CULTURE MEDIUM FOR Bacillus sphaericus
A gene expression cassette comprising
Kalia VC and Purohit HJ, Oklahoma City, Okla
No Drawing, Application June 21, 2002
Serial No. 354,544
8 Claims,
To provide a new microorganism capable
of producing a polyhydroxyalkanoate (PHA), a
PHA synthase gene, an expression cassette
including the gene, a vector including the
expression cassette, a transformant transformed
by the vector, a polypeptide having PHA
synthase activity, a method for producing a PHA
synthase and a method for producing a PHA.
Solution: The new microorganism is capable of
producing a polyhydroxyalkanoate comprising a
16S rRNA gene whose polynucleotide sequence
shows 99% or more homology to a
polynucleotide sequence represented by SEQ ID
No: 1, having an optimum temperature of an
activity temperature range for the growth and
PHA production of the microorganism of at least
34° C. and being capable of growing at a pH
range from 6 to 10.
Claims
1 A microorganism being capable of producing a
polyhydroxyalkanoate (PHA) and comprising a
16S rRNA gene (16S rDNA) whose
polynucleotide sequence shows 99% or more
homology to a polynucleotide sequence
represented by SEQ ID No: 1, comprising the
following characteristics (a) and (b):(a) having
an optimum temperature of an activity
temperature range for the growth and PHA
production of said microorganism of at least
34.degree. C.;(b) being capable of growing at a
pH range from 6 to 10.
2. A microorganism being capable of
producing a polyhydroxyalkanoate (PHA) and
being Pseudomonas sp. SG4502 (NITE BP-
578).
3.
said PHA synthase I gene as set forth in claim 3,
said PHA synthase II gene as set forth in claim
4. 4. and one or more genes selected from
the group consisting of genes of
polyhydroxyalkanoate-degrading enzymes,
wherein PHA-degrading enzyme(i) encodes a
polypeptide having at least one of the following
characteristic (a), (b) and (c):(a) a polypeptide
comprising an amino acid sequence represented
by SEQ ID No: 7;(b) a polypeptide having PHA
degrading activity and comprising an amino
acid sequence, in which one or more amino
acids are deleted, substituted, inserted and/or
added in said polypeptide comprising an amino
acid sequence represented by SEQ ID No: 7;(c)
a polypeptide having at least 80% or more of
amino acid sequence identity with respect to
said polypeptide comprising an amino acid
sequence represented by SEQ ID No: 7 and
PHA degrading activity
5. Culture Medium were prepared as follows:
Carbon Source (Glucose, Sucrose or Molsses)
Each liter of culture medium consist of
bacillus sphaericus depend of carbon source
used... Research about polyhydroxyvalerat has
been developed since PHA produced as a
commercial products.
Any research founds the composition available
for each carbon source:
Carbon Source Equivalent
Composition
Molsses 9 / 20 v/v%
Glucose 7/ 20 v/v%
Sucrose 7 / 20 v/v%
Nutrient Limit
Using difference nutrient limit should
be produce the difference
polyhydroxyalcanoates chain. It will be
understood that the proportions of the various
nutrient components of the culture medium may
be varied considerably and may be adjusted to 6. Anil-Kumar PK, Shamla TR, Kshama L,
the optimum requirements of the specisic Prakash MH, Joshi GJ, Chandrashekar A,
organisms. Kumari KSL, Divyashree MS: Bacterial
synthesis of poly(hydroxybutyrate-co-
6. A transformant as set forth in claim 4 being hydroxyvalerate) using carbohydrate-rich
mahua (Madhuca sp.) flowers. J Appl
Bacillus sphaericus
Microbiol 2007, 103:204-209.
7. Wu Q, Huang H, Hu G, Chen J, Ho KP,
7. A method for producing a Chen GQ: Production of poly-3-
polyhydroxyalkanoate (PHA) synthase hydroxybutyrate by Bacillus sp. JMa5
comprising collecting a polyhydroxyalkanoate cultivated in molasses media. Antonie van
(PHA) synthase from a culture obtained by Leeuwenhoek 2001, 80:111-118.
culturing said microorganism as set forth in 8. Vijayendra SVN, Rastogi NK, Shamala TR,
claim 1 or 2. Anil-Kumar PK, Kshama L, Joshi GJ:
Optimization of polyhydroxybutyrate
8. A method for producing a production by Bacillus sp CFR 256 with corn
polyhydroxyalkanoate (PHA) comprising steep liquor as a nitrogen source. Ind J
collecting a polyhydroxyalkanoate (PHA) from Microbiol 2007, 47:170-175.
a culture obtained by culturing said 9. Wang YJ, Hua FL, Tseng YF, Chan SY, Sin
microorganism as set forth in claim 1 or 2. SN, Chua H, Yu PHF, Ren NQ: Synthesis of
PHAs from waste water under various C:N
ratios. Biores Technol 2007, 98:1690-1693.
***

References

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