Construction of Biocatalysts Using the P450 Scaffold for the Synthesis of Indigo from Indole

文献类型： 外文期刊

第一作者： Li, Yanqing

作者： Li, Yanqing;Li, Yanqing;Wang, Fang;Wang, Jinghan;Xu, Jiakun;Lin, Yingwu;Wang, Jinghan;Shoji, Osami

作者机构：

关键词： cytochromes P450BM3; dual-functional small molecule co-catalysis; hydroxylated indole; indigo; directed evolution

期刊名称：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES （ 影响因子：5.6； 五年影响因子：6.2 ）

ISSN：

年卷期： 2023 年 24 卷 3 期

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收录情况： SCI

摘要： With the increasing demand for blue dyes, it is of vital importance to develop a green and efficient biocatalyst to produce indigo. This study constructed a hydrogen peroxide-dependent catalytic system for the direct conversion of indole to indigo using P450BM3 with the assistance of dual-functional small molecules (DFSM). The arrangements of amino acids at 78, 87, and 268 positions influenced the catalytic activity. F87G/T268V mutant gave the highest catalytic activity with k(cat) of 1402 min(-1) and with a yield of 73%. F87A/T268V mutant was found to produce the indigo product with chemoselectivity as high as 80%. Moreover, F87G/T268A mutant was found to efficiently catalyze indole oxidation with higher activity (k(cat)/K-m = 1388 mM(-1) min(-1)) than other enzymes, such as the NADPH-dependent P450BM3 (2.4-fold), the Ngb (32-fold) and the Mb (117-fold). Computer simulation results indicate that the arrangements of amino acid residues in the active site can significantly affect the catalytic activity of the protein. The DFSM-facilitated P450BM3 peroxygenase system provides an alternative, simple approach for a key step in the bioproduction of indigo.

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