade9 strains. In addn., adequate exogenous
serine levels mask the adenine phenotype of ade9. A disruption of
SER1 behaves in the same manner phenotypically as the original
ade9 strain. Therefore, ade9 can be more accurately described as an
allele of SER1.
132: 20917b Induction of L-phenylalanine ammonia-lyase
and suppression of veratryl alcohol biosynthesis by exogenously added L-phenylalanine in a white-rot fungus Phanerochaete chrysosporium. Hattori, T.; Nishiyama, A.; Shimada,
M. (Wood Research Institute, Kyoto University, Uji, Kyoto, Japan).
FEMS Microbiol. Lett. 1999, 179(2), 305-309 (Eng), Elsevier Science
B.V.. The effects of exogenously added L-phenylalanine (L-Phe) on
the activities of L-phenylalanine ammonia-lyase (PAL) and 3,4dimethoxybenzyl alc. (veratryl alc., VA) biosynthesis in ligninolytic
cultures of Phanerochaete chrysosporium were investigated. Increasing PAL activity was detected in low nitrogen (LN) culture but not in
high nitrogen (HN) culture. The addn. of L-Phe into the LN culture
caused a 25-fold increase in enzyme activity, which clearly shows
that L-Phe, a substrate of the enzyme, served as an inducer of PAL.
The increase in activity of PAL triggered by nitrogen starvation was
Page 4
correlated with biosynthesis of VA. However, PAL induced by the

added L-Phe did not promote VA biosynthesis but suppressed the
biosynthesis probably due to NH4+ released from L-Phe.
132: 20918c Biosorption of copper and zinc by Cymodocea
nodosa. Sanchez, A.; Ballester, A.; Blazquez, M. L.; Gonzalez, F.;
Munoz, J.; Hammaini, A. (Facultad de Ciencias Quimicas, Departamento de Ciencia de los Materiales e Ingenieria Metalurgica, Universidad Complutense, Madrid, Spain). FEMS Microbiol. Rev. 1999,
23(5), 527-536 (Eng), Elsevier Science B.V.. The adsorption of the
two metal ions Cu and Zn in a single-component system by Cymodocea nodosa, a brown alga, under different pH conditions was investigated. The soln. pH significantly affected the exhibited uptake, being
max. at a pH value of 4.5. Multi-component mixt. biosorption in aq.
solns. is also reported. A comparison was made between the singlecomponent satn. uptake and the multi-component uptakes. To evaluate the two-metal sorption system performance, simple isotherm
curves had to be replaced by three-dimensional sorption isotherm
surfaces. In order to describe the isotherm surfaces math., three
Langmuir-type models were evaluated. The isotherms indicate a
competitive uptake with Cu being preferentially adsorbed. In addn.,
different tests were carried out to compare the process efficiency working continuously in small columns.
132: 20919d Bacterial respiration of arsenic and selenium.
Stolz, J. F.; Oremland, R. S. (Department of Biological Sciences,
Duquesne University, Pittsburgh, PA USA). FEMS Microbiol. Rev.
1999, 23(5), 615-627 (Eng), Elsevier Science B.V.. Oxyanions of
arsenic and selenium can be used in microbial anaerobic respiration
as terminal electron acceptors. The detection of arsenate and selenate respiring bacteria in numerous pristine and contaminated
environments and their rapid appearance in enrichment culture suggest that they are widespread and metabolically active in nature.
Although the bacterial species that have been isolated and characterized are still few in no., they are scattered throughout the bacterial
domain and include Gram-pos. bacteria, beta, gamma and epsilon
Proteobacteria and the sole member of a deeply branching lineage of
the bacteria, Chrysiogenes arsenatis. The oxidn. of a no. of org.
substrates (i.e. acetate, lactate, pyruvate, glycerol, ethanol) or hydrogen can be coupled to the redn. of arsenate and selenate, but the
actual donor used varies from species to species. Both periplasmic
and membrane-assocd. arsenate and selenate reductases have been
characterized. Although the no. of subunits and mol. masses differs,
they all contain molybdenum. The extent of the environmental impact
on the transformation and mobilization of arsenic and selenium by
microbial dissimilatory processes is only now being fully appreciated.
132: 20920x Characterization of dioxane and tetrahydrofuran degrading microorganisms and isolation of responsible
plasmid from a locally isolated Pseudomonas sp. Radwan, H.
H.; Yassin, M. A. (Faculty of Pharmacy, Helwan University, Egypt).
Al-Azhar J. Pharm. Sci. 1998, 22, 72-81 (Eng), Al-Azhar University, Faculty of Pharmacy. By an enrichment technique, some 140
locally isolated strains were tested for their ability to grow on aliph.
cyclic ethers [dioxane and THF (THF)]. Only 28 isolates utilized
dioxane and/or THF as a sole carbon and energy source. The most
potent candidates were Pseudomonas and Rhodococcus sp. A Pseudomonas sp. that degraded dioxane instead of or in combination with THF
was further characterized. The bacterium was able to degrade both
substrates, and neither substrate affected the utilization of the other.
A plasmid, with a mol. wt. of 61 kb, that seemed to carry genes
responsible for biodegrdn. of aliph. cyclic ethers in the Pseudomonas
sp. was isolated. Five cloned mutants of the Pseudomonas sp. were
obtained after plasmid curing; the mutants had lost the ability to
degrade both dioxane or THF.
132: 20923a Bacterial degradation of hydrocarbons as evidenced by respirometric analysis. Saadoun, I.; Al-Akhras,
M-Ali; Abu-Ashour, J. (Department of Biological Sciences, Jordan
University of Science and Technology, Irbid, Jordan 22110). Microbios 1999, 100(395), 19-25 (Eng), Faculty Press. The microbial
biodegradability of mineral oil and other hydrocarbons, namely hexane, decane and tetradecane was detd. using the Warburg const. vol.
respirometer. Results of oxygen uptake indicated that hexane and
tetradecane were more degradable than mineral oil and decane.
Rhodococcus erythropolis and Erwinia cancerogena showed the high-
Issue 3, 2000
est (0.866) and lowest (0.115) oxygen quotient (Qo2) values, resp.,
when exposed to mineral oil. Staphylococcus warneri and Enterobacter cloacae showed the highest (2.895) and (2.816) Qo2 values,
resp., when exposed to hexane; whereas E. cloacae and E. cancerogena showed the lowest Qo2 values (1.289 and 1.824), resp. Both R.
erythropolis and E. cloacae had the highest Qo2 values (2.859 and
2.289), resp., when exposed to tetradecane. More oxygen was consumed by R. erythropolis than the other bacterial cultures when
exposed to all hydrocarbons. In contrast, less oxygen was taken by
E. cancerogena than the other bacterial cultures when exposed to all
hydrocarbons, except for hexane.
132: 20925c Purification of PII and PII-UMP and in vitro studies of regulation of glutamine synthetase in Rhodospirillum
rubrum. Johansson, Magnus; Nordlund, Stefan (Department of
Biochemistry, Arrhenius Laboratories for Natural Sciences, Stockholm University, S-106 91 Stockholm, Swed.). J. Bacteriol. 1999,
181(20), 6524-6529 (Eng), American Society for Microbiology. The
PII protein from Rhodospirillum rubrum was fused with a histidine
tag, overexpressed in Escherichia coli, and purified by Ni2+-chelating
chromatog. The uridylylated form of the PII protein could be generated in E. coli. The effects on the regulation of glutamine synthetase
by PII, PII-UMP, glutamine, and R-ketoglutarate were studied in
exts. from R. rubrum grown under different conditions. PII and
glutamine were shown to stimulate the ATP-dependent inactivation
(adenylylation) of glutamine synthetase, which could be totally inhibited by R-ketoglutarate. Deadenylylation (activation) of glutamine
synthetase required phosphate, but none of the effectors studied had
any major effect, which is different from their role in the E. coli
system. In addn., deadenylylation was found to be much slower than
adenylylation under the conditions investigated.
132: 20927e The oxidative deamination of some amino acids
by metabolism products of Trichoderma. Smirnova, I. P.; Alekseev, S. B. (Russia's University for People's Friendship, Moscow,
Russia 117198). Biotekhnologiya 1998, (2), 68-72 (Russ), Biotekhnologicheskaya Akademiya RF. It has been established that the oxidative deamination of L-lysine, L-phenylalanine and L-methionine was
a function of the contents of these amino acids and of the acidity of
the medium. L-Lysine was the object of the most extensive degrdn.
The optimum pH value obsd. for L-lysine oxidative deamination was
5.8-6.0, for L-phenylamine 4.8-5.0 and for L-methionine 5.6-5.8
under the concns. of the amino acids in the medium 10 mM, 20 mM,
and 80 mM, resp.
132: 20928f The utilization of thiodiglycol by a bacterial
culture of the genus Pseudomonas. Medvedeva, N. G.; Zaitseva,
T. B.; Poliak, Yu. M.; Gridneva, Yu. A. (The Scientific Research
Center for Environmental Safety, Russ. Acad. Sci., St.-Petersburg,
Russia 197042). Biotekhnologiya 1998, (2), 89-92 (Russ), Biotekhnologicheskaya Akademiya RF. It was shown that using the technique
of culture enrichment a bacterial culture has been isolated from a soil
sample that was capable of using thiodiglycol (TDG) as a carbon
source and was characterized by a high resistance towards this
substrate. The vital activity of the culture still occurred at the concn.
of TDG in the medium as high as 20 g/l. The bacteria was affiliated
to the genus Pseudomonas. Mono- and dicarboxylic acids, 2-hydroxyethylthioglycolic and thiodiglycolic, were shown to be the intermediates of the initial steps of thiodiglycol metab.
132: 20930a -Poly(L-malate) production by Physarum
polycephalum: 13C nuclear magnetic resonance studies. Lee,
B.-S.; Maurer, T.; Kalbitzer, H. R.; Holler, E. (Institut fur Biophysik und physikalische Biochemie der Universitat Regensburg,
D-93040 Regensburg, Germany). Appl. Microbiol. Biotechnol. 1999,
52(3), 415-420 (Eng), Springer-Verlag. -Poly(L-malate) (PMLA)
prodn. in Physarum polycephalum has been followed by using D-[113
C]glucose and Ca13CO3. NMR studies of PMLA showed that the
13C label from [1-13C]glucose was incorporated in the presence of
CaCO3 into positions C-3(-CH2-) and C-4(-CO-) of the L-malate
repeating unit of PMLA. The 13C label from Ca13CO3 was incorporated
into position C-4 and indicated that not only the endogenous CO2
but also the exogenous CO2 from CaCO3 served significantly as a
carbon source for PMLA prodn. In the absence of CaCO3, the 13C
labeling pattern of PMLA from D-[1-13C]glucose was almost indistinguishable from that for the natural abundance 13C-NMR spectrum

of the polymer. These results indicated that L-malate used for PMLA
prodn. is synthesized either via carboxylation of pyruvate and redn.
of oxaloacetate in the presence of CaCO3 or via the oxidative tricarboxylic acid (TCA) cycle in the absence of CaCO3. Avidin strongly
inhibited the formation of L-malate via carboxylation; the 13C labeling pattern of PMLA in the presence of CaCO3 was almost identical
with that for the natural abundance spectrum when avidin was added,
indicating that L-malate utilized for PMLA prodn. was supplied under
this condition by the oxidative TCA cycle.
132: 20932c Metabolism of acetoin in Clostridium acetobutylicum ATCC 824. Wardwell, Stephanie Anne (Rice Univ., Houston,
TX USA). 1999. 202 pp. (Eng). Avail. UMI, Order No. DA9928612.
From Diss. Abstr. Int., B 1999, 60(5), 2004.
132: 20933d Mortierella strains as source of microbial surfaceactive molecules. Bandyopadhyay, S.; Chaudhuri, S.; Bhattacharyya, D. K. (Department Chemical Technology, Oil Technology
Division, Calcutta Univ., Calcutta, 700009 India). Fett/Lipid 1999,
101(11), 439-442 (Eng), Wiley-VCH Verlag GmbH. The prodn. of
surfactants from the fungal Mortierella strains were investigated
isolated from soil of a vegetable oil industry of Calcutta, India. The
surface-active property of the extd. fungal lipids and the amt. of
glycolipids and phospholipids in the lipids were detd. Three strains
of Mortierella were isolated: M. elongata IS-I, M. exigua IS-II, and
M. elongata IS-III. Biomass and total lipid contents of the fungi
were between 20-42 g/L and 20-43%, resp. The fatty acid compn.
of the samples showed the presence of linoleic and linolenic acid to a
considerable extent with a small amt. of arachidonic acid, eicosapentaenoic acid, and docosahexaenoic acid. Measurement of surface tension showed a redn. of surface tension of water by the addn. of 0.51% of fungal lipids. The fungal lipids and the solid portion recovered
from the aq. layer responded pos. to Molisch's test. The component
isolated from the solvent layer contained free fatty acids as detd. by
comparison with std. fatty acids on the TLC plate and by absorbance
obtained at 715 nm in a UV-visible spectrophotometer by the cupric
acetate method, revealing the presence of glycolipids. Column chromatog. sepn. of the lipids showed that neutral lipids formed 70-75%,
glycolipids 16-20%, and phospholipid 10-14% of the total lipids.
132: 20987z Fusarium avenacium, Peniophora, and their
fused forms with Phanerochaete chrysosporium for efficient
dioxin decomposition. Tachibana, Akiro; Ito, Kazutaka; Yamauchi, Yoko; Sasaki, Tetsuya; Suzuki, Kazuyoshi; Kondo, Takaaki
(Nippon Kokan Co., Ltd., Japan) Jpn. Kokai Tokkyo Koho JP
11 341,978 [99 341,978] (Cl. C12N1/15), 14 Dec 1999, JP Appl.
1998/105,884, 1 Apr 1998; 9 pp. (Japan). Provided are Fusarium
avenaceum (Corda:Fries) Saccardo strain 65 (65), Peniophora strain
267 (267), and their fused forms with Phanerochaete chrysosporium
(PC) for efficient decompn. of dioxins. A strain of Peniophora belonging to Corticiaceae is also used for the dioxin decompn. The rates of
decompn. of 2,7-Dichlorodibenzo-p-dioxin (2,7-DCDD) in a 30day study by strain 65, strain 267, strain PCx65, PCx267, and PC
were 82.9, 75.2, 99.1, 87.5, and 67.5%, resp.
132: 32955a Preliminary physicochemical evaluations and
bioactive properties of column fractions from the lichen. Esimone, C. O.; Adikwu, M. U.; Ebi, G. C.; Anaga, A.; Njoku, C.
(Faculty of Pharmaceutical Sciences University of Nigeria, Nsukka,
Nigeria). Boll. Chim. Farm. 1999, 138(4), 169-175 (Eng), Societa
Editoriale Farmaceutica. The chloroform-sol. portion of an ethanolic
ext. of the lichen Ramalina farinacea was sepd. by column chromatog.
using n-hexane: ethylacetate (4:1) as the mobile phase and silica gel
S as the stationary phase. Four column fractions (B, C, H and I) and
the n-hexane-sol. portion of the ethanolic ext. (N) were screened for
possible biol. activity by the brine shrimp lethality bioassay technique.
Some physicochem. evaluations of the column fractions such as pH
detns., UV absorption spectra, soly. profile and some chem. tests were
also carried out. Phytochem. and elemental anal. of the crude lichen
material was equally carried out. The results showed that all the
fractions and the n-hexane ext. were biol. active with a bioactivity
of N>B>C>I>H. Fractions B and C were both acidic and evaluation
of their soly. profile, UV spectra and some chem. reactions suggest
that the fractions are possibly depsides and depsidones, resp.
Issue 3, 2000
Page 5
132: 32957c Isolation of nitrogen-fixing bacteria containing

molybdenum-independent nitrogenases from natural environments. Loveless, Telisa M.; Saah, J. Royden; Bishop, Paul E.
(USDA Agricultural Research Service and Department of Microbiology, North Carolina State University, Raleigh, NC 27695-7615 USA).
Appl. Environ. Microbiol. 1999, 65(9), 4223-4226 (Eng), American
Society for Microbiology. Seven diazotrophs that grow well under
Mo-deficient, N2-fixing conditions were isolated from a variety of
environments. These isolates fall in the subdivision of the class
Proteobacteria and have genes that encode the Mo nitrogenase (nitrogenase 1) and the V nitrogenase (nitrogenase 2). Four of the isolates
also harbor genes that encode the Fe-only nitrogenase (nitrogenase
3).
132: 32959e Cysteine biosynthesis in Saccharomyces cerevisiae: a new outlook on pathway and regulation. Ono, BunIchiro; Hazu, Toshiya; Yoshida, Sayaka; Kawato, Takahiro; Shinoda, Sumio; Brzvwczy, Jerzy; Paszewski, Andrzej (Department of
Biotechnology, Faculty of Science and Engineering, Ritsumeikan
University, Kusatsu, Japan 525-8577). Yeast 1999, 15(13), 13651375 (Eng), John Wiley & Sons Ltd.. Using a Saccharomyces cerevisiae strain having the activities of serine O-acetyl-transferase
(SATase), O-acetylserine/O-acetylhomoserine sulfydrylase (OAS/OAH
SHLase), cystathionine -synthase (-CTSase) and cystathionine
-lyase (-CTLase), we individually disrupted CYS3 (coding for
-CTLase) and CYS4 (coding for -CTSase). The obtained gene
disruptants were cysteine-dependent and incorporated the radioactivity of 35S-sulfate into homocysteine but not into cysteine or glutathione. We concluded, therefore, that SATase and OAS/OAH
SHLase do not constitute a cysteine biosynthetic pathway and that
cysteine is synthesized exclusively through the pathway constituted
with -CTSase and -CTLase; note that OAS/OAH SHLase supplies
homocysteine to this pathway by acting as OAH SHLase. From
further investigation upon the cys3-disruptant, we obtained results
consistent with our earlier suggestion that cysteine and OAS play
central roles in the regulation of sulfate assimilation. In addn., we
found that sulfate transport activity was not induced at all in the
cys4-disruptant, suggesting that CYS4 plays a role in the regulation
of sulfate assimilation.
132: 32961z A mutant of Gluconobacter oxydans deficient in
gluconic acid dehydrogenase. Gupta, A.; Felder, M.; Verma, V.;
Cullum, J.; Qazi, G. N. (Division of Biotechnology, Regional Research
Laboratory, Jammu Tawi, India). FEMS Microbiol. Lett. 1999, 179(2),
501-506 (Eng), Elsevier Science B.V.. Gluconobacter oxydans ATCC
9937 was subjected to transposon mutagenesis using Tn5. A nonpigmented mutant was shown to be defective in gluconic acid dehydrogenase and to produce gluconic acid from glucose, whereas the parent
strain produced 2,5-diketogluconic acid. Cloning and sequencing of
the region contg. the Tn5 insertion showed that the insertion point
occurred in an open reading frame homologous (42% amino acid
identity) to the ribF genes of Pseudomonas fluorescens and Escherichia coli. The resulting lack of a riboflavin cofactor would explain the
loss of enzyme activity.
132: 32963b Alterations in macromolecular composition and
cell wall integrity by ciprofloxacin in Mycobacterium smegmatis. Verma, I.; Rohilla, A.; Khuller, G. K. (Dept. of Biochemistry,
Postgraduate Institute of Medical Education and Research, Chandigarh, India). Lett. Appl. Microbiol. 1999, 29(2), 113-117 (Eng), Blackwell Science Ltd.. The present study has been undertaken to explore
the biochem. mechanism of antimycobacterial action of a potent fluoroquinolone i.e. ciprofloxacin in Mycobacterium smegmatis. Cells grown
in the presence of a subinhibitory concn. (IC50) of ciprofloxacin had
a significantly lower content of all the major macromols. i.e DNA,
RNA, proteins and lipids with max. inhibition in DNA concn. as
compared to control. Significant quant. changes were also obsd. in
the various chem. constituents of cell wall of ciprofloxacin grown cells.
A decrease in the no. of binding sites for a fluorescent probe L-anilinonaphthalene-8-sulfonate (ANS) in ciprofloxacin grown cells suggested structural changes on the cell surface. Significant changes
were also obsd. in the morphol. of cells grown in the presence of
ciprofloxacin by SEM (SEM). Our results suggest that ciprofloxacin
exerts its antimycobacterial activity by affecting the cell wall as well
as various macromols., particularly DNA, the vital component for cell
survival and growth.
Page 6
132: 32965d Bacterial sources of hopanoids in recent sediments: improving our understanding of ancient hopane biomarkers. Farrimond, Paul; Fox, Paul A.; Innes, Helen E.; Miskin,
Ian P.; Head, Ian M. (Fossil Fuels and Environmental Geochemistry
(NRG), University of Newcastle upon Tyne, Newcastle upon Tyne,
UK NE1 7RU). Ancient Biomol. 1998, 2(2-3), 147-166 (Eng), Harwood Academic Publishers. Org. geochem. and mol. biol. have been
jointly applied in a study to det. the compn. of bacterially derived
hopanoids in the sediments of Priest Pot, a small lake in the English
Lake District, and to characterize the bacterial populations that source
the hopanoids within this lake. Anal. by GC/MS of acetylated aliquots of org. solvent-sol. org. matter in the sediments showed high
concns. of bacteriohopanetetrol. More highly functionalized hopanoids were detd. by GC/MS following treatment with periodic acid
and sodium borohydride which cleaves the polyfunctionalised side
chain to produce C32, C31 and C30 hopanol products representative of,
resp., tetra-, penta- and hexafunctionalized bacteriohopanoids
present in the original samples. These data indicate that composite
tetrafunctionalized bacteriohopanoids are present in far greater
abundance than bacteriohopanetetrol, the commonly presumed precursor of hopanoids in ancient samples. Penta- and hexafunctionalized
hopanoids are also more abundant. Thus, the biol. precursors of hopanoids in the geosphere are more diverse than previously thought.
The bacterial populations present in anoxic sediments from the lake
have been characterized by rRNA sequence-based analyses. These
revealed that the dominant bacterial populations present within the
sediments are not related to known hopanoid-synthesizing taxa. This
would suggest that it is principally the bacterial populations present
in the water column that source the hopanoids found in the sediments. This is consistent with current knowledge of the range of
bacteria known to produce hopanoids, since no obligately anaerobic
bacteria are known that synthesize these biomarkers. However, the
majority of rRNA sequences recovered belonged to novel evolutionary
lines of descent of which there are no known representatives among
cultured bacteria. Without cultures of the bacteria that these rRNA
sequences represent it is impossible to det. if these bacteria are capable
of synthesizing hopanoids.
132: 32967f Saccharothrix tangerinus sp. nov., the producer
of the new antibiotic formamicin: taxonomic studies. Kinoshita, Naoko; Igarashi, Masayuki; Ikeno, Souichi; Hori, Makoto;
Hamada, Masa (Institute of Microbial Chemistry, Tokyo, Japan 1410021). Actinomycetologica 1999, 13(1), 20-31 (Eng), Society for Actinomycetes Japan. The taxonomy of a soil isolate, strain MK2791F2 which produced a new antifungal antibiotics formamicin, was
studied. We propose that the strain should belong to a new species
of the genus Saccharothrix with the new name Saccharothrix tangerinus sp. nov., the type strain being MK27-91F2 () JCM 10302 ) IFO
16184 ) FERM P-16053). This new species was characterized by a
type III cell wall, galactose and rhamnose as whole-cell sugars, type
PII phospholipids, MK-9(H4) menaquinone, fatty acid components of
i-16:0, i-14:0, i-15:0, 16:0, 16:1, 17:1 and i-16:1, but lacking mycolic acids, a G+C content in DNA of 74 mol %, and a base sequence
of 16S rRNA gene.
132: 32969h Excretion products of algae and their occurrence in Solar Lake, Taba, Egypt. Badawy, M. I.; Abou-Waly,
H. F.; Ali, G. H. (Water Pollution Research Department, National
Research Center, Cairo, Egypt). Int. J. Environ. Health Res. 1999,
9(3), 233-243 (Eng), Carfax Publishing Ltd.. The present study
aimed to investigate the nature of org. compds. liable to be released
by blue-green algae (cyanobacteria) and org. compds. detected in the
Solar Lake in Taba, Egypt. The liq.-liq. extn. method and liq. chromatog. technique were applied for extn., clean-up and sepn. of the
orgs. Gas chromatog./mass spectrometry (GC/MS) was used to carry
out qual. anal. The most dominant species in the algal samples collected from Solar lake in Taba are Aphanotheca stagnina, Synechococcus aeruginosus, Oscillatoria limnetica and Microcoleus chthonoplastes. n-Alkane hydrocarbons that ranged from C12 to C25 were
identified in lake water samples, most of them were derived from
algal species. Several fatty acids were isolated from water samples,
with tridecanoic acid (C13), tetradecanoic acid (C14) and hexadecanoic
acid (C16) being prominent. Meanwhile, the identification of fatty
acids from algal suspension samples indicated the predominance of
hexadecanoic acid (C16) and heptadecanoic acid (C17), with carbon
nos. that ranged from C10 to C17. The dominant compds. were C15,
Issue 3, 2000
C17, C17:1, C19 n-alkanes. C13, C15, C16 fatty acids, alkyl esters of Pr
decanoate, esters of benzenedicarboxlate, di-Ph amine, geosmin and
2-Me isoborneol in water and in algal suspension samples were probably enough to distinguish the cyanobacterial mat community of a
saline lake shore.
132: 32972d The A-factor regulatory cascade leading to streptomycin biosynthesis in Streptomyces griseus: identification
of a target gene of the A-factor receptor. Ohnishi, Yasuo; Kameyama, Shogo; Onaka, Hiroyasu; Horinouchi, Sueharu (Department of Biotechnology, Graduate School of Agriculture and Life
Sciences, The University of Tokyo, Tokyo, Japan 113-8657). Mol.
Microbiol. 1999, 34(1), 102-111 (Eng), Blackwell Science Ltd.. In
Streptomyces griseus, A-factor (2-isocapryloyl-3R-hydroxymethyl-butyrolactone) at an extremely low concn. triggers streptomycin
biosynthesis and cell differentiation by binding a repressor-type receptor protein (ArpA) and dissocg. it from DNA. An A-factor-responsive
transcriptional activator (AdpA) able to bind the promoter of strR, a
pathway-specific regulatory gene responsible for transcription of other
streptomycin biosynthetic genes, was purified to homogeneity and
adpA was cloned by PCR on the basis of amino acid sequences of
purified AdpA. adpA encoding a 405-amino-acid protein contg. a
helix-turn-helix DNA-binding motif at the central region showed
sequence similarity to transcriptional regulators in the AraC/XylS
family. The -35 and -10 regions of the adpA promoter were a target
of ArpA; ArpA bound the promoter region in the absence of A-factor
and exogenous addn. of A-factor to the DNA-ArpA complex immediately released ArpA from the DNA. Consistent with this, S1
nuclease mapping showed that adpA was transcribed only in the presence of A-factor and strR was transcribed only in the presence of
intact adpA. Furthermore, adpA disruptants produced no streptomycin and overexpression of adpA caused the wild-type S. griseus strain
to produce streptomycin at an earlier growth stage in a larger amt.
On the basis of these findings, we propose here a model to demonstrate
how A-factor triggers streptomycin biosynthesis at a late exponential
growth stage.
132: 32973e Lanostanoid triterpenes from Ganoderma lucidum. Gonzalez, Antonio G.; Leon, Francisco; Rivera, Augusto; Munoz, Claudia M.; Bermejo, Jaime (Instituto Universitario de BioOrganica Antonio Gonzalez Instituto de Productos Naturales y
Agrobiologia, C.S.I.C., La Laguna, Spain 38206). J. Nat. Prod. 1999,
62(12), 1700-1701 (Eng), American Chemical Society. Two new
Me
Me
Me
Me
Me
HO
H
Me Me
I R=CO2 H
II R=CHO
lanostanoids, lucidadiol (I) and lucidal (II), were isolated from an

ethanolic ext. of Ganoderma lucidum, together with the known compds. ganodermadiol; ganodermenonol; ganoderic acid DM, ergosterol,
22,23-dihydroergosterol; ergosta-7,22-dien-3-one; fungisterol; ergosta-4,6,8(14),22-tetraen-3-one; and ergosterol peroxide. The
structures of I and II were detd. based on spectral evidence.
132: 32974f Genetic and biochemical characterization of a
high-affinity betaine uptake system (BusA) in Lactococcus
lactis reveals a new functional organization within bacterial
ABC transporters. Obis, David; Guillot, Alain; Gripon, JeanClaude; Renault, Pierre; Bolotin, Alexander; Mistou, Michel-Yves
(Unite de Biochimie et Structure des Proteines, Institut National de
la Recherche Agronomique, 78352 Jouy-en-Josas, Fr.). J. Bacteriol.
1999, 181(20), 6238-6246 (Eng), American Society for Microbiology.
The cytoplasmic accumulation of exogenous betaine stimulates the
growth of Lactococcus lactis cultivated under hyperosmotic conditions. We report that L. lactis possesses a single betaine transport
system that belongs to the ATP-binding cassette (ABC) superfamily
of transporters. Through transposon mutagenesis, a mutant deficient
in betaine transport was isolated. We identified two genes, busAA
and busAB, grouped in an operon, busA (betaine uptake system).
The transcription of busA is strongly regulated by the external osmolality of the medium. The busAA gene codes for the ATP-binding
protein. busAB encodes a 573-residue polypeptide which presents

two striking features: (i) a fusion between the regions encoding the
transmembrane domain (TMD) and the substrate-binding domain
(SBD) and (ii) a swapping of the SBD subdomains when compared to
the Bacillus subtilis betaine-binding protein, OpuAC. BusA of L.
lactis displays a high affinity towards betaine (Km ) 1.7 M) and is
an osmosensor whose activity is tightly regulated by external osmolality, leading the betaine uptake capacity of L. lactis to be under
dual control at the biochem. and genetic levels. A protein presenting
the characteristics predicted for BusAB was detected in the membrane
fraction of L. lactis. The fusion between the TMD and the SBD is
the first example of a new organization within prokaryotic ABC
transporters.
132: 32976h Ferric enterochelin transport in Yersinia enterocolitica: molecular and evolutionary aspects. Schubert, S.; Fischer, D.; Heesemann, J. (Max von Pettenkofer-Institut, 80336
Munich, Germany). J. Bacteriol. 1999, 181(20), 6387-6395 (Eng),
American Society for Microbiology. Yersinia enterocolitica is well
equipped for siderophore piracy, encompassing the utilization of siderophores such as ferrioxamine, ferrichrome, and ferrienterochelin. In
this study, we report on the mol. and functional characterization of
the Yersinia fep-fes gene cluster orthologous to the Escherichia coli
ferrienterochelin transport genes (fepA, fepDGC, and fepB) and the
esterase gene fes. In vitro transcription-translation anal. identified
polypeptides of 30 and 35 kDa encoded by fepC and fes, resp. A
frameshift mutation within the fepA gene led to expression of a
truncated polypeptide of 40 kDa. The fepD, fepG, and fes genes of Y.
enterocolitica were shown to complement corresponding E. coli mutants. Insertional mutagenesis of fepD or fes genes abrogates enterochelin-supported growth of Y. enterocolitica on iron-chelated media.
In contrast to E. coli, the fep-fes gene cluster in Y. enterocolitica
consists solely of genes required for uptake and utilization of enterochelin (fep) and not of enterochelin synthesis genes such as entF. By
Southern hybridization, fepDGC and fes sequences could be detected
in Y. enterocolitica biotypes IB, IA, and II but not in biotype IV
strains, Yersinia pestis, and Yersinia pseudotuberculosis strains. According to sequence alignment data and the coherent structure of the
Yersinia fep-fes gene cluster, we suggest early genetic divergence of
ferrienterochelin uptake determinants among species of the family
Enterobacteriaceae.
132: 32978k Transmembrane segment (TMS) VIII of the Na+/
citrate transporter CitS requires downstream TMS IX for insertion in the Escherichia coli membrane. Van Geest, Marleen;
Lolkema, Juke S. (Department of Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, 9751 NN Haren, Neth.). J. Biol. Chem. 1999, 274(42), 2970529711 (Eng), American Society for Biochemistry and Molecular
Biology. The amino acid sequence of the sodium ion-dependent citrate transporter CitS of K. pneumoniae contains 12 hydrophobic
stretches that could form membrane-spanning segments. A previous
anal. of the membrane topol. in Escherichia coli using the PhoA gene
fusion technique indicated that only nine of these hydrophobic segments span the membrane, while three segments, Vb, VIII and IX,
were predicted to have a periplasmic location. A topol. study of
C-terminally truncated CitS mols. in dog pancreas microsomes
revealed that the protein traverses the endoplasmic reticulum membrane 11 times. In agreement with the PhoA fusion data, segment
Vb was predicted to have a periplasmic location, but, in contrast,
segments VIII and IX were found to be membrane-spanning. In the
present study, using site-directed Cys labeling, the topol. of segments VIII and IX in the full-length CitS protein was detd. in the
E. coli membrane. Engineered cysteine residues in the loop between
the two segments were accessible to a membrane-impermeable thiol
reagent exclusively from the cytoplasmic side of the membrane,
demonstrating that transmembrane segments (TMSs) VIII and IX
are both membrane-spanning. It follows that the folding of CitS in
the E. coli and endoplasmic reticulum membrane is the same. Cysteine accessibility studies of CitS-PhoA fusion mols. demonstrated
that in the E. coli membrane segment VIII is exported to the periplasm in the absence of the C-terminal CitS sequences, thus explaining why the PhoA fusions do not correctly predict the topol. An
engineered cysteine residue downstream of TMS VIII moved from a
periplasmic to a cytoplasmic location when the fusion protein contg.
TMSs I-VIII was extended with segment IX. Thus, downstream seg-
Issue 3, 2000
Page 7
ment IX is both essential and sufficient for the insertion of segment

VIII of CitS in the E. coli membrane.
132: 32982g Occurrence and expression of glutathione-Stransferase-encoding bphK genes in Burkholderia sp. strain
LB400 and other biphenyl-utilizing bacteria. Bartels, Frank;
Backhaus, Silke; Moore, Edward R. B.; Timmis, Kenneth N.; Hofer,
Bernd (Division of Microbiology, National Research Centre for Biotechnology (GBF), 38124 Braunschweig, Germany). Microbiology
(Reading, U. K.) 1999, 145(10), 2821-2834 (Eng), Society for General
Microbiology. The gene bphK of Burkholderia sp. strain LB400 has
previously been shown to be located within the bph locus, which specifies the degrdn. of biphenyl (BP) and chlorobiphenyls, and to encode
a glutathione S-transferase (GST) which accepts 1-chloro-2,4dinitrobenzene (CDNB) as substrate. The specific physiol. role of this
gene is not known. It is now shown that the gene is expressed in the
parental organism and that GST activity is induced more than 20fold by growth of the strain on BP relative to succinate when these
compds. serve as sole carbon source. Approx. the same induction
factor was obsd. for 2,3-dihydroxybiphenyl 1,2-dioxygenase activity,
which is encoded by the 5'-adjacent bphC gene. This suggests that
the expression of bphK is coregulated with the expression of genes
responsible for the catabolism of BP. A bphK probe detected only a
single copy of the gene in strain LB400. A spontaneous BP- mutant
of the organism neither gave a signal with the bphK probe nor showed
CDNB-accepting GST activity, suggesting that this activity is solely
encoded by bphK. Complementation of the mutant with a bph gene
cluster devoid of bphK restored the ability to grow on BP, indicating
that bphK is not essential for utilization of this carbon source. BphK
activity proved to be almost unaffected by up to 100-fold differences
in proton concn. or ionic strength. The enzyme showed a narrow
range with respect to a variety of widely used electrophilic GST
substrates, accepting only CDNB. A no. of established lab. strains as
well as novel isolates able to grow on BP as sole carbon and energy
source were examd. for BphK activity and the presence of a bphK
analog. CDNB assays, probe hybridizations and PCR showed that
several, but not all, BP degraders possess this type of GST activity
and/or a closely related gene. In all bacteria showing BphK activity,
this was induced by growth on BP as sole carbon source, although
activity levels differed by up to 10-fold after growth on BP and by
up to 60-fold after growth on succinate. This resulted in a variation
of induction factors between 2 and 30. In the majority of bphK+
bacteria examd., the gene appeared to be part of LB400-like bph
gene clusters. DNA sequencing revealed almost complete identity of
bphK genes from five different bph gene clusters. These results suggest that bphK genes, although not essential, fulfil a strain-specific
function related to the utilization of BPs by their host organisms.
The usefulness of BphK as a reporter enzyme for monitoring the
expression of catabolic pathways is discussed.
132: 32990h Siderophore production of Vibrio parahaemolyticus strains from different sources. Yamamoto, Shigeo; Okujo,
Noriyuki; Miyoshi, Shin-Ichi; Shinoda, Sumio; Narimatsu, Shizuo
(Faculty of Pharmaceutical Sciences, Okayama University, Okayama,
Japan 700-8530). Microbiol. Immunol. 1999, 43(9), 909-912 (Eng),
Center for Academic Publications Japan. Vibrio parahaemolyticus
strains isolated from different sources were assayed for their ability
to produce a siderophore, vibrioferrin, under iron-limited growth
conditions. The mean value, std. error of mean (M vibrioferrin in
spent culture supernatant/optical d. at 660 nm) was 832.3, 66.9 for
clin. isolates (n 44), which was significantly higher (P 0.01) than
those for food isolates (461.0, 66.5; n 37) and coastal isolates (378.8,
37.2; n 26). This suggests that greater productivity of vibrioferrin by
clin. isolates may be assocd. with a selective advantage for survival
and proliferation under conditions of iron-limitation such as in the
intestine.
132: 33002z Cultural study and illudin S production of medicinal mushroom Omphalotus olearius (DC.:Fr.) Fay. (Agaricales s.l.) from Israel. Weis, Alexander L.; Solomko, Elvira F.;
Buchalo, Asya S.; Wasser, Solomon P.; Mitropolskaya, Nadezhda
Yu.; Grigansky, Andrey Ph.; Gorovits, Elena L. (International Myko
Biologics, Inc., San Antonio, TX 78245 USA). Int. J. Med. Mushrooms
1999, 1(1), 93-103 (Eng), Begell House, Inc.. The morphol. and
cultural characteristics of vegetative mycelium of 11 investigated
strains of Omphalotus olearius (DC.:Fr.) Fay. (Omphalotaceae, Agari-
Page 8
cales s.l.) were studied. The hyphae were found to have numerous
clamps and formations of arthroconidia and crystals. The growth
rate of mycelium (to 7.5 mm/day) on the surface varied depending on
media compn. Prodn. of orange-red pigment and the high activity
of gelatinase, urease, cellulase, and -glucosidase was obsd. in pure
cultures. Optimum temp. was developed for growth appeared to be
28C, with an initial pH of complex media between 4.0 and 4.5. A
methodical scheme for obtaining std. liq. inoculum useful for physiol.
and biochem. investigations of O. olearius. Illudin S content in liq.
broth under submerged cultivation for all strains was detd. Depending on the strain, the content of illudin S was between 0.81 and 3.1
mg/mL. Strains 162, 167 and 169 have been selected for optimization
of illudin S prodn.
132: 33003a New insights into the pyrimidine salvage pathway of Saccharomyces cerevisiae: requirement of six genes
for cytidine metabolism. Kurtz, J. E.; Exinger, F.; Erbs, P.; Jund,
R. (Institut de Recherche contre les Cancers de l'Appareil Digestif,
UPR 9003 CNRS, Laboratoire de Genetique, Hopitaux Universitaires
de Strasbourg, F-67091 Strasbourg, Fr.). Curr. Genet. 1999, 36(3),
130-136 (Eng), Springer-Verlag. Cytidine metab. in the yeast Saccharomyces cerevisiae was analyzed by genetic and biochem. approaches. Disruption of a unique ORF (Genbank accession No.
U20865) bearing homol. with eucaryotic or bacterial cytidine deaminases abolished cytidine deaminase activity and resulted in 5-fluorocytidine resistance. The gene encoding cytidine deaminase will be
referred to as CDD1 (Genbank accession no. AF080089). The ability
to isolate mutants resistant to 5-fluorocytidine which mapped to five
other loci demonstrated the existence of a complex cytidine metabolic
network. Deciphering this network revealed several original features:(1) cytidine entry is mediated by the purine-cytosine transporter
(Fcy2p),(2) cytidine is cleaved into cytosine by the uridine nucleosidase (Urh1p),(3) cytidine is phosphorylated into CMP by the uridine
kinase (Urk1p),(4) a block in cytosine deaminase (Fcy1p), but not in
cytidine deaminase (Cdd1p), constitutes a limiting step in cytidine
utilization as a UMP precursor.
132: 33004b Inhibitory effects of gluconic acid on glucose
oxidation by Gluconobacter. Velizarov, S.; Beschkov, V.; Georgieva, T. (Institute of Chemical Engineering, Bulgarian Academy of
Sciences, 1113 Sofia, Bulg.). Dokl. Bulg. Akad. Nauk. 1997, 50(1112), 63-66 (Eng), Bulgarska Akademiya na Naukite. The inhibitory
effect of gluconic acid, a reaction product, on Gluconobacter was
studied and numerical values of Pcrit (max. inhibitor concn. above
which no growth is possible) and n (empirical const. generally called
toxic power) in the case of substrate-unlimited glucose oxidn. were
established. Pcrit and n values for product inhibition were 0.707 and
0.28, resp. The empirical const. n accounts for the fact that the product
concn. limit Pcrit can be reached in different ways: linearly (n)1),
slow initial drop in rate rate followed by a rapid approach to Pcrit
(n<1), or vice versa (n>1). The main inhibitory effect caused by
gluconic acid is the reduced pH value of the medium, resulting in a
slowing of the growth rate. As glucose oxidn. is growth assocd., this
leads to a proportional decease of the gluconic acid productivity of the
system. Using the Pcrit and n values reported here allows for prediction of product inhibition kinetics for a given substrate (i.e. glucose)
concn. in Gluconobacter cultures which do not become oxygen limited.
132: 33005c Anaerobic oxidation of thiosulfate to tetrathionate by obligately heterotrophic bacteria, belonging to the
Pseudomonas stutzeri group. Sorokin, D. Y. D. Y.; Teske, A.;
Robertson, L. A.; Kuenen, J. G. (Institute of Microbiology, Russian
Academy of Sciences, Moscow, Russia 117811). FEMS Microbiol. Ecol.
1999, 30(2), 113-123 (Eng), Elsevier Science B.V.. A no. of strains
of heterotrophic bacteria were isolated from various environments on
the basis of their potential to oxidize inorg. sulfur compds. to tetrathionate. The isolates were screened for the ability to oxidize thiosulfate under denitrifying conditions. Many of them could grow anaerobically with acetate and nitrate, and eight strains could oxidize
thiosulfate to tetrathionate under the same conditions. In batch
cultures with acetate as carbon and energy source, most active anaerobic thiosulfate oxidn. occurred with N2O as electron acceptor. The
level of anaerobic thiosulfate-oxidizing activity in cultures and cell
suspensions supplied with nitrate correlated with the activity of nitrite
reductase in cell suspensions. Some strains converted thiosulfate to
tetrathionate equally well with nitrite, nitrate and N2O as electron
Issue 3, 2000
acceptors. Others functioned best with N2O during anaerobic thiosulfate oxidn. The latter strains appeared to have a lower level of nitrite
reductase activity. Thiosulfate oxidn. under anaerobic conditions was
much slower than in the presence of oxygen, and was obviously
controlled by the availability of org. electron donor. The strains had
DNA-DNA similarity levels higher than 30%. Sequence anal. of the
16S rRNA gene of four selected isolates showed their affiliation to
specific genomovars of Pseudomonas stutzeri and the proposed new
species, Pseudomonas balearica. As shown by 16S rRNA sequence
anal. and DNA-DNA hybridization, the previously misnamed 'Flavobacterium lutescens' (ATCC 27951) is also a P. stutzeri strain which
can oxidize thiosulfate to tetrathionate aerobically and anaerobically
in the presence of N2O. The data suggest that tetrathionate-forming
heterotrophic bacteria, in particular those belonging to the P. stutzeri
'superspecies', can play a much more significant role in the biogeochem. cycles than was previously recognized.
132: 33006d Comparison of Nitrosospira strains isolated from
terrestrial environments. Jiang, Q. Q.; Bakken, L. R. (Department of Biotechnological Sciences, Agricultural University of Norway,
Aas, Norway). FEMS Microbiol. Ecol. 1999, 30(2), 171-186 (Eng),
Elsevier Science B.V.. Most of our knowledge about the physiol. of
ammonia-oxidizing bacteria is based on expts. with Nitrosomonas
europaea, which appears to be less ubiquitous than Nitrosospira. The
authors have isolated Nitrosospiras from widely different environments and compared their specific growth rate, substrate affinity,
urease activity, temp. response, pH tolerance and cell morphol. Two
of the strains had a variable morphol.; the spirals were less tightly
coiled than the classical Nitrosospira type and a fraction of the culture
had a vibrioid appearance. These vibrioid strains were also peculiar
in having a much higher apparent activation energy for ammonia
monooxygenase (AMO) (129 and 151 kJ mol-1) than that of the more
classical Nitrosospiras (78 and 79 kJ mol-1). The differences in morphol. and activation energy were congruent with the phylogeny of the
genes for 16S rRNA. The response to pH in the medium was
investigated for four strains. The oxidn. rate at the onset of the pH
exposure expt. was found to obey classical steady state enzyme kinetics, assuming that NH3 (not NH4+) is the rate-limiting substrate.
The calcd. half satn. consts. (Ks) for AMO were 6-11 M NH3. Growth
had a narrower pH range than oxidn. activity and appeared to be
restricted by pH-dependent factors other than NH3. All the isolated
strains were urease pos., with a specific urease activity ranging from
60 to 158% of their specific AMO activity. The urease activity was
unaffected by acetylene inhibition of the energy metab. The substrate
affinity for one strain was found to be around 670 M.
132: 33008f Microbial degradation of aromatic substances by
local bacterial isolates. III. Factors affecting degradation of
2,4-dichlorophenoxyacetic acid by Pseudomonas stutzeri. ElTayeb, O. M.; Megahed, S. A.; El-Azizi, M. (Microbiology Department and Biotechnology Center, Faculty of Pharmacy, Cairo University, Cairo, Egypt). Egypt. J. Biotechnol. 1998, 4, 84-90 (Eng),
Egyptian Society for Biotechnology. A strain of Pseudomonas stutzeri
(GH2) capable of utilizing the arom. compd. 2,4-dichlorophenoxyacetic acid (2,4-D) as the sole source of carbon was previously isolated.
In this report, using this isolate, factors affecting 2,4-D degrdn. as
well as the 2,4-D degrdn. parameters in shake flasks were studied.
Factors included the effect of the 2,4-D concn., initial biomass, aeration, agitation, temp. and pH. The role of the 2,4-D concn. was
found to be significant in increasing the length of the lag period
previously found to precede the degrdn. High concns. of 2,4-D
increased the lag period. Degrdn. with min. lag was obtained by
decreasing substrate concn. and increasing initial biomass at ambient
temp. and neutral pH. The reaction was not affected by agitation or
addn. of org. nutrients. The capacity for degrdn. was examd. by studying the effect of repeated addns. of 2,4-D. Repeated addns. of 2,4-D
after either complete or about 50% degrdn. of initial dose caused the
lag period to disappear and the apparent degrdn. rate became faster.
When a second addn. was made after complete disappearance of 2,4D, the same result was obtained even when the 2,4-D concn. was
doubled. The increase in the apparent rate of degrdn. appeared not
to be concn. dependent as evidenced by similar degrdn. slopes. This
observation points to a potential in the application of this strain for
bioscrubbing continuous quantities of industrial effluents which are
rich in toxic arom. pollutants. However, the limits of substrate concn.
Issue 3, 2000
Page 9
which may be applied, as well as the limiting factors, possibly

nutritional, which may eventually control the no. of addns. need to
be examd.
procedures were objectively analyzed by an image anal. system. This

paper describes the progressive alteration of Salmonella typhimurium physiol. under salinity and starvation conditions.
132: 33009g Transport of sulfonium compounds. Characterization of the S-adenosylmethionine and S-methylmethionine permeases from the yeast Saccharomyces cerevisiae. Rouillon, Astrid; Surdin-Kerjan, Yolande; Thomas, Dominique (Centre
de Genetique Moleculaire, CNRS, 91198 Gif-sur-Yvette, Fr.). J.
Biol. Chem. 1999, 274(40), 28096-28105 (Eng), American Society for
Biochemistry and Molecular Biology. We report here the characterization and the mol. anal. of the two high affinity permeases that mediate the transport of S-adenosylmethionine (AdoMet) and S-methylmethionine (SMM) across the plasma membrane of yeast cells. Mutant
cells unable to use AdoMet as a sulfur source were first isolated and
demonstrated to lack high affinity AdoMet transport capacities.
Functional complementation cloning allowed us to identify the corresponding gene (SAM3), which encodes an integral membrane protein
comprising 12 putative membrane spanning regions and belonging to
the amino acid permease family. Among amino acid permease members, the closest relative of Sam3p is encoded by the YLL061 open
reading frame. Disruption of YLL061 was shown to specifically
lead to cells unable to use SMM as a sulfur source. Accordingly,
transport assays demonstrated that YLL061 disruption mutation
impaired the high affinity SMM permease, and YLL061 was therefore renamed MMP1. Further study of sam3 and mmp1 mutant
cells showed that in addn. to high affinity permeases, both sulfonium
compds. are transported into yeast cells by low affinity transport
systems that appear to be carrier-facilitated diffusion.
132: 33013d Control of Azospirillum brasilense NifA activity

by PII: effect of replacing Tyr residues of the NifA N-terminal
domain on NifA activity. Arsene, F.; Kaminski, P. A.; Elmerich,
C. (Departement des Biotechnologies, Centre National de la Recherche
Scientifique URA D1300, Unite de Physiologie Cellulaire, Institut
Pasteur, 75724 Paris, Fr.). FEMS Microbiol. Lett. 1999, 179(2), 339343 (Eng), Elsevier Science B.V.. It was previously reported that the
N-terminal domain of Azospirillum brasilense NifA was a neg. regulator of the NifA activity and that the PII protein prevented this inhibition under nitrogen fixing conditions. Here, the authors show that a
mutation of a single Tyr residue at position 18 of the N-terminal
domain of NifA led to an active NifA protein that did not require PII
for activation under nitrogen fixation conditions.
132: 33010a Adaptive responses of Ralstonia eutropha to feast

and famine conditions analyzed by flow cytometry. Muller, S.;
Bley, T.; Babel, W. (Sachsisches Institut fur Angewandte Biotechnologie (SIAB) an der Universitat Leipzig, 04318 Leipzig, Germany).
J. Biotechnol. 1999, 75(2,3), 81-97 (Eng), Elsevier Science Ireland
Ltd.. Results obtained by flow cytometry allow conclusions to be
drawn about how the physiol. states of Ralstonia eutropha JMP134
are connected with survival strategies under distinct growth conditions. During both feast and famine conditions the cells were found
to proceed through sharply sepd. phases of life. Two sources of carbon
and energy, one poor (0.02% phenol) and one rich (0.2% pyruvate and
0.1% yeast ext.) were chosen to study the cellular responses. Despite
the major differences in carbon source, when growth stages of the
bacteria on the two substrates were characterised in batch growth,
only minor differences were found in the time course of the membrane
potential-related fluorescence intensity (MPRFI). This also applied
to the rRNA content and the size-correlated forward scatter (FSC)
signal of the cells, both of which increased to high levels during the
(early) exponential growth phase. On the rich medium, DNA synthesis
initially occurred in an uncoupled manner, then a high rate of poly-hydroxybutyrate (PHB) formation followed when nutrients began
to be limiting. Under famine conditions, the cellular responses were
much more complex. PHB was synthesized, then DNA synthesis occurred in a eukaryotic mode, to be succeeded by renewed PHB
synthesis. To obtain defined cell physiol. states, the chemostat
technique was used in addn. to batch expts. The results obtained
clearly indicated that key events in cell physiol., including initiation
of DNA replication and overflow metab., occurred in a hierarchically
ordered manner and were tightly correlated with changes in the
environmental conditions of the bacterial cells.
132: 33011b Physiological changes of Salmonella typhimurium cells under osmotic and starvation conditions by image
analysis. Caro, A.; Got, P.; Baleux, B. (UMR CNRS-Universite
Montpellier II No. 5556, Laboratoire Hydrobiologie Marine et Continentale, Universite de Montpellier II, F-34095 Montpellier, Fr.).
B.V.. The effects of starvation and salinity on the physiol. of Salmonella typhimurium were investigated in a microcosm study. The
physiol. changes were monitored by using fluorochrome dyes, such as
DAPI (4',6-diamidino-2-phenylindole) for evaluation of the genomic
content, CTC (5-cyano-2,3-ditolyl tetrazolium chloride) for respiratory activity and SYTO 9 and propidium iodide for cytoplasmic
membrane damages. The metabolic activity of the cellular population was assessed with the method of Kogure (direct viable count), to
enumerate the substrate-responsive cells. These different staining
132: 33014e Role of multiple efflux pumps in Escherichia coli

in indole expulsion. Kawamura-Sato, K.; Shibayama, K.; Horii,
T.; Iimuma, Y.; Arakawa, Y.; Ohta, M. (Department of Bacteriology, Nagoya University School of Medicine, Nagoya, Aichi, Japan).
B.V.. The Escherichia coli chromosome encodes several multidrug
transporters. Despite their protective function against antibacterial
agents, the specific physiol. actions of these transporters are not fully
understood. E. coli produces indole, a metabolite of tryptophan, under
physiol. conditions. Defined inactivation of the acrEF gene, the
product of which is known as an energy-dependent multiple drug
efflux pump, decreased indole excretion while reintroduction of the
acrEF gene restored it. A acrEF mutant accumulated more intracellular indole than the parent. This mutant was more susceptible to
the growth-inhibitory effect of indole than the parent. These results
indicate that the AcrEF system plays a significant role in indole efflux.
132: 33016g A study on metabolism, oxygen uptake and hydrogen peroxide production of energy yielding substrates by Mycoplasma canis. Abdel Megid, R. (Microbiology Department, Faculty
of Pharmacy, Cairo University, Cairo, Egypt). Egypt. J. Biomed. Sci.
1999, 3, 125-138 (Eng), Egyptian Society for Biotechnology. Oxygen
uptake during the metab. of various substrates by whole cells of Mycoplasms canis (M. canis) and H2O2 accumulation together with the
oxidn. of NADH and L-R-glycerophosphate (GP) by cells lysed with
Triton X-100 was detd. for eight isolates of M. canis from various
sites in infected cattle. Oxidn. of glycerol at low concns. (not glucose),
NADH and GP by M. canis isolates was accompanied by the accumulation of large quantities of H2O2. However, metab. of glucose and
glycerol was indicated by a redn. in the pH of the suspending medium
(saline and catalase). Lysed cells oxidised NADH and GP with the
prodn. of approx. 1.0 mol H2O2/mL O2 taken up, indicating presence
of a GP oxidase. The importance of H2O2 prodn. as a factor in the
pathogenicity of M. canis indicates that it might depend upon the
availability of glycerol in vivo.
132: 33017h Metabolic flux analysis of Escherichia coli deficient in the acetate production pathway and expressing the
Bacillus subtilis acetolactate synthase. Yang, Yea-Tyng; Aristidou, Aristos A.; San, Ka-Yiu; Bennett, George N. (Department of
Bioengineering and Chemical Engineering, Institute of Biosciences
and Bioengineering, Rice University, Houston, TX 77005-1892 USA).
Metab. Eng. 1999, 1(1), 26-34 (Eng), Academic Press. Several approaches to reduce acetate accumulation in Escherichia coli cultures
have recently been reported. This redn. subsequently led to a significant enhancement in recombinant protein prodn. In those studies,
metabolically engineered E. coli strains with reduced acetate synthesis
rates were constructed through the modification of glucose uptake
rate, the elimination of crit. enzymes that are involved in the acetate
formation pathways, and the redirection of carbon flux toward less
inhibitory byproducts. In particular, it has been shown that strains
carrying the Bacillus subtilis acetolactate synthase (ALS) gene not
only produce less acetate but also have a higher ATP yield. Metabolic
flux anal. of carbon flux distribution of the central metabolic pathways
and at the pyruvate branch point revealed that this strain has the
ability to channel excess pyruvate to the much less toxic compd.,
acetoin. The main focus of this study is the systematic anal. of the
Page 10
effects of small perturbations in the host's existing pathways on the

redistribution of carbon fluxes. Specifically, a mutant with deleted
acetate kinase (ACK) and acetyl phosphotransferase (PTA) was
constructed and studied. Results from the metabolic anal. of carbon
redistribution show the ackA-pta mutation will reduce acetate level
at the expense of the growth rate. In addn., in the ackA-pta deficient
strain a much higher lactate formation rate with simultaneously lower
formate and ethanol synthesis rates was found. Expression of the B.
subtilis ALS in ackA-pta mutants further reduces acetate levels while
cell d. similar to that of the parent strain is attained. (c) 1999
Academic Press.
132: 33018j Response of the central metabolism in Corynebacterium glutamicum to the use of an NADH-dependent
glutamate dehydrogenase. Marx, Achim; Eikmanns, Bernhard
J.; Sahm, Hermann; De Graaf, Albert A.; Eggeling, Lothar (Institut fur Biotechnologie, Forschungszentrum Julich GmbH, D-52425
Julich, Germany). Metab. Eng. 1999, 1(1), 35-48 (Eng), Academic
Press. The extensive use of 13C enrichments in precursor metabolites
for flux quantification does not rely on NADPH stoichiometries and
can therefore be used to quantify reducing power fluxes. As an application of this concept, the NADPH fluxes were quantified in an
L-lysine producer of Corynebacterium glutamicum grown into metabolic and isotopic steady state with [1-13C]glucose. In this case,
where the organism's NADPH-dependent glutamate dehydrogenase
consumes reducing power, the NADPH flux generated is 210% (molar
flux relative to glucose uptake rate) with its major part (72% of the
total) generated via the pentose phosphate pathway activity. An
isogenic strain in which the glutamate dehydrogenase of C. glutamicum was replaced by the NADH-dependent glutamate dehydrogenase
of Peptostreptococcus asaccharolyticus was made and the metabolite
fluxes were again estd. The major response to this local perturbation
is a drastically reduced NADPH generation of only 139%. Most of
the NADPH (62% of the total) is now generated via the tricarboxylic
acid cycle activity. This shows the extraordinary flexibility of the
central metab. and provides a picture of the global regulatory properties of the central metab. Furthermore, a detailed anal. of the fluxes
and exchange fluxes within the anaplerotic reactions is given. It is
hypothesized that these reactions might also serve to balance the
total reducing power budget as well as the energy budget within the
cell. (c) 1999 Academic Press.
132: 33019k Regulated expression of the nifM of Azotobacter
vinelandii in response to molybdenum and vanadium supplements in Burk's nitrogen-free growth medium. Lei, Shi; Pulakat, Lakshmidevi; Gavini, Narasaiah (Department of Biological
Sciences, Bowling Green State University, Bowling Green, OH 43403
USA). Biochem. Biophys. Res. Commun. 1999, 264(1), 186-190 (Eng),
Academic Press. Azotobacter is a diazotrophic bacterium that harbors
three genetically distinct nitrogenases referred to as nif, vnf, and anf
systems. The nifM is an accessory gene located in the nif gene cluster
and is transcriptionally regulated by the NifA. However, Azotobacter
mutants that lack NifA are known to synthesize functional NifM and
this accessory protein is known to be needed for the activity of nitrogenase-2 and nitrogenase-3. To det. how the transcription of nifM
is regulated when Azotobacter is grown under conditions in which
nitrogenase-2 or nitrogenase-3 is expressed, we generated an Azotobacter vinelandii strain that carries a nifM:lacZ-kanamycin resistance gene cassette in its chromosome. In this strain the nifM open
reading frame was disrupted by the presence of a lacZ-kanamycin
resistance gene cassette so that it could not produce active NifM.
Moreover, the lacZ gene was placed under the transcriptional control
elements of the nifM gene so that the lacZ expression could be used
as a marker to det. the extent of expression of the nifM gene under
different growth conditions. These results show that this strain was
unable to grow in Burk's nitrogen-free medium supplemented with
either molybdenum or vanadium or lacking both metals, suggesting
that in the absence of functional NifM none of the nitrogenases was
active. It was also found that the nifM expression was differentially
regulated when the A. vinelandii cells were grown under conditions
that activate nitrogenase-2 and nitrogenase-3, as detd. by liq. -galactosidase activity measurements. These results suggest that the
transcriptional activators, VnfA and AnfA, may regulate the nifM
expression. (c) 1999 Academic Press.
132: 33021e Mineralization of 14C-labelled highly-condensed
polycyclic aromatic hydrocarbons in soils by Pleurotus sp.
Issue 3, 2000
Florida. Martens, R.; Wolter, M.; Bahadir, M.; Zadrazil, F. (Bundesforschungsanstalt fur Landwirtschaft, Institut fur Agrarokologie,
D-38116 Braunschweig, Germany). Soil Biol. Biochem. 1999, 31(13),
1893-1899 (Eng), Elsevier Science Ltd.. This paper shows the
potential of the fungus Pleurotus sp. Florida to degrade several PAH
compds. that are difficult to mineralize in untreated soils. This
capability was most pronounced in sand samples when growth of the
fungus was supported by an addnl. straw amendment in the sand
and bioavailability was not reduced by an adsorption of the PAH
compds.
132: 33023g All (S) stereoconfiguration of 7,10-dihydroxy8(E)-octadecenoic acid from bioconversion of oleic acid by
Pseudomonas aeruginosa. Gardner, Harold W.; Hou, Ching T.
(Bioactive Agents Research, NCAUR, ARS, USDA, Peoria, IL 61604
USA). J. Am. Oil Chem. Soc. 1999, 76(10), 1151-1156 (Eng), AOCS
Press. A previously established method was utilized to det. the stereoconfiguration of 7,10-dihydroxy-8(E)-octadecenoic acid (DHOE)
from bioconversion of oleic acid by Pseudomonas aeruginosa NRRL
strain B-18602 (PR3). The method involved formation of the (-)menthoxycarbonyl (MCO) deriv. of the two hydroxyls, oxidative cleavage of the double bond, and gas chromatog. (GC) anal. of the two
methyl-esterified diastereomeric fragments, Me 2-MCO-decanoate
and di-Me 2-MCO-octanedioate. As described by previous workers, the 2(S)-MCO derivs. elute at earlier times by GC than the
2(R)-MCO derivs. By comparing the GC anal. of the 2-MCO derivs.
obtained from DHOE with that obtained from a partially racemized
sample, DHOE was detd. to be 7(S), 10(S)-dihydroxy-8(E)-octadecenoic acid.
132: 33024h The entomopathogenic fungus Metarhizium
anisopliae alters ambient pH, allowing extracellular protease
production and activity. St Leger, Raymond J.; Nelson, Judd O.;
Screen, Steven E. (Department of Entomology, University of Maryland, College Park, MD 20742-4454 USA). Microbiology (Reading,
U. K.) 1999, 145(10), 2691-2699 (Eng), Society for General Microbiology. Ambient pH regulates the expression of virulence genes of
Metarhizium anisopliae, but it was unknown if M. anisopliae can
regulate ambient pH. Mutants of M. anisopliae altered in prodn. of
oxalic acid were evaluated for the interrelationship of ambient pH,
buffering capacity added to media, growth, and generation of extracellular proteases and ammonia. Wild-type and acid-overproducing
mutants [Acid(+)] grew almost as well at pH 8 as at pH 6, but acidnon-producing [Acid(-)] mutants showed limited growth at pH 8,
indicating that acid prodn. is linked to the ability to grow at higher
pH. Prodn. of ammonia by M. anisopliae was strongly stimulated by
low levels of amino acids in the medium when cells were derepressed
for nitrogen and carbon. Likewise, although Aspergillus fumigatus
and Neurospora crassa produced some ammonia in minimal media,
addn. of low levels of amino acids enhanced prodn. Ammonia prodn.
by A. fumigatus, N. crassa and M. anisopliae increased the pH of the
medium and allowed prodn. of subtilisin proteases, whose activities
are obsd. only at basic pH. In contrast, protease prodn. by the acid(+) mutants of M. anisopliae was greatly reduced because of the
acidification of the medium. This suggests that alkalinization by ammonia prodn. is adaptive by facilitating the utilization of proteinaceous nutrients. Collectively, the data imply that ammonia may have
functions related to regulation of the microenvironment and that it
represents a previously unconsidered virulence factor in diverse fungi
with the potential to harm tissues and disturb the host's immune
system.
132: 33025j Involvement of glutathione in the regulation of
respiratory oscillation during a continuous culture of Saccharomyces cerevisiae. Murray, Douglas B.; Engelen, Frank; Lloyd,
David; Kuriyama, Hiroshi (Microbiology Group, Cardiff School of
Biosciences, Cardiff University, Cardiff, UK CF10 3TL). Microbiology (Reading, U. K.) 1999, 145(10), 2739-2745 (Eng), Society for
General Microbiology. Respiratory oscillation occurred during aerobic
continuous culture of Saccharomyces cerevisiae. During oscillation,
phase-related changes in NAD(P)H and GSH levels occur. Perturbation of oscillation and inhibition of respiration occurred when GSH
or GSSG was injected; however, there was a phase delay in perturbation in the case of an injection during high respiration. The perturbation phase delay was not apparent when a combination of DLbuthionine-(S,R)-sulfoximine, GSH and 5-nitro-2-furaldehyde was

injected. Perturbation by GSH injection caused the intracellular GSH
concn. to increase, the GSSG concn. to decrease and the cessation of
ethanol uptake. NAD(P)H during perturbation was inversely related
to dissolved oxygen. Perturbation by calcium pantothenate and pyridoxal-HCl caused a period of enhanced respiration before oscillation
returned. These results suggest that the NAD+/NADH redox is not
directly involved in oscillation control and regulation involves glutathione metab. Possible regulation points include alc. dehydrogenase inhibition and/or respiratory-chain inhibition.
132: 33027m Extracellular metal-binding activity of the
sulfate-reducing bacterium Desulfococcus multivorans. Bridge,
Toni A. M.; White, Chris; Gadd, Geoffrey M. (Department of Biological Sciences, University of Dundee, Dundee, UK DD1 4HN). Microbiology (Reading, U. K.) 1999, 145(10), 2987-2995 (Eng), Society for
General Microbiology. Polarog. was used to measure the copperbinding ability of culture filtrates from a range of sulfate-reducing
bacteria (SRB), including pure cultures and environmental isolates.
Of those tested, Desulfococcus multivorans was shown to have the
greatest copper-binding capacity and this organism was used for
further expts. Extracellular copper- and zinc-binding activities of
Dc. multivorans culture filtrates from batch cultures increased over
time and reached a max. after 10 d growth. The culture filtrate was
shown to bind copper reversibly and zinc irreversibly. Twelve-dayold Dc. multivorans culture filtrates were shown to have a copperbinding capacity of 3.64(0.33 mol ml-1 with a stability const., log10K,
of 5.68(0.64 (n)4). The metal-binding compd. was partially purified from culture growth media by dichloromethane extn. followed by
HPLC using an acetonitrile gradient.
132: 33028n Inhibitory analysis of the energy metabolism of
the extremely haloalkaliphilic homoacetic bacterium Natroniella acetigena. Pusheva, M. A.; Pitryuk, A. V.; Netrusov, A. I.
(Institute of Microbiology, Russian Academy of Sciences, Moscow, Russia 117811). Microbiology (Moscow) 1999, 68(5), 565-567 (Eng),
MAIK Nauka/Interperiodica Publishing. The effect of ionophores and
dicyclohexylcarbodiimide (DCCD) on growing cultures of the extremely
haloalkaliphilic homoacetic bacterium Natroniella acetigena was
studied. Growth and acetogenesis in media with lactate or ethanol
at optimal pH value 9.7 and a salinity of 1.57% were completely
inhibited by DCCD, a potent inhibitor of F1F0-ATPase. The protonophore tetrachlorosalicylanilide and the Na+/H+ antiporter monensin produced a similar effect: growth and acetogenesis stopped
instantly after the addn. of these compds. to an 18-h culture. N.
acetigena was less sensitive to amiloride, which is an inhibitor of Na+
transport. Inhibitory anal. showed that the energy metab. of N. acetigena depends on the generation of a transmembrane electrochem.
gradient of Na+ via Na+/H+ antiport. Proton cycling via oxidative
phosphorylation and F1F0-ATPase are necessary for maintaining pHhomeostasis.
132: 33029p Bioenergetics of acetogenesis in the extremely
alkaliphilic homoacetogenic bacteria Natroniella acetigena
and Natronoincola histidinovorans. Pusheva, M. A.; Pitryuk, A.
V.; Detkova, E. N.; Zavarzin, G. A. (Institute of Microbiology, Russian Academy of Sciences, Moscow, Russia 117811). Microbiology
(Moscow) 1999, 68(5), 568-573 (Eng), MAIK Nauka/Interperiodica
Publishing. The energy metab. of the extremely alkaliphilic homoacetogens Natroniella acetigena and Natronoincola histidinovorans is
based on the acetyl-CoA pathway involving CO dehydrogenase, whose
activity is manifested at alk. pH values and is independent of the
presence of NaCl and bicarbonate ions. Inhibitory anal. showed that
the oxidn. of lactate and ethanol by N. acetigena and the oxidn. of
histidine by Natr. histidinovorans are coupled to ATP synthesis involving the electron transport chain and an H+-ATPase of the F1F0 type.
Cell de-energization by protonophores led to optical changes in resting N. acetigena cells; these changes were related to plasmolysis and
subsequent cell lysis as a result of the elimination of the active osmotic
barrier. Despite its fermentative metab., Natr. histidinovorans proved
to be obligately dependent on the activity of ion pumps.
132: 33032j Confirmation of the aerobic production of trimethylstibine by Scopulariopsis brevicaulis. Andrewes, Paul;
Cullen, William R.; Polishchuk, Elena (Environmental Chemistry
Group, Department of Chemistry, University of British Columbia,
Vancouver, BC Can. V6T 1Z1). Appl. Organomet. Chem. 1999, 13(9),
Issue 3, 2000
Page 11
659-664 (Eng), John Wiley & Sons Ltd.. The filamentous fungus
Scopulariopsis brevicaulis produces volatile trimethylstibine, found in
the culture headspace, when grown in an antimony(III)-rich medium
under aerobic conditions. The trimethylstibine was purged from
cultures using a continuous flow of compressed air and trapped in a
U-shaped tube contg. Supelcoport SP 2100 at -78C. The trap
contents were detd. by using GC-ICP-MS methodol. Typically
between 60 and 500 pg of trimethylstibine was trapped during
sampling (12 h) from cultures contg. 1000 g Sb ml-1 as potassium
antimony tartrate. The total prodn. of trimethylstibine over 18 days
of growth was estd. at 10 ng. Trimethylarsine was produced in greater
quantities than trimethylstibine, even though no arsenic compds. were
added to the medium.
132: 33037q Ultraviolet and osmotic stresses induce and
regulate the synthesis of mycosporines in the cyanobacterium
Chlorogloeopsis PCC 6912. Portwich, Anne; Garcia-Pichel, Ferran (Max-Planck-Institut fur Marine Mikrobiologie, D-28359 Bremen, Germany). Arch. Microbiol. 1999, 172(4), 187-192 (Eng),
Springer-Verlag. The cyanobacterium Chlorogloeopsis PCC 6912 was
found to synthesize and accumulate two putative UV sunscreen compds. of the mycosporine (mycosporine-like amino acid; MAA) type:
mycosporine-glycine and shinorine. These MAAs were not constitutively present in the cells; their synthesis could be induced specifically either by exposure to UVB radiation (280-320 nm) or by osmotic
stress, but not by other stress factors such as heat or cold shock,
nutrient limitation, or photooxidative stress. A significant synergistic
enhancement of MAA synthesis was obsd. when both stress factors
were applied in combination. Although osmotic stress could induce
MAA synthesis, comparison of the intracellular contents of MAAs
with those of sugar osmolytes (glucose and trehalose) indicated that
MAAs play no significant role in attaining osmotic homeostasis. UVB
strongly enhanced the accumulation of shinorine, whereas osmotic
stress had a more pronounced effect on mycosporine-glycine. This
differential effect on the steady-state contents of each MAA could be
explained either by differential regulation of biosynthesis or by differential loss rates of MAAs (leakage) under each condition. A
preferential leakage of mycosporine-glycine from the cells after a
hypoosmotic shock was detected. The results are interpreted in terms
of an adaptive necessity for a combined regulatory control responding
to both UV and external osmotic conditions in organisms that accumulate water-sol. sunscreens intracellularly.
132: 33038r Phototrophic utilization of toluene under anoxic
conditions by a new strain of Blastochloris sulfoviridis. Zengler, Karsten; Heider, Johann; Rossello-Mora, Ramon; Widdel,
Friedrich (Max-Planck-Institut fur Marine Mikrobiologie, D-28359
Bremen, Germany). Arch. Microbiol. 1999, 172(4), 204-212 (Eng),
Springer-Verlag. The capacity of anoxygenic phototrophic bacteria
to utilize arom. hydrocarbons was investigated in enrichment cultures
with toluene. When mineral medium with toluene (provided in an
inert carrier phase) was inoculated with activated sludge and incubated under IR illumination (> 750 nm), a red-to-brownish culture
developed. Agar diln. series indicated the dominance of two types of
phototrophic bacteria. One type formed red colonies, had rod-shaped
cells with budding division, and grew on benzoate but not on toluene.
The other type formed yellow-to-brown colonies, had oval cells, and
utilized toluene and benzoate. One strain of the latter type, ToP1,
was studied in detail. Sequence anal. of the 16S rRNA gene and
DNA-DNA hybridization indicated an affiliation of strain ToP1 with
the species Blastochloris sulfoviridis, a member of the R-subclass of
Proteobacteria. However, the type strain (DSM 729) of Blc. sulfoviridis grew neither on toluene nor on benzoate. Light-dependent
consumption of toluene in the presence of carbon dioxide and formation of cell mass by strain ToP1 were demonstrated in quant. growth
expts. Strain ToP1 is the first phototrophic bacterium shown to utilize
an arom. hydrocarbon. In the supernatant of toluene-grown cultures
and in cell-free exts. incubated with toluene and fumarate, the formation of benzylsuccinate was detected. These findings indicate that
the phototrophic bacterium activates toluene anaerobically by the
same mechanism that has been reported for denitrifying and sulfatereducing bacteria. The natural abundance of phototrophic bacteria
with the capacity for toluene utilization was examd. in freshwater
habitats. Counting series revealed that up to around 1% (1.8 105
cells per g dry mass of sample) of the photoheterotrophic population
cultivable with acetate grew on toluene.
Page 12
132: 33041m Degradation of 2,4-dinitrophenol and selected

nitroaromatic compounds by Sphingomonas sp. UG30. Zablotowicz, R. M.; Leung, K. T.; Alber, T.; Cassidy, M. B.; Trevors, J.
T.; Lee, H.; Veldhuis, L.; Hall, J. C. (Southern Weed Science
Research Unit, United States Department of Agriculture - Agricultural Research Service, Stoneville, MS 38776 USA). Can. J. Microbiol. 1999, 45(10), 840-848 (Eng), National Research Council of
Canada. Sphingomonas strain UG30 mineralizes both p-nitrophenol
(PNP) and pentachlorophenol (PCP). Our current studies showed
that UG30 oxidatively metabolized certain other p-substituted nitrophenols, i.e., p-nitrocatechol, 2,4-dinitrophenol (2,4-DNP), and 4,6dinitrocresol with liberation of nitrite. 2,6-DNP, o- or m-nitrophenol, picric acid, or the herbicide dinoseb were not metabolized. Studies
using 14C-labeled 2,4-DNP indicated that in glucose-glutamate
broth cultures of UG30, greater than 90% of 103 M 2,4-DNP was
transformed to other compds., while 8-19% of the 2,4-DNP was
mineralized within 5 days. A significant portion (20-50%) of the
2,4-DNP was metabolized to highly polar metabolite(s) with one major
unidentified metabolite accumulating from 5 to 25% of the initial
radioactivity. The amts. of 2,4-DNP mineralized and converted to
polar metabolites was affected by glutamate concn. in the medium.
Nitrophenolic compds. metabolized by UG30 were also suitable
substrates for the UG30 PCP-4-monooxygenase (pcpB gene expressed in Escherichia coli) which is likely central to degrdn. of these
compds. The wide substrate range of UG30 could render this strain
useful in bioremediation of some chem. contaminated soils.
132: 33042n Inhibition of trichothecin and ergosterol biosynthesis in Trichothecium roseum by lovastatin. Huang, WanLing; Lee, Kuan Rong; Shiao, Ming-Shi (Department of Life
Science, National Tsing Hua University, Hsinchu, Taiwan 300). J.
Chin. Chem. Soc. (Taipei) 1999, 46(5), 687-692 (Eng), Chinese
Chemical Society. The effect of lovastatin, an 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase inhibitor, on the biosynthesis of trichothecin, ergosterol, and fatty acids in the fungus Trichothecium
roseum was investigated. Treatment of lovastatin (50 M) to a
5-day-old culture of T. roseum reduced the incorporation of [2-14C]acetate into trichothecin by 79%, whereas the conversion of [5-3H]mevalonate into this sesquiterpenoid mycotoxin was reduced by only
28%. In a parallel expt., the incorporation of [2-14C]acetate and
[5-3H]mevalonate into ergosterol were decreased by 78% and 17%,
resp. Meanwhile, the conversion of labeled acetate into fatty acids
was not significantly affected. These results indicate that HMGCoA reductase is a major, but not strict, regulatory site in mevalonic
acid pathway leading to the formation of trichothecin and ergosterol.
No priority was fond in utilization of a single, residual mevalonic acid
pool in response to lovastatin inhibition for the biosynthesis of trichothecin and ergosterol. Inhibition of mevalonic acid formation does
not significantly divert acetyl-CoA into fatty acid synthesis.
132: 33044q Dual lipid modification of the yeast G subunit
Ste18p determines membrane localization of G. Hirschman,
Jodi E.; Jenness, Duane D. (Department of Molecular Genetics and
Microbiology, University of Massachusetts Medical School, Worcester,
MA 01655-0122 USA). Mol. Cell. Biol. 1999, 19(11), 7705-7711
(Eng), American Society for Microbiology. The pheromone response
in the yeast Saccharomyces cerevisiae is mediated by a heterotrimeric G protein. The G subunit (a complex of Ste4p and Ste18p) is
assocd. with both internal and plasma membranes, and a portion is
not stably assocd. with either membrane fraction. Like Ras, Ste18p
contains a farnesyl-directing CaaX box motif (C-terminal residues
107 to 110) and a cysteine residue (Cys 106) that is a potential site
for palmitoylation. Mutant Ste18p contg. serine at position 106 (mutation ste18-C106S) migrated more rapidly than wild-type Ste18p during sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDSPAGE). The electrophoretic mobility of wild-type Ste18p (but not
the mutant Ste18p) was sensitive to hydroxylamine treatment, consistent with palmitoyl modification at Cys 106. Furthermore, immunopptn. of the G complex from cells cultured in the presence of
[3H]palmitic acid resulted in two radioactive species on nonreducing
SDS-PAGE gels, with mol. wts. corresponding to G and G.
Substitution of serine for either Cys 107 or Cys 106 resulted in the
failure of G to assoc. with membranes. The Cys 107 substitution
also resulted in reduced steady-state accumulation of Ste18p, suggesting that the stability of Ste18p requires modification at Cys 107.
All of the mutant forms of Ste18p formed complexes with Ste4p, as
Issue 3, 2000
assessed by coimmunopptn. We conclude that tight membrane attachment of the wild-type G depends on palmitoylation at Cys 106
and prenylation at Cys 107 of Ste18p.
132: 33046s Degradation of desferrioxamines by Azospirillum irakense: assignment of metabolites by HPLC/electrospray
mass spectrometry. Winkelmann, G.; Busch, B.; Hartmann, A.;
Kirchhof, G.; Sussmuth, R.; Jung, G. (Department of Microbiology
and Biotechnology, University of Tubingen, Tubingen, Germany). BioMetals 1999, 12(3), 255-264 (Eng), Kluwer Academic Publishers.
Based on a recent finding that an Azospirillum isolate ASP-1 possessing high 16S rDNA similarity to Azospirillum irakense was able
to degrade desferrioxamine type siderophores (Winkelmann et al. BioMetals 9, 78-83, 1996), various members of the genus Azospirillum
were analyzed for their ability to degrade desferrioxamines. While
the desferrioxamine-degrading activity was absent or scarcely detectable in strains of A. lipoferum, A. brasilense, A. amazonense, degrdn.
activity seemed to be confined to the species A. irakense (KBC-1,
KA3). Also the identity of strain ASP-1 as A. irakense could be
confirmed by species-specific oligonucleotide hybridization, InterLINE PCR fingerprinting and carbon source utilization pattern (BIOLOG) anal. Products of desferrioxamine B degrdn. were analyzed
by anal. HPLC and HPLC/electrospray mass spectrometry. Using
whole cells and purified enzyme it was shown that the trihydroxamate desferrioxamine B (561 amu) is split at the N-terminal amide
bond yielding a monohydroxamate (MH1, 219 amu) and a dihydroxamate (DH1, 361 amu) metabolite. A second monohydroxamate (MH2,
319 amu) resulted from DH1 after splitting the acetylhydroxamate
bond. Minor amts. of a further dihydroxamate (DH2, 419 amu)
originated from splitting the second amide bond in desferrioxamine
B. In addn. to desferrioxamine B, several other linear and cyclic
desferrioxamines and derivs. were degraded, whereas desferricoprogen and desferri-ferrichrome were not degraded, indicating high
substrate specificity of the desferrioxamine hydrolase in A. irakense
species. A simple microtiter plate assay was developed which can be
used to phenotypically discriminate and identify species of A. irakense from other Azospirillum species by their characteristic feature
of desferrioxamine degrdn.
132: 33047t Peculiarities of carbohydrate catabolism in plasmid containing Staphylococcus strains. Kozitskaya, S. N.; Gavrilyuck, V. G.; Golodock, L. P.; Vinnikov, A. I. (Dniepropetrovsk State
University, Ukraine). Ukr. Biokhim. Zh. 1999, 71(3), 26-29 (Russ),
Institut Biokhimii im. A. V. Palladina NAN Ukrainy. It was established, that the antibiotic resistant strains of Staphylococcus aureus
differed from susceptible strains in the intensity of the catabolic reactions of carbohydrate transformation and in the accumulation of
metab. central products, that is a result the R-plasmid presence in
them.
132: 33048u Production of polygalacturonase and increase of
longitudinal gas permeability in southern pine by brown-rot
and white-rot fungi. Green, Frederick, III; Clausen, Carol A.
(Forest Service, Forest Products Laboratory, USDA, Madison, WI
53705 USA). Holzforschung 1999, 53(6), 563-568 (Eng), Walter de
Gruyter GmbH & Co. KG. Hydrolysis of bordered and pinoid pits
may be a key event during colonization of wood by decay fungi.
Although pits are numerous, studies of pectin-hydrolyzing enzymes
in wood decay fungi are scarce, probably because of the relatively low
content (less than 4%) of pectin in wood and because of the primary
focus on understanding the degrdn. of lignified components. Endopolygalacturonase (endo-PG) activity was estd. by cup-plate assay and
viscosity redn. of pectin from liq. cultures of fifteen brown-rot and
eight white-rot basidiomycetous fungi using sodium polypectate as
the carbon source. Oxalic acid was estd. in liq. culture and related
to mycelial wt. of each fungus. Changes in longitudinal gas permeability of southern pine cores exposed to selected decay fungi in liq.
culture were measured to det. the extent of hydrolysis of bordered
pits. Twelve of fifteen brown-rot and six of eight white-rot fungi
tested were pos. for at least one of the polygalacturonase test methods.
Accumulation of oxalic acid was detected in thirteen of fifteen brownrot isolates and none of the white-rot fungi tested. Gas permeability
of pine cores increased approx. fourfold among brown-rot fungi tested
and 18fold among white-rot fungi tested. SEM revealed bordered pit
membrane hydrolysis in cores colonized by white-rot fungi, but only
torus damage, weakening and tearing of the pit membranes, was

obsd. in cores exposed to brown-rot fungi. We conclude that both
brown- and white-rot decay fungi have the enzymic capacity to
hydrolyze pectin, damage bordered pit membranes, and increase wood
permeability during colonization and incipient decay.
132: 33137x Cloning and expression of the coat protein gene
of the FLA83 W strain of papaya ringspot virus. Mcmaster,
Russell J.; Boeshore, Maury L.; Tricoli, David M.; Reynolds, John
F.; Carney, Kim J.; Gonsalves, Dennis (Seminis Vegetable Seeds,
Issue 3, 2000
Page 13
Inc.; Cornell Research Foundation, Inc., USA) U.S. US 6,002,072

(Cl. 800-301; A01H5/00), 14 Dec 1999, WO Appl. 1995/US7,272, 7
Jun 1995; 48 pp., Cont.-in-part of U.S. Ser. No. 366,881, abandoned.
(Eng). The present invention relates to a coat protein gene of papaya
ringspot virus strain FLA83 W. This coat protein gene can be used
to prep. plants which are resistant to papaya ringspot virus. The
genes were cloned by RT-PCR and sense and antisense clones were
prepd. for Agrobacterium-mediated introduction into plants.
FERMENTATION & BIOINDUSTRIAL CHEMISTRY

132: 22187n Effect of volume ratio of internal aqueous phase
to organic membrane phase (w/o ratio) of water-in-oil emulsion on penicillin G extraction by emulsion liquid membrane.
Lee, S. C. (Kunsan, Miryong Dong, Department of Chemical Engineering, Kunsan National University, Chonbuk, S. Korea). J. Membr.
Sci. 1999, 163(2), 193-201 (Eng), Elsevier Science B.V.. The batch
extn. of penicillin G from a model media was studied so as to obtain
the optimal w/o ratio in an emulsion liq. membrane (ELM) system
with the help of our previous works. First of all, the effects of org.
solvents or surfactants in membrane phase on apparent degree of its
extn. at various w/o ratios were investigated. The org. solvents and
surfactants were kerosene/n-Bu acetate and Span 80/PARABAR 9551,
resp. The optimal compn. between two surfactants for the highest
apparent degree of extn. was obtained at each w/o ratio. With the
optimal surfactant compn. at each w/o ratio, citrate buffer solns. of
three different concns. having about pH 4.8 were selected as the
external aq. phase. The mass of Na2CO3 in the internal phase was
kept const. for every w/o ratio in order to investigate the effects of
w/o ratio on the degree of extn. under the same trapping mass of Na2CO3 for penicillin G. The highest possible concn. of Na2CO3 in the
internal phase was chosen using balance equations and equil. expressions. In consequence, the highest actual degree of extn. of 98.2%
and the lowest percentage of swelling of 36.0 were obtained in the
specific ELM system with 0.41 M citrate buffer soln., 0.175 M Na2CO3 and w/o ratio of 1/1.
132: 22188p Engineering a methymycin/pikromycin-calicheamicin hybrid: Construction of two new macrolides carrying a designed sugar moiety. Zhao, Lishan; Ahlert, Joachim; Xue,
Yongquan; Thorson, Jon S.; Sherman, David H.; Liu, Hung-wen
(Departments of Chemistry and Microbiology and Biological Process
Technology Institute, University of Minnesota, Minneapolis, MN 55455
USA). J. Am. Chem. Soc. 1999, 121(42), 9881-9882 (Eng), American
Chemical Society. Non-natural secondary metabolite glycosylation
O
Me
Me
Me
pds. (I and II) in yields of 11.0 and 1.5 mg, resp. Purified I and II
are inactive against S. pyogenes.
132: 22190h Amylose-like polysaccharide accumulation and
hyphal cell-surface structure in relation to citric acid production by Aspergillus niger in shake culture. Kirimura, K.; Yusa,
S.; Rugsaseel, S.; Nakagawa, H.; Osumi, M.; Usami, S. (Department of Applied Chemistry, School of Science and Engineering,
Waseda University, Tokyo, Japan 169-8555). Appl. Microbiol. Biotechnol. 1999, 52(3), 421-428 (Eng), Springer-Verlag. When 120
mg glucose/mL was used as a carbon source, in shake culture Aspergillus niger Yang no. 2 maximally produced only 15.4 mg citric acid/mL
but accumulated 3.0 mg extracellular polysaccharide/mL. The polysaccharide secreted by mycelia of Yang no. 2 in shake culture was
confirmed to be an amylose-like R-1,4-glucan by hydrolysis anal.
with acid, amylase and glucoamylase. However, in static cultures,
such as semisolid and surface cultures free from phys. stresses caused
by shaking damage, Yang no. 2 produced more citric acid but did not
accumulate the polysaccharide. With cultivation time in shake
culture, the amt. of extracellular polysaccharide and the viscosity of
the culture broth increased. The increase of shaking speed caused a
remarkable increase in the accumulation of extracellular polysaccharide, e.g. 11.2 mg extracellular polysaccharide/mL was accumulated
in the medium at a shaking speed of 200 rpm. The addn. of 2.0 mg
CM-cellulose (CMC)/mL as a viscous additive to the medium reduced
drastically the amt. of extracellular polysaccharide accumulated to
1.5 mg/mL, but increased the citric acid produced to 52.0 mg/mL.
However, intracellular polysaccharide accumulation kept up a steady
rate of 0.26 g/mg dried mycelium through the entire period of cultivation. The addn. of 3.0 mg polysaccharide/mL purified from the culture
broth to the medium at the start of a culture resulted in a decrease
of extracellular polysaccharide accumulation but an increase of citric
acid accumulation. From electron-microscopic observation, cell
surfaces of hyphae cultivated with CMC were smooth, while hyphae
cultivated without CMC had fibrous and granular polysaccharide on
the cell surface. These results suggested that Yang no. 2 secreted the
polysaccharide on the cell surface as a viscous substance and/or a
shock absorber to protect itself from phys. stresses caused by shaking
damage in shake culture.
O
O
O
HO
Me
Me
Me
NHCOCH3
OH
O
Me
Me
Me
Me
O
O
HO
Me
O
O
Me
Me
NHCOCH3
OH
II
patterns are shown to be engineered through a rational selection of

heterologous gene combinations. The Streptomyces venezuelae
methymycin/pikromycin gene cluster was selected as the parent strain
and a gene (calH) from the calicheamicin biosynthetic gene cluster of
Micromonospora echinospora as the foreign collaborator gene. The
calH gene was amplified by PCR, cloned, and introduced by conjugal
transfer using Escherichia coli S 17-1 into a previously constructed
S. venezuelae mutant (KdesI) in which the desI gene was replaced
by the neomycin resistance gene. Transformants yielded the major
product 10-deoxymethynolide (400 mg) and 2 minor macrolide com-
132: 22193m Factors influencing the synthesis of polyethylene glycol400 oleate mono- and diesters by lipase. Kou, Xiufen;
Xu, Jiali (Institute of Microbiology, The Chinese Academy of Sciences, Beijing, Peop. Rep. China 100080). Shengwu Gongcheng Xuebao 1999, 15(3), 301-304 (Ch), Kexue Chubanshe. The factors
influencing the synthesis of polyethylene glycol400 oleate mono- and
diesters by immobilized lipase from Candida sp.-1619 were investigated. Mono- and diesters were formed after 6-h reaction with
different molar ratio of the substrates. The ratio of monoester to
diester formed was in the range of 3.5:1 to 4:1, when the molar ratio
of acid to PEG400 was from 0.25:1 to 2:1. Almost equal amts. of
mono- and diesters were produced when the molar ratio of acid to
PEG400 was 3:1 to 8:1. Only diester was found in reaction mixt. with
different molar ratio of the substrates when the equil. of reaction
have been reached (22 h). The ratio of mono- to diester was 1:3.2 in
the reaction system contg. hexane and the molar ratio of substrates
was 2:1.
132: 22194n Copolymerization of lignin with cresol catalyzed
by peroxidase in reversed micellar systems. Liu, Junhong;
Weiping, Yuan; Lo, Tao (Department of Chemical Engineering, Quingdao Institute of Chemical Technology, Tsingtao, Peop. Rep. China
266042). EJB Electron. J. Biotechnol. [online computer file] 1999,
Page 14
2(2), No pp. Given (Eng), Universidad Catolica de Valparaiso. Avail.

URL: http://www.ejb.org/content/vol2/issue2/full/4/reprint.asp Peroxidase-catalyzed copolymn. of lignin with cresol in a reversed micellar
system was performed successfully. The mol. wt. of the copolymer
was controlled by adjusting the surfactant concn., and a maximal
mean mol. wt. of 1890 kDa was obtained.
132: 22195p Production of (R)-3-pentyn-2-ol through stereoinversion of racemic 3-pentyn-2-ol by Nocardia fusca
AKU 2123. Xie, S.-X.; Ogawa, J.; Shimizu, S. (Division of Applied
Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto,
Japan 606-8502). Appl. Microbiol. Biotechnol. 1999, 52(3), 327-331
(Eng), Springer-Verlag. Wet cells of Nocardia fusca AKU 2123 are
good catalysts for the prodn. of (R)-3-pentyn-2-ol (PYOH) from
(RS)-PYOH through a stereoinversion reaction. Under optimal conditions (350 mM potassium phosphate buffer, pH 8.0, 30% (w/v) wet
cells, 0.12% NADPH, 10% glucose, and 30 U/mL glucose dehydrogenase) (R)-PYOH of high optical purity (98.7% e.e.) was produced
from 2% (vol./vol.) (RS)-PYOH with a yield of 70.4% by 140 h incubation.
132: 22196q Effect of pressurized solvents on ethanol production by the thermophilic bacterium Clostridium thermocellum. Knutson, B. L.; Strobel, H. J.; Nokes, S. E.; Dawson, K. A.;
Berberich, J. A.; Jones, C. R. (Department of Chemical and Materials Engineering, University of Kentucky, Lexington, KY 40546-0046
USA). J. Supercrit. Fluids 1999, 16(2), 149-156 (Eng), Elsevier Science B.V.. Inhibition of microbial metab. due to the presence of
metabolic products leads to reduced yields in some fermn. processes.
In-situ extn. of inhibitory fermn. products is one method to increase
the product yield. Solvent compatibility with the microorganism is
an important criterion in the selection of extractant solvents. Although supercrit. CO2 has been investigated for the post-fermn. extn.
of metabolic products, it has generally been rejected for in-situ extn.
due to its inhibition of the metab. of numerous microorganisms. The
objective of this study was to assess the impact of dense gases and
supercrit. fluids (nitrogen, CO2, and ethane) on the carbohydrate
consumption and ethanol formation by a model organism, Clostridium
thermocellum, a fibrolytic thermophilic bacterium. Non-growing cells
capable of metab. were incubated at 60C with cellobiose as a substrate
in the presence of the three pressurized fluids. The fermn. broth was
sampled with time for residual cellobiose and ethanol concn. The
rate and extent of ethanol prodn. were similar in cell suspensions
maintained at atm. pressure under nitrogen (conventional method)
and at 6.9 MPa under nitrogen. Ethane at 6.9 MPa reduced the
extent of ethanol prodn. by less than 20% relative to the atm. control,
whereas CO2 at the same pressure reduced ethanol formation by
more than 80%. These results suggest that pressurized hydrocarbons
may have benefits over supercrit. CO2 for the in-situ recovery of
volatile fermn. products.
Issue 3, 2000
Ohfuka, Yasuko (School of Human Environmental Sciences, Mukogawa, Women's University, Nishinmiya, Ikebiraki-cho, Japan 6638558). Mukogawa Joshi Daigaku Kiyo, Shizen Kagaku-hen 1998,
46, 51-55 (Eng), Mukogawa Joshi Daigaku. The fruits of Gardenia
jasminoides are used as the coloring agent for various foods. In this
paper, we report successful results in the induction of callus and
plant regeneration from flesh and leaf cultures. A high frequency of
embryogenic callus formation from fruit was obtained with the medium
contg. 0.01M of IAA and 0.01M of kinetin. On the other hand,
better results were obtained with a medium contg. more than 0.1M
of 2,4-D.
132: 22199t Biocompatibility of NaCS and PDADMAC microcapsules with Bacillus thuringiensis. Mei, Lehe; Yao, Shanjing;
Lin, Dongqiang; Cen, Peilin; Zhu, Ziqiang (Department of Biochemical Engineering, Zhejiang University, Hangzhou, Peop. Rep. China
310027). Huagong Xuebao (Chin. Ed.) 1999, 50(5), 592-597 (Ch),
Huaxue Gongye Chubanshe. A new polyelectrolyte complex microcapsule formed by interaction between NaCS and PDADMAC was
introduced. The effects of the concn. of NaCS and PDADMAC on the
cell growth and the consumption of glucose were studied. The immobilization of Bacillus thuringiensis using microcapsules of NaCS
and PDADMAC was studied primarily. The results showed that microcapsules of NaCS-PDADMAC have good biocompatibility with
Bacillus thuringiensis.
132: 22202p Preparation of cis-(1S,2R)-indanediol by the
microbial reduction of 1,2-indanedione. Chartrain, Michel M.;
Ikemoto, Norihiro; King, Anthony O. (Merck and Co., Inc., USA)
Brit. UK Pat. Appl. GB 2,336,841 (Cl. C07C39/14), 3 Nov 1999, US
Appl. 83,355, 28 Apr 1998; 32 pp. (Eng). Cis-(1S,2R)-indanediol (I)
OH
OH
H2 N
HO
II
useful for prepg. (1S)-amino-(2R)-indanol (II) is manufd. from 1,2indanediol with Trichosporon cutaneum. II is an useful intermediate
for manufg. HIV protease inhibitor Crixivan (or Indinavir). The
method does not give byproducts such as indanone and gives high
yield. The physiol. and morphol. characteristics of the T. cutaneum
were also given.
132: 22203q Antibiotic BE-65932 manufacture with Streptomyces. Nakase, Kazuo; Nakajima, Shigeru; Hirayama, Mioko;
Tanaka, Kenichi; Nagano, Rie; Kojiri, Katsuhisa; Suda, Hiroyuki
(Banyu Pharmaceutical Co., Ltd., Japan) Jpn. Kokai Tokkyo Koho
JP 11 349,590 [99 349,590] (Cl. C07D519/00), 21 Dec 1999, Appl.
1998/169,217, 2 Jun 1998; 8 pp. (Japan). The antibiotic BE-65932
NH2
132: 22197r Effect of harvesting protocol on performance of

a hollow fiber bioreactor. Gramer, Michael J.; Poeschl, Douglas
M.; Conroy, Mark J.; Hammer, Bruce E. (Cellex Biosciences, Inc.,
Minneapolis, MN USA). Biotechnol. Bioeng. 1999, 65(3), 334-340
(Eng), John Wiley & Sons, Inc.. In this study, a bioreactor subject
to Starling flow in closed shell batch harvest model was compared to
two forms of addnl. forced extracapillary (EC) space convection including EC circulation and EC cycling. Despite the presence of Starling
flow as the dominant EC convection mechanism in the batch harvest
system, the bioreactor start up was fairly good. However, the antibody
productivity of the batch harvest system fell off rapidly after day 20
resulting in only 4.5 g of antibody produced. EC circulation with flow
parallel to the fibers had a slightly better start up than the batch
harvest. However, the antibody productivity also dropped after day
20 with EC circulation, resulting in only 7.5 g of antibody produced.
EC cycling with flow both parallel and perpendicular to the fibers
resulted in a start up similar to that of EC circulation. However, in
contrast to the other two systems, antibody productivity in the EC
cycling system was stable over the 60-day expt. resulting in the
prodn. of 23 g of antibody. These results demonstrate the importance
of inducing the proper flow distribution in the EC space to allow
consistent and stable prodn. in hollow fiber bioreactors.
132: 22198s Plant regeneration from hypocotyl-derived calli
of Gardenia jasminoides. Okamura, Tokumitsu; Akino, Ritsuko;
MeOOC
HO
OMe
OH
O
O
H
C
HOH2 C
C
Et
(I) having a mol. formula of C24H31O11N5 is manufd. with Streptomyces sp. A 65932. Shake culture of the fungus, and isolation of I from
the mycelium by solvent extn. and chromatog. were shown. The
physiol. and morphol. characteristics of Streptomyces sp. A 65932,
and physicochem. characteristics of I were also given.
132: 22204r Biocatalytic process for the preparation of 3-0acyl-flavonoids. Nicolosi, Giovanni; Piatteli, Mario; Lambusta,
Daniela; Patti, Angela (Consiglio Nazionale Delle Ricerche; RaoErbe Di Rao Felice, Italy) PCT Int. Appl. WO 99 66,062 (Cl.
C12P17/06), 23 Dec 1999, Appl. 1998/EP3,736, 18 Jun 1998; 22 pp.
(Eng). The present invention provides a process for the prepn. of 3
monoesters of flavonoids, antioxidants, comprising the following steps:
(a) chem. esterification of flavonoid with an aliph. acyl group having
from 1 to 18 carbon atoms to give the corresponding peracetylated

flavonoid, or, alternatively, partially acylated flavonoid; (b) subsequent
alcoholysis with an aliph. alc. having from 1 to 8 carbon atoms, in
the presence of lipase from Mucor miehei in an org. solvent. Prepn.
of 3-O-palmitoylquercetin with Lipozyme IM was shown.
132: 22205s Polyol fatty ester manufacture and oil-in-water
emulsion. Gruening, Burghard; Hills, Geoffrey (Goldschmidt, Th.,
A.-G., Germany) Jpn. Kokai Tokkyo Koho JP 11 346,791
[99 346,791] (Cl. C12P7/62), 21 Dec 1999, DE Appl. 19,821,851, 15
May 1998; 7 pp. (Japan). The polyol fatty acid ester emulsifier that
contains at least one primary alc. group and one secondary alc. group
is manufd. by formation of polyol fatty acid ester, incubation with
enzyme that specifically cleave the primary alc. group. The method
promotes the content of the secondary alc. in the polyol fatty acid
ester and improves the HLB value of the polyol fatty acid ester.
132: 34784t Sodium glutamate. Sugimoto, Masahiro (Kyowa
Hakko K. K., Japan). Kagaku Purosesu 1998, 234-242 (Japan) Tokyo
Kagaku Dojin: Tokyo, Japan. A review with 13 refs. on development
of prodn. technique of sodium glutamate by fermn.
132: 34787w Non-conventional hydrolase chemistry: amide
and carbamate bond formation catalyzed by lipases. Gotor, V.
(Facultad de Quimica, Departamento de Quimica Organica e Inorganica, Universidad de Oviedo, Oviedo, Spain). Bioorg. Med. Chem.
1999, 7(10), 2189-2197 (Eng), Elsevier Science Ltd.. A review with
104 refs. Biocatalysis in nonaq. media is becoming increasingly
important in org. synthesis. Lipases are the most used enzymes, esp.
in transesterification reactions. However, in the last years the amidation reaction catalyzed by lipases has also been shown to be a
useful tool for the org. chemists. In this review, we discuss the possibilities of the enzymic aminolysis and ammonolysis reactions for the
prepn. of different amides and for the resoln. of esters, amines and
aminoalcs. The enzymic alkoxycarbonylation of amines opens a new
way for the synthesis of chiral carbamates.
132: 34807c Successful design and development of genetically engineered Saccharomyces yeasts for effective cofermentation of glucose and xylose from cellulosic biomass to fuel
ethanol. Ho, Nancy W. Y.; Chen, Zhengdao; Brainard, Adam P.;
Sedlak, Miroslav (Molecular Genetics Group, Laboratory of Renewable Resources Engineering, Purdue University, West Lafayette, IN
47907-1295 USA). Adv. Biochem. Eng./Biotechnol. 1999, 65(Recent
Progress in Bioconversion of Lignocellulosics), 163-192 (Eng),
Springer-Verlag. A review with 35 refs. Ethanol is an effective,
environmentally friendly, nonfossil, transportation biofuel that produces far less pollution than gasoline. Furthermore, ethanol can be
produced from plentiful, domestically available, renewable, cellulosic
biomass. However, cellulosic biomass contains two major sugars,
glucose and xylose, and a major obstacle in this process is that Saccharomyces yeasts, traditionally used and still the only microorganisms currently used for large scale industrial prodn. of ethanol from
glucose, are unable to ferment xylose to ethanol. This makes the use
of these safest, most effective Saccharomyces yeasts for conversion of
biomass to ethanol economically unfeasible. Since 1980, scientists
worldwide have actively been trying to develop genetically engineered
Saccharomyces yeasts to ferment xylose. In 1993, we achieved a
historic breakthrough to succeed in the development of the first genetically engineered Saccharomyces yeasts that can effectively ferment
both glucose and xylose to ethanol. This was accomplished by carefully redesigning the yeast metabolic pathway for fermenting xylose
to ethanol, including cloning three xylose-metabolizing genes, modifying the genetic systems controlling gene expression, changing the
dynamics of the carbon flow, etc. As a result, our recombinant yeasts
not only can effectively ferment both glucose and xylose to ethanol
when these sugars are present sep. in the medium, but also can
effectively coferment both glucose and xylose present in the same
medium simultaneously to ethanol. This has made it possible because
we have genetically engineered the Saccharomyces yeasts as such
that they are able to overcome some of the natural barrier present in
all microorganisms, such as the synthesis of the xylose metabolizing
enzymes not to be affected by the presence of glucose and by the
absence of xylose in the medium. This first generation of genetically
engineered glucose-xylose-cofermenting Saccharomyces yeasts relies
on the presence of a high-copy-no. 2-based plasmid that contains
the three cloned genetically modified xylose-metabolizing genes to
Issue 3, 2000
Page 15
provide the xylose-metabolizing capability. In 1995, we achieved

another breakthrough by creating the super-stable genetically engineered glucose-xylose-confermenting Saccharomyces yeasts which
contain multiple copies of the same three xylose-metabolizing genes
stably integrated on the yeast chromosome. This is another crit.
development which has made it possible for the genetically engineered
yeasts to be effective for cofermenting glucose and xylose by continuous fermn. It is widely believed that the successful development of
the stable glucose-xylose-cofermenting Saccharomyces yeasts has
made the biomass-to-ethanol technol. a step much closer to commercialization. In this paper, we present an overview of our rationales
and strategies as well as our methods and approaches that led to the
ingenious design and successful development of our genetically
engineered Saccharomyces yeasts for effective cofermentation of
glucose and xylose to biofuel ethanol.
132: 34814c Downstream processing of oxidative decarboxylated carbohydrates - comparison of ion exchange and electrodialysis. Elseviers, Myriam; Coomans, Sonia; Lemmens, Hilde;
Roper, Harald; Lutin, Florence (Eridania Beghin-Say Vilvoorde
Research & Development Centre, B-1800 Vilvoorde, Belg.). Pharm.
Technol. Eur. 1999, 11(6), 28, 30, 34, 36-39 (Eng), Advanstar Communications Inc.. Downstream processing of oxidative decarboxylated carbohydrates is currently performed by ion exchange chromatog.
Pretreatment in batch, before loading onto a continuous column battery, is required because of the high concn. of carbonates. The pretreated syrups, however, still contain high concns. of salts, which
reduces the efficiency of the subsequently used ion exchange resins.
Electrodialysis was evaluated as an alternative downstream treatment, and proved to be a reliable method of purifying the syrups
without pretreatment.
132: 34815d Recovery of lactic acid from aqueous model solutions and fermentation broths. Von Frieling, P.; Schugerl, K.
(Haltermann AG, 21107 Hamburg, Germany). Process Biochem.
(Oxford) 1999, 34(6,7), 685-696 (Eng), Elsevier Science Ireland Ltd..
The aim of the investigations was to recover lactic acid from fermn.
broths in the presence of citric acid and acetic acid. For this purpose
the reactive extn. of lactic acid was investigated from model media
and fermn. broths with various carrier/modifier/solvent combinations.
Expts. were performed using a secondary amine (Hoe F 2562), a
tertiary amine (Hostarex A 327) as well as a phosphine oxide as
carriers and butylacetate and kerosene as solvents. The effects of the
type and concn. of the carrier and modifier and the temp. on the
equil. distribution were evaluated. Relationships for the equil.
distribution were developed and the calcd. data were compared with
the measured data. In the case of non-polar kerosene solvent, good
agreement was found between the calcd. and measured values. With
the polar solvent, butylacetate, the phys. extn. also had to be taken
into account. By means of the developed relationships the adduct
formation consts. were detd. for the extn. systems investigated. In
order to investigate the selectivity of the carriers, tests were also
performed to sep. lactic acid from acetic and citric acid in model
media and in fermn. broths. The sepn. of the acids is only possible
in a multistage process. In situ extn. of lactic acid was not possible
because of the toxicity of the carriers and modifiers.
132: 34817f Performance of a two-stage fermentor with cell
recycle for continuous production of lactic acid. Nishiwaki, A.;
Dunn, I. J. (Department of Materials Chemistry and Bioengineering,
Oyama National College of Technology, Oyama, Japan 323). Bioprocess Eng. 1999, 21(4), 299-305 (Eng), Springer-Verlag. The performance of a recycle two-stage fermentor with cell separators after
each stage is analyzed numerically for continuous prodn. of lactic
acid. In this system, the bleed broth withdrawn from the first stage
is provided to the second fermentor to reuse viable cells in the bleed.
Biol. rate expressions and parametric values are taken from the
literature. The effects of operating parameters on the concns. of total
and viable cells, substrate and product in each stage, the lactic acid
productivity and the substrate conversion are examd. and discussed.
With respect to overall productivity and conversion, it is found that
the present fermentor system is more efficient than a conventional
chemostat fermentor with cell recycle.
132: 34818g Genetic engineering of Escherichia coli for the
production of precorrin-3 in vivo and in vitro. Roessner, C. A.;
Page 16
Park, J.-H.; Scott, A. I. (Department of Chemistry, Texas A&M

University, College Station, TX USA). Bioorg. Med. Chem. 1999, 7(10),
2215-2219 (Eng), Elsevier Science Ltd.. The construction of a new
recombinant strain of Escherichia coli in which two vitamin B12 biosynthetic genes, cobA and cobI, from Pseudomonas denitrificans are
simultaneously overexpressed has resulted in the in vivo synthesis
and accumulation of Factor III, an isobacteriochlorin not normally
synthesized in E. coli. A lysate of the new strain can take the place
of two lysates normally required to provide uroporphyrinogen III methyltransferase (cobA) and precorrin-2 methyltransferase (cobI) in
an anaerobic five-enzyme synthesis of the early B12 intermediate,
precorrin-3 (the reduced form of Factor III) from -aminolevulinic
acid.
132: 34819h Overproduction of L-cysteine and L-cystine by
expression of genes for feedback inhibition-insensitive serine
acetyltransferase from Arabidopsis thaliana in Escherichia
coli. Takagi, H.; Awano, N.; Kobayashi, S.-i.; Noji, M.; Saito, K.;
Nakamori, S. (Department of Bioscience, Fukui Prefectural University, Fukui, Japan). FEMS Microbiol. Lett. 1999, 179(2), 453-459
(Eng), Elsevier Science B.V.. Two cDNAs encoding feedback inhibition-insensitive serine acetyltransferases of Arabidopsis thaliana were
expressed in the chromosomal serine acetyltransferase-deficient and
L-cysteine non-utilizing Escherichia coli strain JM39-8. The transformants produced 1600 to 1700 mg L-1 of L-cysteine and L-cystine
from glucose. The amt. of these amino acids produced per cell was
30 to 60% higher than that of an E. coli strain carrying mutant serine
acetyltransferase less sensitive to feedback inhibition.
132: 34820b Thermostable nitrilase catalyzed production of
nicotinic acid from 3-cyanopyridine. Almatawah, Q. A.; Cowan,
D. A. (Department of Biochemistry and Molecular Biology, University
College London, London, UK). Enzyme Microb. Technol. 1999, 25(89), 718-724 (Eng), Elsevier Science Ireland Ltd.. A thermostable
nitrilase produced by the thermophilic bacterium Bacillus pallidus
Dac521 catalyzed the direct hydrolysis of 3-cyanopyridine to nicotinic
acid without detectable formation of nicotinamide. The reaction conditions for nicotinic acid prodn. were optimized by using free bacterial
cells. Temp. and pH optima were 60C and 8.0, resp., with no detectable mass transfer limitation at the highest cell loading. Under
optimized conditions, 100% of the 3-cyanopyridine substrate could be
converted to nicotinic acid at a conversion rate of 76 nmol/min/mg dry
cell wt. Free bacterial cells were effective in converting 3-cyanopyridine at concns. of up to 0.3 M and the intracellular 3-cyanopyridinase stability was increased in the presence of the substrate at concns.
of 0.2 and 0.3 M. Both 3-cyanopyridine and nicotinic acid inhibited
the hydrolysis of 3-cyanopyridine at concns. greater than 0.2 M.
Cells immobilized in calcium alginate beads retained 98% of initial
activity and were more resistant to inactivation/inhibition than nonimmobilized cells at 60C. Calcium alginate immobilized cells used
in a column bioreactor retained 100% of 3-cyanopyridinase activity
for over 100 h and 10 h when continuously supplied with 0.1 M
3-cyanopyridine at 50C and 60C, resp. The conversion efficiencies
of the bioreactors operated at 50C and 60C, at 100% 3-cyanopyridinase activity, were 104 mg (substrate)/g (cells)/h and 208 mg
(substrate)/g (cells)/h, resp.
132: 34821c Bioconversion of progesterone by the activated
immobilized conidia of Aspergillus ochraceus TS. Dutta, Tapan
K.; Samanta, Timir B. (Marine Biology Institute, Kamaishi City,
Japan 026). Curr. Microbiol. 1999, 39(6), 309-312 (Eng), SpringerVerlag New York Inc.. Progesterone was transformed to its 11Rhydroxy deriv. (100% e.e) by the activated immobilized conidia of
Aspergillus ochraceus TS. The immobilized prepn. retained 79% of
free conidial activity. The immobilized conidia, activated by nutrients,
exhibited an increase in 11R-hydroxylation, and it was free of the
side product 6, 11R-dihydroxy progesterone. The half life and
turnover of immobilized and activated immobilized conidia were 14
and 12 days and 187 and 416 moles of the product/g of conidia resp.
The pH and temp. profiles of the free conidia remained unaltered
after immobilization and activation. Some germination of conidia
inside the matrix owing to incubation with nutrients was detected by
SEM.
132: 34822d Effect of shear on morphology and erythromycin
production in Saccharopolyspora erythraea fermentations.
Issue 3, 2000
Heydarian, S. M.; Mirjalili, N.; Ison, A. P. (Department of Biochemical Engineering, The Advanced Centre for Biochemical Engineering,
University College London, London, UK WC1E 7JE). Bioprocess Eng.
1999, 21(1), 31-39 (Eng), Springer-Verlag. A complex medium was
used to investigate the effects of shear on the S. erythraea fermn. at
a 7 L scale. Maximum biomass was 11.1 ( 0.5 g L-1 at 1250 rpm
(tip speed ) 4.45 ms-1), whereas it was 12.7 ( 0.2 g L-1 at 350 rpm
(tip speed ) 1.07 ms-1). Specific erythromycin prodn. was not stirrer
speed dependent in the range of 350 to 1000 rpm and decreased by
10% at stirrer speed of 1250 rpm. Morphol. measurements using
image anal. showed that the major axis of the mycelia (both freely
dispersed and clumps) decreased after the end of the rapid growth
phase to a relatively const. value (equil. size) dependent on the stirrer
speed. The mech. properties of the cell wall were examd. by disruption of fermn. broth in a homogenizer, and it was shown that mech.
strength of the cell wall increased to a large extent during deceleration phase.
132: 34823e Production of diosgenin and phytosterols in cell
suspension cultures of Dioscorea balcanica. Savikin-Fodulovic,
K.; Grubsic, D.; Culafic, Lj.; Menkovic, N. (Institute for Medicimal
Plants Research, 11000 Belgrade, Yugoslavia). Fiziol. Biokhim. Kul't.
Rast. 1999, 31(3), 173-178 (Eng), Izdatel'stvo "Logos". The possibility to initiate cell suspension was tested for five embryo-derived
callus lines of Dioscorea balcanica. Cell suspension cultures were
initiated only from soft and friable callus line E and consisted of
single spherical or elongated cells and small cell aggregates. Cell
suspension cultures were capable of diosgenin prodn. reaching the
max. content on the 5th week of cultivation (0.41 %). Among phytosterols, stigmasterol was the major compd. with the greatest amt.
on the 6th week of cultivation (0.16 %). The addn. of cholesterol, a
precursor in the biosynthesis of steroidal compds., increased the amt.
of both diosgenin and sitosterol while the effect of norflurazon which
is an inhibitor of carotenoid biosynthesis, was not significant.
132: 34824f Optimization of an Aspergillus niger glucose oxidase production process. Rothberg, A.; Weegar, J.; von Schalien,
R.; Fagervik, K.; Rydstrom, M.; Lind, K. (Faculty of Chemical
Engineering at Abo Akademi University, FIN-20500 Turku, Finland).
Bioprocess Eng. 1999, 21(4), 307-312 (Eng), Springer-Verlag. The
purpose of the present study was to ascertain the optimal concn. of
dissolved oxygen in order to maximize the intracellular glucose oxidase formation in Aspergillus niger. Cultivations performed in a
3.5-L lab. reactor showed that a dissolved oxygen concn. at 3% of
satn. at a total pressure of 1.2 bar was optimal for maximizing intracellular glucose oxidase activity. Cultivations performed at higher
dissolved oxygen concns. did not produce as much glucose oxidase as
those performed at 3%, although the formation rate was high. Expts.
revealed that maximal intracellular glucose oxidase formation for the
A. niger strain used, is accomplished by limiting the gluconic acid
prodn. rate by means of maintaining a low dissolved oxygen concn.
Several attempts to achieve higher intracellular glucose oxidase activity were also made by manipulating the glucose concn. at a 3% dissolved oxygen concn. However, no enhancement in glucose oxidase
activity was obsd.
132: 34825g Preparation and characterization of urease immobilized onto porous chitosan beads for urea hydrolysis.
Chen, J. P.; Chiu, S. H. (Department of Chemical Engineering, Chang
Gung University, Taoyuan, Taiwan 333). Bioprocess Eng. 1999, 21(4),
323-330 (Eng), Springer-Verlag. Urease was covalently immobilized
onto porous chitosan beads via primary amine groups connected to
the backbone via a six-carbon linear alkyl spacer. The optimum
conditions for enzyme immobilization are activating the beads with
1% (wt./wt.) glutaraldehyde, reacting the activated beads in pH 7
buffer with the enzyme, using an enzyme to bead wt. ratio of 25, and
without lyophilization. Chitosan-bound urease was found to fully
retain its specific activity. Properties of the immobilized urease were
characterized under batch and flow conditions. Increased optimum
reaction temp., enhanced thermal stability and storage stability, and
excellent reusability were found after enzyme immobilization. Continuous hydrolysis of urea soln. was studied in a column packed with
the enzyme-contg. beads for its possible application in regenerating
dialyzate soln. during hemodialysis.

132: 34832g Thermostable alkaline protease from Bacillus
brevis and its characterization as a laundry detergent additive. Banerjee, Uttam Chand; Sani, Rajesh Kumar; Azmi, Wamik;
Soni, Raman (Biochemical Engineering Research and Process Development Center, Institute of Microbial Technology, Chandigarh, 160036
India). Process Biochem. (Oxford) 1999, 35(1,2), 213-219 (Eng),
Elsevier Science Ireland Ltd.. An alk. protease from a facultatively
thermophilic and alkalophilic strain of B. brevis was studied. The
enzyme from a shake flask culture displayed max. activity at pH 10.5
and 37. The extracellular prodn. of the enzyme, its thermostable
nature and compatibility with most com. detergents are features which
suggest its application in detergent industry. The organism utilized
several C sources for the prodn. of proteases, lactose being the best
substrate followed by glucose and sucrose. Among the various org.
N sources, soybean meal was found to be the best. The protease was
stable at 25 for 288 h whereas, at 50 and 60, the half-lives were
60 and 7 h, resp. The thermostability of the protease was enhanced
by modifying its microenvironment. Acetate salts of Ca and Na
increased thermostability and protected against autolysis. The addn.
of Ca2+ (10 mM) and glycine (1M) individually and in combination
was found to be effective in increasing the half-life of the protease
many-fold. The enzyme retained >50% activity after 4 days at 60
in the presence of both Ca2+ (10 mM) and glycine (1M). The enzyme
showed compatibility at 60 with a no. of com. detergents in the
presence of Ca2+ and glycine. The enzyme improved the cleaning
power of various detergents. It could remove blood stains completely
when used with detergents in the presence of Ca2+ and glycine.
132: 34833h Engineering of a novel biochemical pathway for
the biosynthesis of l-2-aminobutyric acid in Escherichia coli
K12. Fotheringham, I. G.; Grinter, N.; Pantaleone, D. P.; Senkpeil,
R. F.; Taylor, P. P. (Biosciences, NSC Technologies, Mt. Prospect,
IL USA). Bioorg. Med. Chem. 1999, 7(10), 2209-2213 (Eng), Elsevier
Science Ltd.. l-2-Aminobutyric acid was synthesized in a transamination reaction from l-threonine and l-aspartic acid as substrates
in a whole cell biotransformation using recombinant Escherichia coli
K12. The cells contained the cloned genes tyrB, ilvA and alsS which
resp. encode tyrosine aminotransferase of E. coli, threonine deaminase of E. coli and R-acetolactate synthase of B. subtilis 168. The
2-aminobutyric acid was produced by the action of the aminotransferase on 2-ketobutyrate and l-aspartate. The 2-ketobutyrate is
generated in situ from l-threonine by the action of the deaminase,
and the pyruvate byproduct is eliminated by the acetolactate synthase. The concerted action of the three enzymes offers significant
yield and purity advantages over the process using the transaminase
alone with an eight to tenfold increase in the ratio of product to the
major impurity.
132: 34834j 5-Cyanovaleramide production using immobilized Pseudomonas chlororaphis B23. Hann, E. C.; Eisenberg,
A.; Fager, S. K.; Perkins, N. E.; Gallagher, F. G.; Cooper, S. M.;
Gavagan, J. E.; Stieglitz, B.; Hennessey, S. M.; DiCosimo, R.
(Experimental Station, DuPont Life Sciences, Wilmington, DE USA).
Bioorg. Med. Chem. 1999, 7(10), 2239-2245 (Eng), Elsevier Science
Ltd.. A biocatalytic process for the hydration of adiponitrile to 5-cyanovaleramide has been developed which can be run to higher conversion, produces more product per wt. of catalyst, and generates significantly less waste products than alternate chem. processes. The
biocatalyst consists of Pseudomonas chlororaphis B23 microbial cells
immobilized in calcium alginate beads. The cells contain a nitrile
hydratase (EC 4.2.1.84) which catalyzes the hydration of adiponitrile
to 5-cyanovaleramide with high regioselectivity, and with less than
5% selectivity to byproduct adipamide. Fifty-eight consecutive batch
reactions with biocatalyst recycle were run to convert a total of 12.7
metric tons of adiponitrile to 5-cyanovaleramide. At 97% adiponitrile conversion, the yield of 5-cyanovaleramide was 13.6 metric tons
(93% yield, 96% selectivity), and the total wt. of 5-cyanovaleramide
produced per wt. of catalyst was 3150 kg/kg (dry cell wt.).
132: 34835k Biotransformations in two-liquid-phase systems. Production of phenylacetaldehyde by oxidation of 2-phenylethanol with acetic acid bacteria. Molinari, F.; Gandolfi, R.;
Aragozzini, F.; Leon, R.; Prazeres, D. M. F. (Sezione Microbiologia
Industriale, Dipartimento di Scienze e Tecnologie Alimentari e Microbiologiche, Universita degli Studi di Milano, 20133 Milan, Italy).
Enzyme Microb. Technol. 1999, 25(8-9), 729-735 (Eng), Elsevier
Issue 3, 2000
Page 17
Science Ireland Ltd.. Phenylacetaldehyde can be obtained by oxidn.

of 2-phenylethanol with acetic acid bacteria in two-liq.-phase
systems where the aldehyde is removed into the org. phase before its
further conversion to acid. Two Acetobacter strains (ALEF and ALEG)
were able to accumulate aldehyde when aliph. hydrocarbons were
used. A two-liq.-phase system, composed of water and isooctane
(vol./vol., 1/1), was particularly suited for a significant accumulation
of the aldehyde: Acetobacter sp. ALEG furnished 9 g/l of phenylacetaldehyde within 4 h starting from 10 g/l of alc. and still 8 g/l were
recovered after 24 h in the org. phase, whereas strain ALEF gave 3.5
g/l of aldehyde from 5.0 g/l of substrate. Acetobacter sp. ALEG also
showed satisfactory long-term stability, being able to perform the
transformation with 80% of the original activity after 3 days of contact
with the solvent.
132: 34836m Part I. Enzymatic synthesis of lactate and glycolate esters of fatty alcohols. Torres, C.; Otero, C. (Departamento
de Biocatalisis, Instituto de Catalisis, CSIC, Cantoblanco, Madrid,
Spain 28049). Enzyme Microb. Technol. 1999, 25(8-9), 745-752
(Eng), Elsevier Science Ireland Ltd.. Optimum conditions were detd.
for the esterification reactions of lactic and glycolic acids with fatty
alcs. (C8-C16) in the presence of a lipase from Candida antarctica.
This synthetic method gives nearly complete conversion to the desired
ester in a relatively short time with high volumetric productivity. In
acetonitrile the max. yields of dodecyl lactate (95% in 48 h) and glycolate (87% in 24 h) were obtained in the presence of a desiccant and
0.28% (wt./wt.) added water, resp. The procedure permits one to
increase substrate concns. without significant adverse effects on the
yield of lactate ester for lactic acid concns. of 0.25 to 1 M. Similar
yields of lactate ester (94-96%) were obtained for alc. chain lengths
from C8 to C16. Although esterification of lactic acid with fatty alcs.
is not favored in apolar solvents (e.g. n-hexane), esterification of
glycolic acid in n-hexane produces high yields of the glycolate ester
(96% in 4 h). In the solvent-free system, esterification of lactic acid
with a fatty alc. requires the presence of desiccant from the beginning
of the process (yield of 70% in 48 h). For the reactions of glycolic
acid, a strategy in which a desiccant is added after 24 h of reaction
gives the max. yield of the glycolate ester (91%) in a shorter time (48
h of total reaction time). Alternatively, transesterification between
the alc. of interest and Et lactate increases the max. yield of dodecyl
lactate (87% in 24 h).
132: 34837n Closed-loop control of bacterial high-celldensity fed-batch cultures: production of mcl-PHAs by
Pseudomonas putida KT2442 under single-substrate and cofeeding conditions. Kellerhals, Michele B.; Kessler, Birgit; Witholt,
Bernard (Institute of Biotechnology, ETH Honggerberg, CH-8093
Zurich, Switz.). Biotechnol. Bioeng. 1999, 65(3), 306-315 (Eng), John
Wiley & Sons, Inc.. P. putida KT2442 is able to accumulate mediumchain-length poly(3-hydroxyalkanoates) (mcl-PHAs) as intracellular inclusions on a variety of fatty acids and many other carbon
sources. Some of these substrates, such as octanoic acid, alkenoic
acids, and halogenated derivs., are toxic when present in excess. Efficient prodn. of mcl-PHAs on such toxic substrates therefore requires
control of the C source concn. in the supernatant. In this study, a
closed-loop control system based on online gas chromatog. was
developed to maintain continuously fed substrates at desired levels.
The graphical programming environment LABVIEW was used to set
up a flexible process control system that allows users to perform
supervisory process control and permits remote access to the fermn.
system over the internet. Single-substrate supernatant concn. in a
high-cell-d. fed-batch fermn. process was controlled by a proportional (P) controller (P ) 50%) acting on the substrate pump feed
rate. Na-octanoate concns. oscillated around the setpoint of 10 mM
and could be maintained between 0 and 25 mM at substrate uptake
rates as high as 90 mmol/L-h. Under cofeeding conditions Na-10undecenoate and Na-octanoate could be individually controlled at
2.5 mM and 9 mM, resp., by applying a proportional integral controller for each substrate. The resulting copolymer contained 43.5 mol%
unsatd. monomers and reflected the ratio of 10-undecenoate in the
feed. It was suggested that both substrates were consumed at similar
rates. These results show that this control system is suitable for
avoiding substrate toxicity and supplying C substrates for growth
and mcl-PHA accumulation.
132: 34838p Production of erythritol from glucose by an osmophilic mutant of Candida magnoliae. Yang, Sung-Wook;
Page 18
Park, Jin-Byung; Han, Nam Soo; Ryu, Yeon-Woo; Seo, Jin-Ho

(Division of Chemical Engineering and Biotechnology, Ajou University,
Suwon, 442-749 S. Korea). Biotechnol. Lett. 1999, 21(10), 887-890
(Eng), Kluwer Academic Publishers. Candida magnoliae and its
mutants were analyzed to produce erythritol from glucose with high
yield and productivity. One mutant, M2, showed higher erythritol
conversion yield and productivity than the wild strain. The osmophilic mutant produced 25 g erythritol L-1 after 83 h of a flask culture
in a medium contg. 10% (w/v) glucose, corresponding to a 25% increase
in erythritol and a 30% increase in erythritol productivity compared
with the wild type. The fermn. properties were further improved by
cultivating the osmophilic mutant in a fermenter contg. 20% (w/v)
glucose medium with 0.54 g L-1 h-1 of erythritol productivity and
43% of erythritol conversion yield based on glucose.
132: 34839q Evaluation of xylitol production from corn cob
hemicellulose hydrolyzate by Candida parapsilosis. Kim, SangYong; Oh, Deok-Kun; Kim, Jung-Hoe (Dongcheon Consulting, Kyonggi-Do, 445-930 S. Korea). Biotechnol. Lett. 1999, 21(10), 891895 (Eng), Kluwer Academic Publishers. Candida parapsilosis was
grown for 59 h in a medium contg. corn cob hydrolyzate consisting of
50 g xylose L-1, 3.0 g glucose L-1, 2.0 g arabinose L-1, and 0.9 g
acetic acid L-1. A biomass of 9.1 g L-1 was produced with 36 g xylitol
L-1 and 2.5 g ethanol L-1. In a medium contg. 50 g xylose L-1
instead of corn cob hydrolyzate, the concns. of cells, xylitol, and ethanol were 8.6 g L-1, 33 g L-1, and 0.2 g L-1, resp. The differences
between two cultures were due to the glucose and arabinose in the
corn cob hydrolyzate stimulating growth and the low concn. of acetic
acid stimulating xylitol prodn.
132: 34840h Xylitol production from wood hydrolyzates by
entrapped Debaryomyces hansenii and Candida guilliermondii cells. Dominguez, Jose Manuel; Cruz, Jose Manuel; Roca, Enrique; Dominguez, Herminia; Parajo, Juan Carlos (Chemical Engineering Department, University of Vigo, Ourense, Spain 32004). Appl.
Biochem. Biotechnol. 1999, 81(2), 119-130 (Eng), Humana Press Inc..
Debaryomyces hansenii cells were entrapped in Ca-alginate beads
and used for producing xylitol from wood hydrolyzates. Batch expts.
showed that bioconversion was severely hindered when Ca-alginate
beads were hardened with Al3+ solns. As an alternative to Al3+
hardening, the improvements in both mech. stability of bioparticles
and fermenting ability of the immobilized system derived from using
increased concns. of sodium alginate were assessed. The best results
were obtained using a 4% (w/v) Na-alginate soln. in the gelification
step. This concn. was selected to perform continuous fermns. in a
packed-bed reactor using raw or charcoal-treated hydrolyzates (15.5
g of xylose/L) with two different yeasts: Candida guilliermondii and
Debaryomyces hansenii. With a final cell concn. of about 50 g of
cells/L (0.075 g of cells/g of beads), the volumetric productivities
reached with these yeasts in media made from charcoal-treated hydrolyzates were 0.58 and 0.91 g/Lh, resp.
132: 34841j Induction of alkaloid diversity in hybrid plant
cell cultures. Sheludko, Yu.; Gerasimenko, I.; Unger, M.; Kostenyuk, I.; Stoeckigt, J. (Institute Cell Biology Genetic Engineering,
Natl. Acad. Sci. Ukraine, Kiev, Ukraine 252143). Plant Cell Rep.
1999, 18(11), 911-918 (Eng), Springer-Verlag. The treatment of
Rauwolfia serpentina Rhazya stricta somatic hybrid cell suspension
culture with 100 M of Me jasmonate led to a general increase in
indole alkaloid content and to qual. changes in the alkaloid pattern.
The content of six alkaloids were investigated with respect to their
content in both the cell biomass and nutrition medium. Intracellular
17-O-acetyl-norajmaline content on the 5th day after treatment
had increased about 40-fold compared with the control culture. The
resp. concns. of the other alkaloids increased by a factor of two to
five. In total 26 indole alkaloids were identified in exts. of the Me
jasmonate-treated culture by TLC, UV, MS, and NMR data and
comparison with ref. alkaloids. The identification of macrophylline,
yohimbine oxindole, and yohimbine pseudoindoxyl has not been
reported before in Rauwolfia serpentina or Rhazya stricta plants nor
in cell cultures derived from these plants.
132: 34843m Tetracycline-regulated overexpression of glycosyltransferases in Chinese hamster ovary cells. Umana, Pablo;
Jean-Mairet, Joel; Bailey, James E. (Institute of Biotechnology,
ETH-Zurich, CH-8093 Zurich, Switz.). Biotechnol. Bioeng. 1999,
Issue 3, 2000
65(5), 542-549 (Eng), John Wiley & Sons, Inc.. The glycosylation
patterns of recombinant therapeutic glycoproteins can be engineered
by overexpression of glycosyltransferases in the host cells used for
glycoprotein prodn. Most prior glycosylation engineering expts. have
involved constitutive expression of cloned glycosyltransferases. Here
tetracycline-regulated expression of 2 glycosyltransferases, N-acetylglucosaminyltransferases III and V (GnTIII and GnTV), is used to
manipulate glycoform biosynthesis in Chinese hamster ovary (CHO)
cells and to study the effect of glycosyltransferase overexpression on
this host. The amt. of GnTIII and GnTV in these cells and the glycosylation patterns of several cellular glycoproteins could be controlled
simply by manipulating the concn. of tetracycline in the culture
medium. Using this system, it was found that overexpression of either
GnTIII or GnTV to high levels led to growth inhibition and was toxic
to the cells, indicating that this may be a general feature of glycosyltransferase overexpression. This phenomenon has not been reported
previously, probably due to the widespread use of constitutive promoters, and should be taken into account when designing vectors for
glycosylation engineering. The growth inhibition effect sets an upper
limit to the level of glycosyltransferase overexpression and may
thereby also limit the max. extent of in vivo modification of poorly
accessible glycosylation sites. Also, such inhibition implies a bound
on constitutive glycosyltransferase expression which can be cloned.
132: 34846q Antiapoptosis chemicals prolong productive lifetimes of mammalian cells upon Sindbis virus vector infection.
Mastrangelo, Alison J.; Zou, Shifa; Hardwick, J. Marie; Betenbaugh, Michael J. (Department of Chemical Engineering, The Johns
Hopkins University, Baltimore, MD 21218-2694 USA). Biotechnol.
Bioeng. 1999, 65(3), 298-305 (Eng), John Wiley & Sons, Inc.. Viral
expression systems allow for the rapid prodn. of large amts. of
recombinant protein in cell culture. In particular, Sindbis virus vectors now exist that make possible the expression of a variety of heterologous proteins in mammalian culture systems. Unfortunately,
infection of cultured cells with Sindbis virus vectors typically results
in apoptotic cell death, as demonstrated in the current study by DNA
laddering and fluorescence microscopy. Fortunately, it has recently
been demonstrated that apoptosis can be inhibited in vitro by certain
chem. reagents that are capable of blocking specific steps during the
cell death cascade. In this study, a rat prostate carcinomal cell line,
AT3-neo, was infected with a Sindbis virus vector contg. the gene for
chloramphenicol acetyltransferase (dsSV-CAT) in the presence of
several representative antiapoptotic chems. and analyzed for cell viability as well as recombinant protein prodn. N-acetylcysteine (NAC),
pyrrolidine dithiocarbamate (PDTC), bongkrekic acid (BA), and N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (Z-VAD.fmk) all
exhibited the capacity to limit apoptosis in the infected cells. Cells
maintained on these agents were able to survive the infection 1-3
days longer than the control samples. In addn. to providing gains in
cell viability, chem. treatment allowed for higher levels of recombinant
protein prodn. in most cases. Max. chloramphenicol acetyltransferase (CAT) productivities in cells maintained on BA, NAC, and
Z-VAD.fmk were 1.7-, 2.2-, and 3.9-fold higher than those obtained
from the untreated cultures. Consequently, the addn. of chem.
reagents to culture media as a means of inhibiting apoptosis may be
a valuable tool in the cell culture industry, where cell death severely
limits productivity levels and adds significantly to prodn. costs.
132: 34850m Rapid screening of solvents and carrier compounds for lactic acid recovery by emulsion liquid extraction
and toxicity on Lactobacillus casei (ATCC 11443). Demirci, Ali;
Pometto, Anthony L., III; Harkins, Kristi R. (Department of Food
Science and Human Nutrition, Iowa State University, Ames, IA 50011
USA). Bioseparation 1999, 7(6), 297-308 (Eng), Kluwer Academic
Publishers. This paper describes a rapid method to identify the best
solvent and carrier compd. combinations with the highest extn.
capability and the lowest microbial toxicity characteristics for product
recovery from microbial fermn. The extn. system has an aq. phase,
and an emulsion phase, which was a blend of sodium carbonate and
org. phase [91% (vol./vol.) org. solvent, 5% (vol./vol. or wt./v) carrier
compd., and 4% (vol./vol.) surfactant Span 80]. Alamine 336, or trin-octylamine in n-heptane; Alamine 336, Alamine 304, or tri-Bu
phosphate in hexane; and Alamine 304 or tri-Bu phosphate in isooctane; Alamine 304 or Amberlite in xylene demonstrated high lactic
acid extn. For detn. of bacterial toxicity of selected solvent and carrier compds., Lactobacillus casei subsp. rhamnosus (ATCC 11443)

was grown in LAF medium contg. one of the selected org. solvent,
carrier compd., and Span 80 in 250 mL flask at 37 and 125 rpm.
Samples were collected regularly during 48 h incubation, and measured for changes in cell d. by absorbance at 620 nm, cell count using
a fluorescent dye with flow cytometry, and lactic acid, and glucose
concns. by HPLC. Hexadecane:tributyl phosphate, n-dodecane:trin-octylamine, and kerosene:tri-n-octylphosphine oxide demonstrated the least microbial toxicity among the tested blends with excess
solvent media. Whereas, hexanes:Alamine 304 and xylenes:Alamine
304 were nontoxic in solvent satd. media.
132: 34851n Kinetic model for the bioconversion of glucose
to 2,5-diketo-D-gluconic acid. Zelic, B.; Pavlovic, N.; Delic, V.;
Vasic-Racki, D. (Faculty of Chemical Engineering and Technology,
University of Zagreb, Zagreb, Croatia HR-10000). Bioprocess Eng.
1999, 21(1), 45-50 (Eng), Springer-Verlag. A simple unstructured
math. model for the oxidn. of glucose to 2,5-diketo-D-gluconic acid
with Erwinia citreus was developed. The kinetic parameters of the
model were estd. by the oxygen partial pressure method and directly
from the temporal response obtained from the exptl. data collected in
the batch fermn. The simple developed model based on the kinetic
measurements was able to simulate quite well the dynamic behavior
of the batch fermn.
132: 34861r Refolding of a proinsulin hybrid polypeptide to
produce recombinant human insulin. Hartman, Jacob R.; Mendelovitz, Simona; Gorecki, Marian (Bio-Technology General Corp.,
USA) U.S. US 6,001,604 (Cl. 435-69.4; C07K1/107), 14 Dec 1999,
US Appl. 367,454, 29 Dec 1994; 28 pp., Cont. of U.S. Ser. No. 367,454,
abandoned. (Eng). An improved and efficient process for the prodn.
of recombinant human insulin by folding of an SOD-proinsulin hybrid
polypeptide is provided. Specifically claimed is a method of producing insulin which comprises: (a) obtaining a hybrid polypeptide
comprising proinsulin by treating a bacterial cell contg. DNA encoding the hybrid polypeptide, so that the hybrid polypeptide is expressed
and is recovered from the cell; (b) folding the hybrid polypeptide
without first subjecting the hybrid polypeptide to sulfitolysis under
conditions that permit correct disulfide bond formation in absence of
exogenous reducing agent; (c) subjecting the resulting folded, disulfide bonded hybrid polypeptide to enzymic cleavage to produce insulin;
and (d) purifying the insulin so produced.
132: 34862s FO-6903 A and B manufacture with Trichoderma. Omura, Satoru; Matsumoto, Tsukasa; Tomoda, Hiroshi;
Masuma, Rokuro; Shiomi, Kazuaki; Yamada, Haruki (Kitasato
Institute, Japan) Jpn. Kokai Tokkyo Koho JP 11 349,520
[99 349,520] (Cl. C07C49/707), 21 Dec 1999, Appl. 1998/163,114, 11
Jun 1998; 10 pp. (Japan). Antirheumatics FO-6903 A (I) and B (II)
OH
OH
CH Me
OH
CH Me
OH
OH
O
OH
II
are manufd. with Trichoderma. I and II are inhibitors to interleukin

1-converting enzyme and sphingomyelinase. Shake-culture of the
Trichoderma and isolation of these inhibitors from the culture supernatant were shown. The physiol. and morphol. characteristics of the
Trichoderma and physicochem. characteristics of I and II were also
given.
132: 34863t Antitumor substance BE-69785A and its manufacture. Shitakawa, Haruki; Nakajima, Shigeru; Hirayama, Mioko;
Kondo, Hisao; Ojiri, Katsuhisa; Suda, Hiroyuki (Banyu Pharmaceutical Co., Ltd., Japan) Jpn. Kokai Tokkyo Koho JP 11 349,522
[99 349,522] (Cl. C07C50/32), 21 Dec 1999, Appl. 1998/169,218, 2
Jun 1998; 7 pp. (Japan). The antitumor substance BE-69785A (I)
useful to control human and mouse tumors is manufd. with Streptomyces. Shake-culture of the Streptomyces and isolation of I from
Issue 3, 2000
Page 19
the mycelium by solvent extn. and chromatog. were shown. The physicochem. characteristics of I and physiol. and morphol. characteristics
of the Streptomyces were also given.
O
Me
Me
HO
Me
OH
Me
OH
132: 34864u Fermentation medium and method for producing r,-alkanedicarboxylic acids. Mobley, David Paul; Shank,
Gary Keith (General Electric Co., USA) U.S. US 6,004,784 (Cl.
435-134; C12P7/64), 21 Dec 1999, Appl. 152,386, 14 Sep 1998; 5 pp.
(Eng). This invention describes a low cost biofermentation medium,
and an economical method using the medium, for the manuf. of R,alkanedicarboxylic acids. The invention provides a biofermentation
medium and a method of bioprodn. for these important diacids which
makes their large scale com. prodn. economically feasible using a
biocatalyst. This method of prodn. obviates the need for chem.
synthesis using expensive starting materials from fossil fuels, and
does not generate a costly hazardous waste stream.
132: 34865v Steroid manufacture with recombinant CYP
gene-expressing Schizosaccharomyces. Bureik, Matthias; Bernhardt, Rita Germany Ger. Offen. DE 19,826,821 (Cl. C12N15/81),
23 Dec 1999, Appl. 19,826,821, 16 Jun 1998; 8 pp. (Ger). Recombinant
Schizosaccharomyces which express mitochondrial cytochrome P 450
and the use of these transformants to produce steroids are disclosed.
Thus, integration plasmid pt5-CYP11B2, contg. the aldosterone synthase gene CYP11B2, was prepd. and recombinant S. pombe transformed with this plasmid were created. This recombinant fission
yeast converted deoxycorticosterone to corticosterone, 18-hydroxycorticosterone, and aldosterone.
132: 34866w Treatment of maize brewing waste liquid. Wang,
Jianjun Peop. Rep. China Faming Zhuanli Shenqing Gongkai
Shuomingshu CN 1,175,627 (Cl. C12F3/00), 11 Mar 1998, Appl.
96,109,713, 5 Sep 1996; 6 pp. (Ch). The process comprises sepg. solid/
liq. by centrifugating, cooling liq. to <80, filtering with 20-30
hollow ultrafilter membrane to obtain solid and filtrate, feeding filtrate
after heat-exchanger back to mixt. process, and collecting the drying
solid for prepn. of protein fodder.
132: 34867x Extraction of alcohol from waste liquid containing sugar and acid. Zheng, Yixin; Pan, Mingkun; Xi, Yun; Lu,
Yonglin; Wang, Dehuai (Kunming Inst. of Environment Sciences,
Peop. Rep. China) Faming Zhuanli Shenqing Gongkai Shuomingshu CN 1,176,309 (Cl. C12P7/08), 18 Mar 1998, Appl. 97,109,865,
7 May 1997; 6 pp. (Ch). The process comprises mixing the waste liq.
contg. sugar and acid with starch, hydrolyzing at 120-130 and 0.10.15 MPa for 30-40 min to the sugar content of 10-15' BX, regulating pH to 4.2-4.5, removing ppt. by filtering, fermenting with yeast
such as beer yeast at 28-33 for 60-80 h twice, and prepg. alc. by
the traditional method. The waste liq. is obtained from the prodn. of
saponin from yellow ginger and yam. The starch is selected from
cassava with particle size of 0.5-1 mm, the addn. is 32-71 g/Kg.
132: 34868y Process for preparing erythritol using novel cell
of Pichia. Kim, Sang Yong; Oh, Deok Kun; Jung, Soo Ryun (Dong
Cheon Consulting Co., Ltd., S. Korea) U.S. US 6,001,616 (Cl. 435158; C12N1/16), 14 Dec 1999, Appl. 154,218, 16 Sep 1998; 6 pp. (Eng).
The present invention relates to a fermn. process to have a high
productivity with a novel mutant of Pichia sp., more specifically, for
prepg. erythritol under optimal fermn. conditions for max. erythritol
prodn. by optimizing the environmental conditions of culture such as
pH, temp. and by controlling osmotic pressure. A two-stage fermn.
was performed to control osmotic pressure. Osmotic pressure was
adjusted to a low level during growth phase and to a relatively high
level during prodn. phase by adding continuously glucose and NaCl
or KCl. Therefore, erythritol prodn. could be increased.
Page 20
Issue 3, 2000
MISCELLANEOUS
132: 22519x Accumulation of phycobiliproteins in cells of
ancient cyanobacteria from arctic permafrost as dependent
on the nitrogen source for growth. Erokhina, L. G.; Spirina, E.
V.; Gilichinskii, D. A. (Institute of Fundamental Problems of Biology, Russian Academy of Sciences, Pushchino, Russia 142292). Microbiology (Moscow) 1999, 68(5), 628-631 (Eng), MAIK Nauka/
Interperiodica Publishing. Anal. of the absorption spectra of cells
and phycobilisomes of ancient visible heterocystous cyanobacteria Nostoc sp. and Anabaena sp. isolated from frozen sediments of a Holocene lake and grown on various nitrogen sources revealed that the
amt. of C-phycoerythrin in relation to chlorophyll was the greatest
when cells were grown in nitrogen-free medium. In the presence of
nitrate salts or org. nitrogen sources such as asparagine and glutamine,
the content of C-phycoerythrin decreased. Conversely, the content
of phycocyanin increased in the presence of org. nitrogen sources.
Glycine inhibited the growth of cyanobacteria and reduced the content
of C-phycoerythrin and phycocyanin. Ammonium salts completely
inhibited growth. Accumulation of C-phycoerythrin was accompanied
by a decrease in the content of carotenoids. These results suggest
that the ancient cyanobacteria studied are similar to modern nitrogenfixing cyanobacteria in their capacity to fix mol. nitrogen and store it
in the form of C-phycoerythrin mols.
132: 22907x Synthesis and anti-microbial activity of pyrazolylindoles. Sonar, V. N.; Ali, Sadath; Hadimani, M. B. (Dept.
of Pharmaceutical Chemistry, V.L. College of Pharmacy, Raichur,
584101 India). Indian Drugs 1999, 36(8), 535-537 (Eng), Indian
Drug Manufacturers' Association. Substituted indole-2-carboxylates are refluxed with hydrazine hydrate (80%) to afford indole-2carboxyhydrazides. These hydrazides are treated with -diketones
such as acetylacetone, benzoylacetone, and dibenzoylmethane to get
2-(3',5-dimethylpyrazole-1'-carbonyl)indoles 2-(3'-methyl-5'phenylpyrazole-1'-carbonyl)indoles, and 2-(3',5'-diphenylpyrazole1-carbonyl)indoles resp. The compds. were subjected to antimicrobial
screening.
132: 23162n Selective hydrolysis of nucleotides to nucleosides and free bases. Chmielowiec, Urszula; Kruszewska, Hanna;
Cybulski, Jacek (Department of Chemistry, Pharmaceutical Research
Institute, 01-793 Warsaw, Pol.). Farmaco 1999, 54(9), 611-614
(Eng), Elsevier Science S.A.. The kinetics of the hydrolysis of 2'deoxyadenosine-5'-monophosphoric acid (dAMP), 2'-deoxycytidine5'-monophosphoric acid (dCMP), 2'-deoxyguanosine-5'-monophosphoric acid (dGMP) and thymidine-5'-monophosphoric acid (dTMP)
was studied in the presence of Xanthomonas maltophilia, Escherichia
coli, Pseudomonas putida and Lactobacillus acidophilus. The reaction products are nucleosides: 2'-deoxyadenosine (dA), 2'-deoxycytidine (dC), 2'-deoxyguanosine (dG) and thymidine (dT), resp., or
the resp. free bases. Hydrolysis of dTMP and dGMP proceeded stepwise according to the sequence: nucleotide f nucleoside f free base,
whereas no accumulation of the free base was obsd. during the hydrolysis of dAMP and dCMP.
132: 23192x Preparation of peptides having membraneinteracting property. Machida, Yukiko; Yu, Yung (Norinsuisansho Shokuhin Sogo Kenkyusho, Japan) Jpn. Kokai Tokkyo Koho
JP 11 335,394 [99 335,394] (Cl. C07K14/435), 7 Dec 1999, Appl.
1998/156,911, 22 May 1998; 11 pp. (Japan). The title peptides, in
particular H-Lys-Asn-Trp-Lys-Gly-Ile-Ala-Gly-Met-Ala-LysLys-Leu-Leu-Gly-Lys-Asn-Trp-Lys-Leu-Met-OH and H-LysAsn-Trp-Lys-Lys-Ile-Ala-Gly-Met-Ala-Lys-Lys-Leu-LeuLys-Lys-Asn-Trp-Lys-Leu-Met-OH, were prepd. by the solid
phase synthesis. These peptides exhibited specific capability for pore
formation in microbial membrane in animal cell membrane model but
no hemolytic activity against human red blood cells and are potentially
useful as antibacterial agents.
132: 24493h Application of petroleum microbial dewaxing
technology. Liu, Chenglin; Tan, Zhenming; Liu, Yuchao (Dushanzi General Petrochemical Works, Kelamayi, Peop. Rep. China
833600). Lianyou Sheji 1999, 29(9), 57-59 (Ch), Lianyou Sheji Bianjibu. A review with no ref. to introduce the technol. characteristics
of petroleum microbial fermn. dewaxing technol. and its application.
In comparison with the conventional solvent dewaxing process, the

microbial dewaxing has irreplaceable advantages of simple process
scheme, low investment cost of equipment, flexibility in feedstock
altering, and steady product quality. The pour point of the dewaxed
oil is <-40, and the yield is 90%.
132: 24736q Production of 2,3-butanediol by newly isolated
Enterobacter cloacae. Saha, B. C.; Bothast, R. J. (Fermentation
Biochemistry Research Unit, National Center for Agricultural Utilization Research, Agricultural Research Service, United States Department of Agriculture, Peoria, IL 61604 USA). Appl. Microbiol. Biotechnol. 1999, 52(3), 321-326 (Eng), Springer-Verlag. Enterobacter
cloacae NRRL B-23289 was isolated from local decaying wood/corn
soil samples while screening for microorganisms for conversion of
L-arabinose to fuel ethanol. The major product of fermn. by the
bacterium was meso-2,3-butanediol (2,3-BD). In a typical fermn.,
a BD yield of 0.4 g/g arabinose was obtained with a corresponding
productivity of 0.63 g/l per h at an initial arabinose concn. of 50 g/l.
The effects of initial arabinose concn., temp., pH, agitation, various
monosaccharides, and multiple sugar mixts. on 2,3-BD prodn. were
investigated. BD productivity, yield, and byproduct formation were
influenced significantly within these parameters. The bacterium
utilized sugars from acid plus enzyme saccharified corn fiber and
produced BD (0.35 g/g available sugars). It also produced BD from
dil. acid pretreated corn fiber by simultaneous saccharification and
fermn. (0.34 g/g theor. sugars).
132: 24771x Quality of solid biofuels - database and field
trials. Hartmann, H.; Maier, L.; Bohm, T. (Research Center of
Agricultural Engineering, Munich University of Technology, D-85354
Freising-Weihenstephan, Germany). Biomass, Proc. Biomass Conf.
Am., 4th 1999, 1, 273-279 (Eng). Edited by Overend, Ralph P.; Chornet, Esteban. Elsevier Science: Oxford, UK. Quality aspects of solid
biofuels were investigated in a new database. Most parameters varied
greatly, particularly when annually harvested biomass was considered.
For planning purposes the frequency distributions should be used
rather than mean values. The quality of some crops may be changed
by modified agricultural practices. Rainfall shortly after cutting can
deplete chlorine and potassium in grass by 60 to 80%.
132: 24782b Producing a life-cycle energy balance for an
integrated sweet sorghum / sugarcane system in the semiarid southeast region of Zimbabwe. Woods, J.; Hall, D. O. (Division of Life Sciences, King's College London, London, UK W8 7AH).
Biomass, Proc. Biomass Conf. Am., 4th 1999, 1, 387-392 (Eng).
Edited by Overend, Ralph P.; Chornet, Esteban. Elsevier Science:
Oxford, UK. During the last eight years we have carried out research
to establish the potential for energy prodn. from sweet stemmed varieties of the C4 crop sorghum [Sorghum bicolor (L.) Moench] that are
integrated with sugarcane. Sweet sorghum is a versatile crop capable
of producing large amts. of liq. fuels (i.e., ethanol by fermn.), and
electricity or thermal heat (by combustion of fibrous stem residues).
It is an annual energy crop that is tolerant to a wide range of environments and it efficiently utilizes water, nitrogen and light. A lifecycle energy balance methodol. has been designed as a part of the
development of a systems anal. model (AIP). The methodol. will be
used in the AIP to generate site-specific and time-specific ests. of
energy balances using both the integrated mechanistic sorghumspecific plant growth model (a modified CERES model) and the
harvesting, transport and energy conversion modules of the AIP. This
paper summarizes a worked example of an energy balance calcn.
based on a 1997/98 sweet sorghum productivity trial and the utilization of sweet sorghum and sugarcane for energy prodn. in the Triangle
Ltd. sugar mill in south-eastern Zimbabwe.
132: 35295w An in vivo 13C NMR analysis of the anaerobic
yeast metabolism of 1-13C-glucose. Giles, Brent J.; Matsche,
Zenziwe; Egeland, Ryan D.; Reed, Ryan A.; Morioka, Scott S.; Taber,
Richard L. (Department of Chemistry, The Colorado College, Colorado
Springs, CO 80903 USA). J. Chem. Educ. 1999, 76(11), 1564-1566
(Eng), Division of Chemical Education of the American Chemical
Society. An undergraduate biochem. lab. expt. using NMR to monitor, in real time, the anaerobic metab. of 1-13C-glucose by Saccha-

romyces cerevisiae (Fleishmann's Active Dry yeast), is presented. This
expt. is used to study how yeast maintains internal osmolarity when
subjected to osmotic shock by saline solns. It uses readily available
yeast, 1-13C-labeled glucose, and a stranded NMR spectrometer,
and students can complete the exercise in one lab. period.
132: 35332f Novel therapeutic agents for macromolecular
structures. Griffin, John H.; Moran, Edmund J.; Oare, David
(Advanced Medicine, Inc., USA) PCT Int. Appl. WO 99 64,036 (Cl.
A61K38/00), 16 Dec 1999, US Appl. 92,941, 15 Jul 1998; 209 pp.
(Eng). Title agents comprise a plurality of ligands (e.g., therapeutic
agents) linked by a linking structure such that said agents bind at
g1 site of a target structure (e.g., cellular, extracellular, and microbial
components derived from vectors, viruses, fungi, yeasts, bacteria, and
the like). Thus, (CH2)3(CONHCH2CH2NH2)2 was prepd. for amidation by a carboxy-contg. ligand.
132: 35333g Multibinding inhibitors of topoisomerase. Linsell, Martin S.; Meier-Davis, Susan; Griffin, John H. (Advanced
Medicine, Inc., USA) PCT Int. Appl. WO 99 64,054 (Cl. A61K38/00),
16 Dec 1999, US Appl. 93,072, 16 Jul 1998; 142 pp. (Eng). Novel
topoisomerase inhibitors that act as multibinding agents, LpXq [where
L ) a ligand capable of binding to topoisomerase; X ) a linker; p )
2-10; q ) 1-20; the distance between ligands 2-50 ], are disclosed.
Combinatorial arrays, methods of synthesis, and methods of assaying
the dimeric and multimeric compds. are also embodied by the invention. A no. of divalent prophetic examples, derived from substituted
fused ring heterocyclic ligands and difunctional linkers, are given.
Compds. of this invention are useful in the treatment and prevention
of cancer and microbial infections (no data). The multibinding compds. provide greater biol. and/or therapeutic effects than the aggregate of the unlinked ligands due to their multibinding properties
(no data). Ligands may include A-62176, A-74932, acridine carboxamides, actinomycin D, AD-312, AD-347, AHMA, AMP-53, amrubicin, amsacrine, anthracyclines, asulacrine, azonafide, azatoxin, BBR2778, BMY-43748, BO-2367, bromodeoxyuridine, C-1310, C-1311,
CC-131, CJ-12373, CI-937, CI-920 (fostriecin), CP-115953, camptothecin, daunorubicin, doxorubicin, DuP 937 (losoxathrone), DuP 941,
elinafide, ellipticine-estradiol (conjugates), elsamitrucin, ER-37328,
etoposide, fleroxacin, GI-149893, GL-331, GR-1222222X, ICRF154, ICRF-193, idarubicin, iododoxorubicin, IST-622, KRQ- 10018,
intoplicine, lomefloxacin, losoxantrone, m-AMSA, merbarone, meraboin, mitonafide, mitoxantrone, morindone, NCA-0465, NK-109,
NK-611, NSC-655649, NSC-665517, NSC-675967, pazelliptine, pazufloxacin, PD-131112, piroxantrone, pyridobenzophenoxazine,
S-16020-2, saintopin, sitafloxacin hydrate, SN-22995, sobuzoxane,
SR-103, TAS-103, teloxantrone, teniposide, TLC-D-99, top-53, topotecan, tosufloxacin, TRK-710, trovafloxacin, UCE-6, VM-26, VP16, W5R, WIN-33377, WIN-58161, WIN-645593, WQ-2743, WQ3034, WR-63320, XR-5942, XR-5000, and 773U82.
132: 35537b Synthesis and biological activities of phosphonothrixin. Nakamura, Kazuhiko; Kimura, Takashi; Takahashi, Eisaku (Department of Chemistry, Faculty of Science and Technology,
Keio University, Hiyoshi, Yokohama, Japan 223-0061). Phosphorus,
Sulfur Silicon Relat. Elem. 1999, 144-146, 613-616 (Eng), Gordon
& Breach Science Publishers. A novel herbicidal compd. phosphonoO
O
P(OH)2
OH
Me
OH
thrixin (I) possessing a C-P bond, was isolated from the fermn. broth
of Saccharothrix sp. ST-888. The biol. activity against various weeds
and total synthesis of phosphonothrixin are described herein.
Issue 3, 2000
Page 21
132: 35562f Stereospecific enzymatic hydrolysis of racemic

epoxide: a process for making chiral epoxide. Goswami, Animesh;
Totleben, Michael J.; Singh, Ambarish K.; Patel, Ramesh N. (Enzyme Technology, Process Research and Development, Bristol-Myers
Squibb Pharmaceutical Research Institute, New Brunswick, NJ 08903
USA). Tetrahedron: Asymmetry 1999, 10(16), 3167-3175 (Eng),
Elsevier Science Ltd.. Among various microbial cultures evaluated,
Rhodotorula glutinis SC 16293 and Aspergillus niger SC 16311
catalyzed the stereospecific hydrolysis of the racemic epoxide, RS-1{2',3'-dihydrobenzo[b]furan-4'-yl}-1,2-oxirane (1) to the corresponding R-diol, R-1-{2',3'-dihydrobenzo[b]furan-4'-yl}-ethane-1,2di ol (3). The S-epoxide, S-1-{2',3'-dihydrobenzo[b]furan-4'-yl}1,2-oxirane (2) remained unreacted in the reaction mixt. A reaction
yield of 45-50% (theor. max. yield is 50%) and an enantiomeric excess
(ee) of >95% were obtained for unreacted S-epoxide 2 using each
culture. Addn. of 10% Me tert-Bu ether to an aq. reaction mixt.
during hydrolysis by R. glutinis improved the ee of the unreacted
S-epoxide 2 to >99% (yield 48%) and that of the R-diol 3 to 79%.
Unlike R. glutinis, hydrolysis of racemic epoxide 1 in the presence of
10% Me tert-Bu ether by A. niger showed an adverse effect and gave
S-epoxide 2 in 54% yield and 49% ee.
132: 35970n Optical properties of dextran in solution and
films. Pavlov, G. M.; Grishchenko, A. E.; Ryumtsev, E. I.; Evlampieva, N. P.; Ivanitskii, V. V. (Institute of Physics, St. Petersburg
State University, St. Petersburg, Russia 199164). Biofizika 1999,
44(2), 251-256 (Russ), Nauka. A procedure for the quant. description of the simplest forms of self-organization of biol. mols. at
interfaces is proposed. The procedure uses the method of an inclined
polarized ray, which makes it possible to det. the degree of the orientational order of mol. fragments and the width of the anisotropic
surface layer. The quant. ests. were obtained in the study of the
microbial polysaccharide dextran using the methods of an inclined
polarized ray and flow birefringence. It was shown that mol. fragments of dextran are spontaneously oriented predominantly parallel
to the interface, and the thickness of oriented surface layers is
macroscopic (0.14 mm).
132: 35996a Preparation of non-nucleosidic coumarins as
polynucleotide-crosslinking agents. Wood, Michael L.; Cheng,
Peter C.; Thien, Douglas Y.; Albagli, David (Naxcor, USA) U.S. US
6,005,093 (Cl. 536-24.3; C07H21/04), 21 Dec 1999, US Appl. 46,568,
13 Apr 1993; 16 pp., Cont.-in-part of U.S. Ser. No. 46,568, abandoned. (Eng). Novel coumarin derivs. comprising a coumarin moiety
Y
Wn
linked to a non-nucleosidic backbone moiety I [B ) hydrocarbon,

heterocycle arom.; X ) bond, hydrocarbon, (un)substituted alkyl; W
) OH, halogen, amino, amido, azido, nitro, thio, carboxy, carbonyl,
perfluoromethyl, cyano, n ) 0-3; Y, Z ) H, F, alkyl] are disclosed.
The resulting mols. are typically used as photoactivate crosslinking
groups when incorporated into polynucleotides as replacements for
one or more of the complementary nucleoside bases present in probes
used in procedures involving nucleic acid hybridization reactions.
132: 37733m Microbial contamination of stored hydrocarbon
fuels and its control. Gaylarde, Christine C.; Bento, Fatima M.;
Kelley, Joan (Dep. Solos., Fac. Agron, Universidade Federal do Rio
Grande do Sul, CEP 91000-970 Porto Alegre, Brazil). Rev. Microbiol. 1999, 30(1), 1-10 (Eng), Sociedade Brasileira de Microbiologia.
A review, with 55 refs., discusses microbial contamination of gasoline,
aviation kerosene, and diesel fuel and its effects (e.g., quality loss,
sludge formation, and deterioration of pipes and storage tanks),
microorganisms isolated from hydrocarbon fuel systems, and contamination control using fuel additives, including biocides.
Page 22
NOTES
Issue 3, 2000

Microbial Biochemistry

Uploaded by

Document Information

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Microbial Biochemistry

Uploaded by

Copyright:

Available Formats

CA Selects: Fermentation Chemicals

antibiotic isolated from the fermn. broth of a Streptomyces sp. HIL

against oral pathogens. Groenink, J.; Walgreen-Weterings, E.;

been isolated as a major and a minor metabolite, resp., of Penicillium

themi normally produce radicinin and radicinol (I) when cultured on

132: 20869n Characterization of mutations in the rpoB gene

CA Selects: Fermentation Chemicals

these rickettsial species yielded topologies which were in accordance

toxic analogs of dolastatin G and nordolastatin G, resp., have been

after the cultivation of P. polymyxa up to its logarithmic growth phase,

CA Selects: Fermentation Chemicals

between intracellular pH and extreme acid resistance was low (R2<001).

CA Selects: Fermentation Chemicals

correlated with biosynthesis of VA. However, PAL induced by the

CA Selects: Fermentation Chemicals

132: 32957c Isolation of nitrogen-fixing bacteria containing

CA Selects: Fermentation Chemicals

lanostanoids, lucidadiol (I) and lucidal (II), were isolated from an

CA Selects: Fermentation Chemicals

ment IX is both essential and sufficient for the insertion of segment

CA Selects: Fermentation Chemicals

CA Selects: Fermentation Chemicals

which may be applied, as well as the limiting factors, possibly

procedures were objectively analyzed by an image anal. system. This

132: 33013d Control of Azospirillum brasilense NifA activity

132: 33010a Adaptive responses of Ralstonia eutropha to feast

132: 33014e Role of multiple efflux pumps in Escherichia coli

CA Selects: Fermentation Chemicals

effects of small perturbations in the host's existing pathways on the

CA Selects: Fermentation Chemicals

CA Selects: Fermentation Chemicals

132: 33041m Degradation of 2,4-dinitrophenol and selected

CA Selects: Fermentation Chemicals

Inc.; Cornell Research Foundation, Inc., USA) U.S. US 6,002,072

FERMENTATION & BIOINDUSTRIAL CHEMISTRY

patterns are shown to be engineered through a rational selection of

CA Selects: Fermentation Chemicals

2(2), No pp. Given (Eng), Universidad Catolica de Valparaiso. Avail.

132: 22197r Effect of harvesting protocol on performance of

CA Selects: Fermentation Chemicals

provide the xylose-metabolizing capability. In 1995, we achieved

CA Selects: Fermentation Chemicals

Park, J.-H.; Scott, A. I. (Department of Chemistry, Texas A&M

CA Selects: Fermentation Chemicals

Science Ireland Ltd.. Phenylacetaldehyde can be obtained by oxidn.

CA Selects: Fermentation Chemicals

Park, Jin-Byung; Han, Nam Soo; Ryu, Yeon-Woo; Seo, Jin-Ho

CA Selects: Fermentation Chemicals

are manufd. with Trichoderma. I and II are inhibitors to interleukin

CA Selects: Fermentation Chemicals

In comparison with the conventional solvent dewaxing process, the

CA Selects: Fermentation Chemicals

132: 35562f Stereospecific enzymatic hydrolysis of racemic

linked to a non-nucleosidic backbone moiety I [B ) hydrocarbon,

CA Selects: Fermentation Chemicals

You might also like