期刊
ENZYME AND MICROBIAL TECHNOLOGY
卷 134, 期 -, 页码 -出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.enzmictec.2019.109462
关键词
Linalool; Saccharomyces cerevisiae; ERG20 mutant; MVA pathway; Pathway compartmentation
资金
- National 948 Project of China [2014-4-33]
- National Key Research and Development Program of China [2016YFD0600801]
Linalool, a valuable monoterpene alcohol, is widely used in cosmetics and flavoring ingredients. However, its scalable production by microbial fermentation is not yet achieved. In this work, considerable increase in linalool production was obtained in Saccharomyces cerevisiae by dual metabolic engineering of the mevalonic acid (MVA) pathway in both mitochondria and cytoplasm. A farnesyl pyrophosphate synthase mutant ERG20(F96W/N127W) and a linalool synthase from Cinnamomum osmophloeum (CoLIS) were introduced and meanwhile the endogenous ERG20 was down-regulated to prevent the competitive loss of precursor. In addition, overexpression of the proteins of CoLIS and ERG20(F96W/N127W) and another copy of the same enzymes CoLIS/ERG20(F96W/N127W) with mitochondrial localization signal (MLS) were carried out to further pull the flux to linalool. Finally, a maximum linalool titer of 23.45 mg/L was obtained in a batch fermentation with sucrose as carbon source. This combinatorial engineering strategy may provide hints for biosynthesis of other monoterpenes.
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