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Identification of the sesquiterpene synthase AcTPS1 and high production of (–)-germacrene D in metabolically engineered Saccharomyces cerevisiae
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Identification of the sesquiterpene synthase AcTPS1 and high production of (–)-germacrene D in metabolically engineered Saccharomyces cerevisiae
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Identification of the sesquiterpene synthase AcTPS1 and high production of (–)-germacrene D in metabolically engineered Saccharomyces cerevisiae
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Journal Article
Identification of the sesquiterpene synthase AcTPS1 and high production of (–)-germacrene D in metabolically engineered Saccharomyces cerevisiae
2022
Overview
Background
The sesquiterpene germacrene D is a highly promising product due to its wide variety of insecticidal activities and ability to serve as a precursor for many other sesquiterpenes. Biosynthesis of high value compounds through genome mining for synthases and metabolic engineering of microbial factories, especially
Saccharomyces cerevisiae
, has been proven to be an effective strategy. However, there have been no studies on the de novo synthesis of germacrene D from carbon sources in microbes. Hence, the construction of the
S. cerevisiae
cell factory to achieve high production of germacrene D is highly desirable.
Results
We identified five putative sesquiterpene synthases (AcTPS1 to AcTPS5) from
Acremonium chrysogenum
and the major product of AcTPS1 characterized by in vivo, in vitro reaction and NMR detection was revealed to be (–)-germacrene D. After systematically comparing twenty-one germacrene D synthases, AcTPS1 was found to generate the highest amount of (–)-germacrene D and was integrated into the terpene precursor-enhancing yeast strain, achieving 376.2 mg/L of (–)-germacrene D. Iterative engineering was performed to improve the production of (–)-germacrene D, including increasing the copy numbers of
AcTPS1
,
tHMG1
and
ERG20
, and downregulating or knocking out other inhibitory factors (such as
erg9
,
rox1
,
dpp1
). Finally, the optimal strain LSc81 achieved 1.94 g/L (–)-germacrene D in shake-flask fermentation and 7.9 g/L (–)-germacrene D in a 5-L bioreactor, which is the highest reported (–)-germacrene D titer achieved to date.
Conclusion
We successfully achieved high production of (–)-germacrene D in
S. cerevisiae
through terpene synthase mining and metabolic engineering, providing an impressive example of microbial overproduction of high-value compounds.
Publisher
BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
Subject
/ Chemistry and Materials Science
/ Enzymes
/ Fungi
/ Genomes
/ Glucose
/ Identification and classification
/ Microbial Genetics and Genomics
/ Microbial synthesis of natural products originating from medicinal plants and fungi
/ NMR
/ Plasmids
/ Saccharomyces cerevisiae - genetics
/ Saccharomyces cerevisiae - metabolism
/ Sesquiterpenes, Germacrane - metabolism
/ Yeast
