Cloning and Function of Gene for Cytochrome P450 from a Chlorogenic Acid-producing Endophytic Bacterium

doi:10.13560/j.cnki.biotech.bull.1985.2016.06.026

Abstract

Abstract: Based on a chlorogenic acid-producing endophytic Bacillus subtilis，cloning and function of a key gene in the synthetic pathway of chlorogenic acid was studied. The cytochrome P450 gene of the endophyte was cloned and expressed in prokaryotic vector，and the cinnamate 4-hydroxylase(C4H)activity of the recombinant proteins were determined. Four cytochrome P450 genes were cloned and expressed in prokaryotic vector，and the recombinant protein P-4 presented C4H activity. The coumarate produced by the C4H activity was the same as the result measured by LC-MS. The activity of P-4 existed in a broad optimal temperature and was inhibited by product coumarate. It is supposed that，taking cytochrome P450 as isozyme of C4H，the endophyte led the product coumarate to the synthetic pathway of chlorogenic acid.

Key words: chlorogenic acid, cytochrome P450, C4H, expression, activity

WANG Chuan, LI Li, WEI Pi-wei, HUANG Fei. Cloning and Function of Gene for Cytochrome P450 from a Chlorogenic Acid-producing Endophytic Bacterium[J]. Biotechnology Bulletin, 2016, 32(6): 181-186.

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