Summary: “Improved production of fatty alcohols in cyanobacteria by metabolic

engineering”
By: Luis F. Ramos M. Iris

Due to its application as fuels, fatty alcohols have become of great industrial importance.
Even though these compounds can be commonly found in nature, biotechnology has
focused on the recombinant production of fatty alcohols.
Researchers like Wahlen and Hofvander have dedicated time and effort to characterize
enzymes responsible for the reduction of fatty aldehydes and fatty acyl-CoAs. Said enzyme
was later labeled as Maqu_2220, the first letter being an indicator of the organism where
the polypeptide was first obtained from: Marinobacter aquaeolei.
For the research of Yao, et al. a strain of Synechocystis (a cyanobacteria) was used to
express the maqu_2220 gene, regulated by PpetE and found alongside a spectinomycin
resistance gene. Since the plasmid used was named pFQ52, the new mutant strain was
labeled as Syn-FQ52. Furthermore, the researchers knocked out two genes, sllo208 and
sllo209, that coded for acyl-ACP reductases and aldehyde-deformylating oxyganse
respectively, which in theory would aid to the enhanced production of fatty alcohols. 2
different runs were conducted, the first only had sllo208 knocked out (Syn-FQ52D08)
while the second run had also sllo209 inactivated (Syn-FQ52D0809).
When comparing the knockout strains against the control and Syn-FQ52, neither Syn-
FQ52D08 and Syn-FQ52D0809 showed traces of hydrocarbon, indicating that the deletion
of sllo208 successfully redirected the metabolic pathway into fatty alcohol production. The
inhibition of hydrocarbon production would mean an accumulation of fatty aldehydes;
however, the researchers weren’t able to detect these compounds and attribute this
phenomenon to the action of a newly discovered aldehyde dehydrogenase found in
cyanobacteria.
An untransformed strain showed no production of fatty alcohols, while the Syn-FQ52 strain
was able to produce 0.21 mg/g Dry Weight (DW). Furthermore, Syn-FQ52D08 showed an
increase in the yield up to 0.60 mg/g of DW after 140h while Syn-FQ52D0809 showed a
bigger improvement, yielding 2.87 mg/ g DW after the same time period. Both knockout
strains had an enhanced yield due to the inhibition of hydrocarbon pathway; however, Syn-
FQ52D0809’s yield was further improved because the deletion of sllo209 inhibits the fatty
aldehyde biosynthesis pathway and increases the availability of substrate. Nevertheless, the
removal of genes sllo208 and sllo209 reduced slightly the growth rate of the knockout
strains.
The yields obtained behaved differently through time. Syn-FQ52 showed a continuous
increase of the yield while, on the contrary, Syn-FQ52D0809’s yield decreased throughout
time. Syn-FQ52D08 behaved differently, reaching a maximum yield (1.03 mg/g DW) at
274 before decreasing slightly at 384 h.
Even after these modifications, the amount of fatty alcohols produced by genetically
engineered cyanobacteria is not enough to compete against host cells like E. coli and yeasts.
Therefore, the researchers suggest optimizing synonymous codon usage to better match the
cyanobacteria and to introduce more metabolic flux to improve the yield.

References:
Yao, L., Qi, F., Tan, X. et al. Improved production of fatty alcohols in cyanobacteria by
metabolic engineering. Biotechnol Biofuels 7, 94 (2014).
https://doi.org/10.1186/1754-6834-7-94