Rate-limiting steps in the Saccharomyces cerevisiae ergosterol pathway: towards improved ergosta-5,7-dien-3β-ol accumulation by metabolic engineering

  • Bin-Xiang Ma
  • Xia Ke
  • Xiao-Ling Tang
  • Ren-Chao Zheng
  • Yu-Guo Zheng
Original Paper

Abstract

Ergosterol is the predominant nature sterol constituent of plasma membrane in Saccharomyces cerevisiae. Herein, the biosynthetic pathway of ergosterol was proposed to be metabolically engineered for the efficient production of ergosta-5,7-dien-3β-ol, which is the precursor of vitamin D4. By target disruption of erg5, involved in the end-steps of post-squalene formation, predominantly accumulated ergosta-5,7-dien-3β-ol (4.12 mg/g dry cell weight). Moreover, the rate-limiting enzymes of ergosta-5,7-dien-3β-ol biosynthesis were characterized. Overexpression of Hmg1p led to a significant accumulation of squalene, and induction of Erg1p/Erg11p expression raised the yield of both total sterols and ergosta-5,7-dien-3β-ol with no obvious changes in growth behavior. Furthermore, the transcription factor allele upc2-1 was overexpressed to explore the effect of combined induction of rate-limiting enzymes. Compared with an obviously enhanced yield of ergosterol in the wild-type strain, decreases of both the ergosta-5,7-dienol levels and the total sterol yield were found in Δerg5-upc2-1, probably due to the unbalanced NADH/NAD+ ratio observed in the erg5 knockouts, suggesting the whole-cell redox homeostasis was also vital for end-product biosynthesis. The data obtained in this study can be used as reference values for the production of sterol-related intermediates involved in the post-squalene biosynthetic pathway in food-grade S. cerevisiae strains.

Graphical Abstract

Keywords

Ergosta-5,7-dien-3β-ol Ergosterol synthesis pathway Rate-limiting steps Saccharomyces cerevisiae Upc2-1 

Notes

Acknowledgements

This work was supported by the National Natural Science Foundation of China (No. 31400978) and the Natural Science Foundation of Zhejiang Province (No. LY18B020019).

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

11274_2018_2440_MOESM1_ESM.doc (41 kb)
Supplementary material 1 (DOC 41 KB)

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Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  • Bin-Xiang Ma
    • 1
    • 2
  • Xia Ke
    • 1
    • 2
  • Xiao-Ling Tang
    • 1
    • 2
  • Ren-Chao Zheng
    • 1
    • 2
  • Yu-Guo Zheng
    • 1
    • 2
  1. 1.Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and BioengineeringZhejiang University of TechnologyHangzhouPeople’s Republic of China
  2. 2.Engineering Research Center of Bioconversion and Biopurification of Ministry of EducationZhejiang University of TechnologyHangzhouPeople’s Republic of China

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