Structure-function analysis of CYP719As involved in methylenedioxy bridge-formation in the biosynthesis of benzylisoquinoline alkaloids and its de novo production
Artikel i vetenskaplig tidskrift, 2023

Benzylisoquinoline alkaloids (BIAs) are a type of secondary metabolite with clinical application value. (S)-stylopine is a special BIA which contains methylenedioxy bridge structures. CYP719As could catalyze the methylenedioxy bridge-formation on the A or D rings of protoberberine alkaloids, while displaying significant substrate regiospecificity. To explore the substrate preference of CYP719As, we cloned and identified five CyCYP719A candidates from Corydalis yanhusuo. Two CyCYP719As (CyCYP719A39 and CyCYP719A42) with high catalytic efficiency for the methylenedioxy bridge-formation on the D or A rings were characterized, respectively. The residues (Leu 294 for CyCYP719A42 and Asp 289 for CyCYP719A39) were identified as the key to controlling the regioselectivity of CYP719As affecting the methylenedioxy bridge-formation on the A or D rings by homology modeling and mutation analysis. Furthermore, for de novo production of BIAs, CyCYP719A39, CyCYP719A42, and their mutants were introduced into the (S)-scoulerine-producing yeast to produce 32 mg/L (S)-stylopine. These results lay a foundation for understanding the structure-function relationship of CYP719A-mediated methylenedioxy bridge-formation and provide yeast strains for the BIAs production by synthetic biology.

Methylenedioxy bridge-formation

Regiospecificity

Corydalis yanhusuo

Synthetic biology

Benzylisoquinoline alkaloids (BIAs)

CyCYP719As

Författare

Xiuyu Liu

Henan University of Chinese Medicine

China Academy of Chinese Medical Sciences

Xiang Jiao

Chalmers, Life sciences, Systembiologi

Yatian Cheng

China Academy of Chinese Medical Sciences

Ying Ma

China Academy of Chinese Medical Sciences

Junling Bu

China Academy of Chinese Medical Sciences

Baolong Jin

China Academy of Chinese Medical Sciences

Qishuang Li

China Academy of Chinese Medical Sciences

Zhimin Hu

China Academy of Chinese Medical Sciences

Jinfu Tang

China Academy of Chinese Medical Sciences

Changjiangsheng Lai

China Academy of Chinese Medical Sciences

Jian Wang

China Academy of Chinese Medical Sciences

Guanghong Cui

China Academy of Chinese Medical Sciences

Yun Chen

Chalmers, Life sciences, Systembiologi

Juan Guo

China Academy of Chinese Medical Sciences

Luqi Huang

China Academy of Chinese Medical Sciences

Microbial Cell Factories

14752859 (eISSN)

Vol. 22 1 23

Ämneskategorier

Biokemi och molekylärbiologi

Biokatalys och enzymteknik

Organisk kemi

DOI

10.1186/s12934-023-02024-2

PubMed

36737755

Mer information

Senast uppdaterat

2023-02-20