4.7 Article

Combinatorial Modulation of Linalool Synthase and Farnesyl Diphosphate Synthase for Linalool Overproduction in Saccharomyces cerevisiae

期刊

JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
卷 69, 期 3, 页码 1003-1010

出版社

AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.0c06384

关键词

linalool; Saccharomyces cerevisiae; geranyl pyrophosphate; linalool synthase; farnesyl diphosphate synthase

资金

  1. National Natural Science Foundation of China [32001032]
  2. Natural Science Foundation of Jiangsu Province, China [BK20200946]
  3. China Postdoctoral Science Foundation [2020M671614]
  4. Natural Science Foundation of the Jiangsu Higher Education Institutions of China [20KJD416003]
  5. Innovative and Entrepreneurial Talent of Jiangsu Province
  6. Lvyang Jinfeng Talent Attracting Plan of Yangzhou

向作者/读者索取更多资源

By combinational regulation of linalool synthase and farnesyl diphosphate synthase, a stress-free linalool-producing yeast cell factory was constructed, achieving the highest reported linalool production in yeast and showing potential for producing other high-value monoterpenes.
Linalool, as a fragrant monoterpene, is an important feedstock for food, pharmaceuticals, and cosmetics industries. Although our previous study had significantly increased linalool production by the directed evolution of linalool synthase and overexpression of the whole mevalonate pathway genes, the engineered yeast strain suffered from dramatically reduced biomass. Herein, a stress-free linalool-producing yeast cell factory was constructed by the combinational regulation of linalool synthase and farnesyl diphosphate synthase instead of multienzyme overexpression. First, the expression level of linalool synthase was successfully enhanced by introducing a N-terminal SKIK tag, which improved linalool production by 3.3-fold. Subsequently, the modular assembly of linalool synthase and dominant negative farnesyl diphosphate synthase via short peptide tags efficiently converted geranyl pyrophosphate to linalool. Additional downregulation of the native farnesyl diphosphate synthase led to the highest reported linalool production (80.9 mg/L) in yeast. This combinatorial modulation strategy may also be applied to the production of other high-value monoterpenes.

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