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.
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Acknowledgments
We thank Prof. David R. Nelson for annotation of CYP716A155.
Funding
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).
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Huang, J., Zha, W., An, T. et al. Identification of RoCYP01 (CYP716A155) enables construction of engineered yeast for high-yield production of betulinic acid. Appl Microbiol Biotechnol 103, 7029–7039 (2019). https://doi.org/10.1007/s00253-019-10004-z
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DOI: https://doi.org/10.1007/s00253-019-10004-z