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Culture Documents
By Sanket Lunawat
Microbial platform for alka(e)ne biosynthesis Many organisms, including plant, insect, bacteria,
yeast and fungus, have the ability to synthesize alka(e)nes. Some studies show that, alka(e)nes may provide
the host with additional ability to deal with harsh environ- ment or means of communication (e.g.
pheromones) [30,31,40,41]. Generally speaking, alka(e)ne yields in natural producers are usually low [42]. It is
unpractical to use these natural producers to generate alka(e)nes on industrial scale. However, a strain of
Aureobasidium mel- anogenum recently isolated from mangrove ecosystem in Hainan Province of China
shows an exception. This strain seems to be able to produce more than 28 g/L alkanes when provided with
optimal culture conditions [43_,44]. Given the fact that huge amount of microorgan- isms exist in nature, one
cannot rule out the possibility that some of them may have the potential to be used directly as a microbial
platform for alka(e)ne biosynthe-sis. As mentioned above, fatty acid metabolism is critical to the production of
alka(e)ne. Theoretically speaking, any microorganism with a highly effective fatty acid metabolic pathway has
the potential to become a microbial platform for alka(e)ne biosynthesis, even though they cannot produce
hydrocarbons naturally. Up to now, E. coli is the most extensively used microbialplatform for alka(e)ne
synthesis. The highest amount of alkane produced by microorganism is achieved in a meta-bolic engineered E.
coli (580.8 mg/L). Despite the alde-hyde reductase (>13) redundancy presence in E. coli, some studies show
that, when given fatty acids as sub-strate, E. coli can only assimilate a small amount (_24%) to generate
alka(e)nes and fatty alcohols, indicating its insufficient activity towards fatty acid utilization [45]. Therefore,
researchers tend to use glucose as carbon source for alka(e)ne generation, as well as engineer E. coli towards
high fatty acid accumulation at the same time. Employing yeast as host often results in lower yield than E. coli.
Evidence indicates that some yeast possess at least 12 alk genes which are responsible for alkane degradation
[46]. Alkane chain length between C9–11 also has poison- ous effect towards S. cerevisiae [47]. Moreover,
complex cellular structure of eukaryotes often hinders the meta- bolic process due to the
compartmentalization between cellular organelles [48]. Cofactors required for alka(e)ne generating enzymes
are often synthesized in mitochon- dria and peroxisome [49] whereas artificial routes from substrate to
alka(e)ne usually take place in cytosol. By repositioning the alka(e)ne biosynthesis pathway into the
peroxisome of S. cerevisiae, Zhou et al. enhanced the titer of alka(e)ne and fatty alcohols by seven-fold.
Additional deletion of certain genes increase the number of peroxi-some, leads to another three-fold titer
enhancement [15__]. Reports about alka(e)ne generating cyanobacteria can trace back to 1960’.
Cyanobacterias obtain their energy through photosynthesis and are able to use CO 2 as sole carbon source.
With sunlight and aeration, they can accumulate a lot of biomass in a short period of time [3]. With expression
of multiple copies of AAR and ADO, metabolic engineered cyanobacteria can produce alkanes up to 12.9% of
their cell dry weight [50_,51,52]. Metabolic engineered chemoautotrophic bacteria Cupriavidus neca-tor can
also transform CO2 into alka(e)nes [8__]. Due to the abundance of CO2 in the atmosphere and its special role as
a greenhouse gas, CO2-assimilating microbes like cyanobacteria or C. necator represent a unique type of
microbial alka(e)ne producing platform which has a very promising future.
Conclusions
Among renewable biofuels, the most commonly used in todays market are bioethanol and biodiesel. Wild
type S. cerevisiae KL17 can produce ethanol at a concentration of 96.9 g/L [62]. Although biodiesel is
mainly produced by chemical transesterification of plant oil with alcohol, in situ esterification can also
lead to a biodiesel titer of 1.28 g/ L by coupling microbial fatty acid assimilation with ethanol formation in
metabolic engineered E. coli [63]. However, compared to bioethanol and biodiesel, low yield of microbial
produced alka(e)ne still has a long way to go to fulfill industrial use. Continuing search for efficient
alka(e)ne biosynthesis pathway is an essential task as well as the search for suitable microbial platform.
Most work have taken place in E. coli, which lacks the strong ability in fatty acid accumulation and
assimilation even with intensive genetic modifications. We believe it is beneficial to employ different
kinds of oleaginous, lipolytic and CO2-fixing microbes into the production of alka(e)nes in future. These
microorganisms will bestow the system with higher efficiency, shorter metabolic pathway and cheaper
feedstock. Successful integration of these microbes with efficient alkane generating path-way should
deliver more satisfactory alka(e)ne yield. Under optimized culture conditions, these alka(e)ne gen-erating
platform may one day fulfill industrial purposes. Conflicts of interest None. .