In this study, the effects of deleting key genes of ethanol and glycerol biosynthesis pathway and branches of cytosolic Acetyl-CoA and farnesyl diphosphate on farnesene production was studied in Saccharomyces cerevisiae WHE4. With the utilization of CRISPR-cas9, eight engineered strains were obtained and cultivation was carried out at lab-scale fermentation. Compared with that of strain WHE4, results showed that the deletion of ethanol dehydrogenase (ADH3-6) had no effect on ethanol and farnesene production. The deletion of glycerol-3-phosphatase dehydrogenase (GPD1 or GPD2) caused the decline of glycerol production (decreased by 15% and 34%), the deletion of galactokinase (GAL1, GAL7 and GAL10) caused decreased transcriptional levels of the mevalonate pathway genes, and the deletion of them caused the decline of farnesene production. The farnesene production was increased by 29% with the deletion of geranylgeranyl diphosphate synthase (BTS1) and diacylglycerol pyrophosphate phosphatase (DPP1) genes. And the highest farnesene production of strain WHE4-33 (WHE4 Δbts1, Δdpp1) reached 1 578.91 mg/L at a 5 L bioreactor by fed-batch fermentation. In conclusion, the influence of deleting key genes of upper and downstream pathways of the mevalonate pathway on farnesene production was studied, and this would provide guidance for constructing effective terpenoid-producing platform in S. cerevisiae.
WANG Junhua
,
FU Wenwen
,
LI Youran
,
ZHU Huilin
,
XU Sha
,
SHI Guiyang
,
ZHANG Liang
,
DING Zhongyang
,
GU Zhenghua
. Influence of deleting key genes on farnesene production in Saccharomyces cerevisiae[J]. Food and Fermentation Industries, 2022
, 48(2)
: 1
-7
.
DOI: 10.13995/j.cnki.11-1802/ts.027491
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