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Applied Microbiology and Biotechnology

, Volume 103, Issue 2, pp 761–776 | Cite as

Regio- and enantioselective O-demethylation of tetrahydroprotoberberines by cytochrome P450 enzyme system from Streptomyces griseus ATCC 13273

  • Chen Shen
  • Tianyue Shan
  • Wanli Zhao
  • Chenhui Ou
  • Li Li
  • Xiufeng LiuEmail author
  • Jihua LiuEmail author
  • Boyang Yu
Biotechnologically relevant enzymes and proteins

Abstract

Tetrahydroprotoberberines (THPBs), a class of naturally occurring isoquinoline alkaloids, contain substituent methoxyl or hydroxyl groups which play a significant role in the pharmacological properties of these molecules. In this study, we report a biocatalytic strategy for selective O-demethylation of THPBs. CYP105D1, a cytochrome P450 from Streptomyces griseus ATCC 13273, exhibited markedly regioselective demethylation of nonhydroxyl-THPBs and monohydroxyl-THPBs on the D-ring. A possible binding mode of THPBs with CYP105D1 was investigated by docking analysis, and the results revealed that the D-rings of THPBs were with the minimum distance to the heme iron. Tetrahydropalmatine was used as a model substrate and enantioselective demethylation was demonstrated. (S)-Tetrahydropalmatine was only demethylated at C-10, while (R)-tetrahydropalmatine was first demethylated at C-10 and then subsequently demethylated at C-9. The kcat/Km value for demethylation of (R)-tetrahydropalmatine by CYP105D1 was 3.7 times greater than that for demethylation of (S)-tetrahydropalmatine. Furthermore, selective demethylation of (S)-tetrahydropalmatine by the CYP105D1-based whole-cell system was demonstrated for the highly efficient production of (S)-corydalmine which has distinct pharmacological applications, such as providing relief from bone cancer pain and reducing morphine tolerance. Moreover, a homologous redox partner was identified to enhance the catalytic efficiency of the CYP105D1-based whole-cell system. This is the first enzymatic characterization of a cytochrome P450 that has regio- and enantioselective demethylation activity of THPBs for application purpose. The cytochrome P450 system could be a promising strategy for selective demethylation in the pharmaceutical industry.

Keywords

Biotransformation Corydalmine Cytochrome P450 Demethylation Redox partner Tetrahydroprotoberberine 

Notes

Funding information

This work was supported by the Major Scientific and Technological Specialized Project for “New Drugs Development” (No. 2012ZX09J12110-06B) and the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

Compliance with ethical standards

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

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

253_2018_9416_MOESM1_ESM.pdf (1.2 mb)
ESM 1 (PDF 1211 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Jiangsu Key Laboratory of TCM Evaluation and Translational Research, School of Traditional Chinese PharmacyChina Pharmaceutical UniversityNanjingChina
  2. 2.State Key Laboratory of Natural MedicinesChina Pharmaceutical UniversityNanjingChina

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