Special Abstracts / Journal of Biotechnology 150S (2010) S1S576 S19

[B.28]
Cloningandbioinformatic characterizationof genes controlling
key steps of the fatty acid biosynthesis and lipid breakdown in
seeds of Jatropha curcas L.
D.G. Ambrosi, G. Galla, S. Collani, G. Barcaccia

Laboratory of Plant Genetics and Genomics, University of Padua, Italy

Keywords: Jatropha curcas; Gene cloning; Lipid metabolism; Bio-
fuel
Jatropha curcas L. (2n=2x =22) is becoming a popular non-food
oleaginous crop in several developed countries for its proposed
value in the biopharmaceutical industry. Despite the potentials
of its oil-rich seeds as a renewable source of biodiesel and
an interest in large-scale cultivation, relatively little is known
with respect to population genetics and plant genomics. We
recently performed genomic DNA markers and FCSS analyses
to gain insights on ploidy variation and heterozygosity lev-
els of multiple accessions, and genomic relationships among
commercial varieties and locally dominant ecotypes grown in
different geographical areas. Seeds commercialized worldwide
seem to include a few closely related genotypes showing high
degrees of homozygosity for single varieties and very low
genetic diversity between varieties. For a better understand-
ing of oil production and accumulation in J. curcas seeds, it
would be useful to clone and characterize genes controlling key
steps of FA biosynthesis and lipid breakdown pathways. Gene
bank searches and bioinformatic analyses allowed us to select
13 genes encoding for enzymes involved in lipid metabolism.
Five of these genes were previously isolated in J. curcas (i.e.,
acetyl-CoA carboxylase I, 3-ketoacyl-ACP I and III, acyl-ACP desat-
urase, and acyl-ACP thioesterase II), and their nucleotide and
amino acid sequences are available in the NCBI databases. The
remaining eight genes were cloned in the present study (i.e.,
ATP-citrate lyase, acetyl-CoA carboxylase II, 3-ketoacyl-ACP II, 3-
ketoacyl-ACP reductase, 3-hydroxyacyl-ACP dehydrase, enoyl-ACP
reductase, acyl-ACP thioesterase I, and acyl-CoA dehydrogenase).
Replicated cDNA samples were produced by retrotranscription
of mRNAs isolated from mature seeds belonging to different
commercial varieties and Mexican ecotypes. Gene expression
levels were assayed by quantitative Real-Time PCR with gene-
specic primer combinations. The haplotyping of single accessions
will be attempted using a worldwide collection of J. curcas
materials to nd out the most representative and discrimi-
nant SNPs related to FA biosynthesis and lipid metabolism
genes.
doi:10.1016/j.jbiotec.2010.08.061
[B.29]
Development of innovative technologies for the optimal cultur-
ing and characterization of microalgae as energy biomass
G. Carpani
1,2
, F. Capuano
1,2
, E. Daddario
1,2
, F. de Ferra
1,2,
1
R&M Division - Downstream Process Technology (SDM-PROC), Italy
2
Energy Env Processes and Technologies (PTEA), Italy
Microalgae represent a potentially interesting type of biomass
tobe usedas a source of molecules that canbe renedandupgraded
to biofuels. In fact the biological characteristics of growth and lipid
accumulation make microalgal strains different enough fromother
vegetal species tooffer newprospective scenarios of exploitationas
energy biomass incomparisonto other plants andmicroorganisms.
Still signicant scientic and technological issues at the basis
of biomass productivity and economic fuel production have to be
studied and solved to achieve efcient processes and potentially
economical production cycles.
Pilot sized experimental apparati located at a renery site allow
recent experimentation at a scale necessary to evaluate some of
the fundamental issues involved in the assessment of industrial
applicability and to rene the colturing and work up technologies
involved in the biomass to fuel production.
In this occasion recent results will be presented on the opti-
mization of culturing processes, and the development of molecular
methods for the characterization of pond-grown microalgal con-
sortia.
doi:10.1016/j.jbiotec.2010.08.062
[B.30]
Inuences of CO
2
concentrations and salinity on acceleration of
microalgal oil as raw material for biodiesel production
Prayoon Enmak
1,2,
, Pakawadee Kaewkannetra
2,3
1
School of Khon Kaen University, Khon Kaen 40002, Thailand
2
Centre for Alternative Energy Research and Development (AERD),
Faculty of Engineering, Khon Kaen University, Khon Kaen 40002, Thai-
land
3
Department of Biotechnology, Faculty of Technology, Khon Kaen Uni-
versity, Khon Kaen 40002, Thailand
Keywords: CO
2
; Salinity; Microalgal oil; Scenedesmus obliquus;
Biodiesel
In this work, several microalgae samples were collected
from natural water basins in northern area of Thailand using
micropipette technique. The isolationstep was investigated by reg-
ular observation under microscope technique. The algal strains
were enriched in modied Chu 13 medium. The morphology of the
algal isolated strain showed in a green unicellular colony thereafter
as identiedas Scenedesmus obliquus. Then, growthmonitoring was
performed in 3L vertical tubular glass photo-reactors containing
modied Chu 13 medium under autotrophic cultivations (16:8h
of light dark cycle using cool white uorescent at room temper-
ature) coupling which injection of different CO
2
concentrations
(5, 10 and 15%) mixed with ambient air for achieving the maxi-
mum algal biomass. The results revealed that the S. Obliquus can
be grown in all variations of CO
2
concentrations. Moreover, it was
found that the algal biomass yields increased as CO
2
concentra-
tions increase. The maximum biomass was obtained at 2.18 gL
1
(dry cell weight) when 15% CO
2
concentration was used. Hereafter,
the effect of salinity on the oil accumulation in the S. Obliquus was
considered. Sodium chloride (NaCl) solutions (0.1, 0.2 and 0.3M)
were prepared with deionized water and then were added into the
cultures when the algal cells reached at early stationary phase (salt
stress period). The algal biomass was harvested at day 5th, 10th
and 15th after salt stress. Biodiesel was performed by in-situ acidic
transesterication. The biodiesel property obtained was subjected
to American Society for Testing and Materials Standard (ASTM),
such as ash point, pH, density and viscosity.
doi:10.1016/j.jbiotec.2010.08.063