Identification and characterization of methyltransferases involved in benzylisoquinoline alkaloids biosynthesis from Stephania intermedia

Objectives To characterize methyltransferases involved in the biosynthesis of benzylisoquinoline alkaloids in Stephania intermedia . Results Three N -methyltransferases, SiCNMT1, SiCNMT2, SiCNMT3, and O -methyltransferase SiSOMT were identified in Stephania intermedia . Then, four methyltransferase...

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Published in:Biotechnology letters 2020-03, Vol.42 (3), p.461-469
Main Authors: Zhao, Wanli, Shen, Chen, Zhu, Jinqian, Ou, Chenhui, Liu, Manyu, Dai, Wenling, Liu, Xiufeng, Liu, Jihua
Format: Article
Language:English
Subjects:
Alkaloids
Applied Microbiology
Biochemistry
Biomedical and Life Sciences
Biosynthesis
Biotechnology
E coli
Expression vectors
Life Sciences
Methylation
Methyltransferase
Microbiology
Original Research Paper
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Summary:Objectives To characterize methyltransferases involved in the biosynthesis of benzylisoquinoline alkaloids in Stephania intermedia . Results Three N -methyltransferases, SiCNMT1, SiCNMT2, SiCNMT3, and O -methyltransferase SiSOMT were identified in Stephania intermedia . Then, four methyltransferase genes were cloned into the pGEX-6P-1 vector. The recombinant vectors were transformed into Escherichia coli BL21(DE3) for expression and were functionally tested. SiCNMT1, SiCNMT2, and SiCNMT3 could methylate ( R )-coclaurine to produce ( R )- N -methylcoclaurine. SiCNMT2 further methylated the product of ( R )- N -methylcoclaurine to produce ( R )-magnocurarine. Similarly, ( R )-norcoclaurine was continuously catalyzed to yield ( R )- N -methylnorcoclaurine and ( R )- N, N -dimethylnorcoclaurine by SiCNMT2. Furthermore, SiSOMT was shown to catalyze the conversion of ( S )-scoulerine to ( S )-tetrahydropalmatine. Conclusions The key methyltransferases, which were in the last step biosynthesis of ( R )-magnocurarine, ( R )- N , N -dimethylnorcoclaurine and ( S )-tetrahydropalmatine were revealed and their activities were verified in vitro. Four novel methyltransferases will be promising candidates for methylation of benzylisoquinoline alkaloids.
ISSN:0141-5492
1573-6776
DOI:10.1007/s10529-019-02785-0