Campus Units
Biochemistry, Biophysics and Molecular Biology
Document Type
Article
Publication Version
Accepted Manuscript
Publication Date
4-2016
Journal or Book Title
New Phytologist
Volume
210
Issue
2
First Page
525
Last Page
534
DOI
10.1111/nph.13790
Abstract
Cytochromes P450 (CYPs) play key role in generating the structural diversity of terpenoids, the largest group of plant natural products. However, functional characterization of CYPs has been challenging because of the expansive families found in plant genomes, diverse reactivity and inaccessibility of their substrates and products.
• Here we present the characterization of two CYPs, CYP76AH3 and CYP76AK1, that act sequentially to form a bifurcating pathway for the biosynthesis of tanshinones, the oxygenated diterpenoids from the Chinese medicinal plant Danshen.
• These CYPs had similar transcription profiles to that of the known gene responsible for tanshinone production in elicited Danshen hairy roots. Biochemical and RNA interference studies demonstrated that both CYPs are promiscuous. CYP76AH3 oxidizes ferruginol at two different carbon centers, and CYP76AK1 hydroxylates C-20 of two of the resulting intermediates. Together, these convert ferruginol into 11,20-dihydroxy ferruginol and 11,20-dihydroxy sugiol en route to tanshinones. Moreover, we demonstrate the utility of these CYPs by engineering yeast for heterologous production of six oxygenated diterpenoids, which in turn enabled structural characterization of three novel compounds produced by CYP-mediated oxidation.
• Our results highlight the incorporation of multiple CYPs in diterpenoids metabolic engineering, and a continuing trend of CYPs promiscuity generating complex networks in terpenoid biosynthesis.
Copyright Owner
The Authors
Copyright Date
2015
Language
en
File Format
application/pdf
Recommended Citation
Guo, Juan; Ma, Xiaohui; Cai, Yuan; Ma, Ying; Zhan, Zhilai; Zhou, Yongjin J.; Liu, Wujun; Guan, Mengxin; Yang, Jian; Cui, Guanghong; Kang, Liping; Yang, Lei; Shen, Ye; Tang, Jinfu; Lin, Huixin; Ma, Xiaojing; Jin, Baolong; Liu, Zhenming; Peters, Reuben J.; Zhao, Zongbao K.; and Huang, Luqi, "Cytochrome P450 promiscuity leads to a bifurcating biosynthetic pathway for tanshinones" (2016). Biochemistry, Biophysics and Molecular Biology Publications. 125.
https://lib.dr.iastate.edu/bbmb_ag_pubs/125
Included in
Biochemistry, Biophysics, and Structural Biology Commons, Plant Breeding and Genetics Commons
Comments
This is the peer reviewed version of the following article: Guo, J., Ma, X., Cai, Y., Ma, Y., Zhan, Z., Zhou, Y. J., Liu, W., Guan, M., Yang, J., Cui, G., Kang, L., Yang, L., Shen, Y., Tang, J., Lin, H., Ma, X., Jin, B., Liu, Z., Peters, R. J., Zhao, Z. K. and Huang, L. (2016), Cytochrome P450 promiscuity leads to a bifurcating biosynthetic pathway for tanshinones. New Phytol, 210: 525–534, which has been published in final form at doi:10.1111/nph.13790. This article may be used for non-commercial purposes in accordance With Wiley Terms and Conditions for self-archiving.