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Applied Microbiology and Biotechnology

, Volume 103, Issue 17, pp 7029–7039 | Cite as

Identification of RoCYP01 (CYP716A155) enables construction of engineered yeast for high-yield production of betulinic acid

  • Jiajian Huang
  • Wenlong Zha
  • Tianyue An
  • Hua Dong
  • Ying Huang
  • Dong Wang
  • Rongmin Yu
  • Lixin Duan
  • Xueli Zhang
  • Reuben J. PetersEmail author
  • Zhubo DaiEmail author
  • Jiachen ZiEmail author
Biotechnological products and process engineering

Abstract

Betulinic acid (BA) and its derivatives possess potent pharmacological activity against cancer and HIV. As with many phytochemicals, access to BA is limited by the requirement for laborious extraction from plant biomass where it is found in low amounts. This might be alleviated by metabolically engineering production of BA into an industrially relevant microbe such as Saccharomyces cerevisiae (yeast), which requires complete elucidation of the corresponding biosynthetic pathway. However, while cytochrome P450 enzymes (CYPs) that can oxidize lupeol into BA have been previously identified from the CYP716A subfamily, these generally do not seem to be specific to such biosynthesis and, in any case, have not been shown to enable high-yielding metabolic engineering. Here RoCYP01 (CYP716A155) was identified from the BA-producing plant Rosmarinus officinalis (rosemary) and demonstrated to effectively convert lupeol into BA, with strong correlation of its expression and BA accumulation. This was further utilized to construct a yeast strain that yields > 1 g/L of BA, providing a viable route for biotechnological production of this valuable triterpenoid.

Keywords

Betulinic acid Cytochrome P450 Synthetic biology Yeast 

Notes

Acknowledgments

We thank Prof. David R. Nelson for annotation of CYP716A155.

Funding information

This work was supported by grants from the National Natural Science Foundation of China (NSFC, No. 81673530), China Postdoctoral Science Foundation (2018 M633289), and Open Fund of Guangdong Province Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs Research (No. 2016B 030301004).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2019_10004_MOESM1_ESM.pdf (456 kb)
ESM 1 (PDF 455 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Biotechnological Institute of Chinese Materia MedicJinan UniversityGuangzhouChina
  2. 2.Tianjin Institute of Industrial BiotechnologyChinese Academy of SciencesTianjinChina
  3. 3.Guangdong Province Key Laboratory of Pharmacodynamic Constituents of Traditional Chinese Medicine and New Drugs ResearchJinan UniversityGuangzhouChina
  4. 4.Joint Laboratory for Translational Cancer Research of Chinese Medicine of the Ministry of Education of the People’s Republic of China, International Institute for Translational Chinese MedicineGuangzhou University of Chinese MedicineGuangzhouChina
  5. 5.Roy J. Carver Department of Biochemistry, Biophysics, Molecular BiologyIowa State UniversityAmesUSA

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