Food and Fermentation Industries >

2024 , Vol. 50 >Issue 20: 9 - 17

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.038171

Metabolic engineering of Saccharomyces cerevisiae for efficient synthesis of ursolic acid

  • ZHAO Xingying ,
  • ZENG Weizhu ,
  • ZHOU Jingwen
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  • 1(Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China)
    2(Science Center for Future Foods, Jiangnan University, Wuxi 214122, China)

Received date: 2023-12-05

  Revised date: 2024-01-10

  Online published: 2024-11-01

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Abstract

Ursolic acid, a pentacyclic triterpene compound with anti-inflammatory and antioxidant effects, has been widely applied in medicine, cosmetics, and various other fields.In this study, a “push-pull” metabolic engineering strategy was employed to increase the production of ursolic acid in Saccharomyces cerevisiae.Firstly, de novo biosynthesis of ursolic acid was achieved by heterologous expression and optimization of CADDS, CaCYP716A83, and HtCPR genes, resulting in 5.4 mg/L of ursolic acid.Subsequently, an expansion of the endoplasmic reticulum area was conducted to provide more adherent surface for membrane protease activity, obtaining 40.3 mg/L of ursolic acid.Due to multiple REDOX reactions was involved in the ursolic acid synthesis pathway, in which the NADPH was an essential cofactor.By enhancing the supply of NADPH, the ursolic acid titer was increased to 48.2 mg/L.Additionally, optimization of the electron transport chain and utilization of cytochrome b5 for efficient electron transfer to P450 enzyme further enhanced the electron supply, resulting in 74.6 mg/L of ursolic acid.Finally, by iteratively integrating these strategies including expanding endoplasmic reticulum area, enhancing NADPH availability, and electron transport efficiency and regulating the copy number of CaCYP716A83 and HtCPR genes, the ursoilc acid production was further enhanced to 84.3 mg/L.This study offers valuable insights for the development of efficient microbial cell factories for terpenoid synthesis, serving as a reference in the field.

Key words： ursolic acid; Saccharomyces cerevisiae; endoplasmic reticulum; NADPH; cytochrome b5

Cite this article

ZHAO Xingying , ZENG Weizhu , ZHOU Jingwen . Metabolic engineering of Saccharomyces cerevisiae for efficient synthesis of ursolic acid[J]. Food and Fermentation Industries, 2024 , 50(20) : 9 -17 . DOI: 10.13995/j.cnki.11-1802/ts.038171

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