文献类型：期刊文献

中文题名：非特异性过加氧酶(UPO)的研究综述

英文题名：Review of research on unspecific peroxygenases(UPOs)

作者：赖铭元[1];韦健[1];许建和[1];郁惠蕾[1]

机构：[1]华东理工大学,生物反应器工程国家重点实验室,上海200237

年份：2022

卷号：3

期号：6

起止页码：1235

中文期刊名：合成生物学

外文期刊名：Synthetic Biology Journal

收录：CSCD:【CSCD_E2021_2022】；

基金：国家重点研发计划(2019YFA0905000)。

语种：中文

中文关键词：非特异性过加氧酶;未活化C—H键;氧化反应;异源表达;选择性

外文关键词：unspecific peroxygenase(UPO);unactivated C—H bond;oxidation reaction;heterologous expression;selectivity

摘要：未活化的C—H键选择性插入活性氧是目前有机合成面临的最具挑战性之一。真菌非特异性过加氧酶(UPO)是一类高度糖基化的硫代血红素酶,催化的反应包括正构烷烃中未活化的C—H键的羟基化、烯烃和芳烃的环氧化、含杂原子(N、S)化合物的氧化、乙醚裂解、N-脱烷基化、脱酰化和酚类的单电子氧化。UPO以H2O2为氧供体与电子受体,不需要任何辅因子,是目前最具发展潜力的氧化酶之一。然而,UPO的异源表达困难与选择性差的问题仍限制着UPO的发展。近两年,通过信号肽改造或更换的方法在UPO的异源表达方面取得了重要突破,对UPO结构功能关系的深入研究以及蛋白结构预测算法的发展也将助力UPO的分子改造,为解决UPO选择性差的问题奠定基础。本文聚焦于UPO的异源表达、选择性问题与H2O2原位再生,综述了UPO的最新发展以及存在的技术瓶颈,并对解决这些瓶颈问题的方案做出展望。
The selective insertion of oxygen species through unactivated C—H bonds is one of the most challenging tasks in organic synthesis.Fungal unspecific peroxygenases(UPOs)are a class of highly glycosylated thioheme enzymes that catalyze reactions including hydroxylation of unactivated C—H bonds in n-alkanes,epoxidation of alkenes and aromatics,oxidation of heteroatom(N,S)compounds,ether cleavage,N-dealkylation,deacylation and oneelectron oxidation of phenols.As one of the most promising oxidases in synthetic chemistry,UPOs use H_(2)O_(2)as the oxygen donor and the electron acceptor,and do not require cofactors other than heme.This paper reviews the classification and development process of UPOs,and focuses on the heterologous expression,selectivity engineering and H_(2)O_(2)in situ regeneration of UPOs.Since the first discovery of UPOs from Agrocybe aegerita in 2004,UPOs have attracted much attention due to the advantages described above.However,the difficulties of heterologous expression and poor selectivity of UPOs still limit their development.The difficulty of heterologous expression makes it hard to mine new variants of UPOs,and native UPOs are difficult to be characterized and applied in biocatalysis due to the slow growth rate of their hosts.In the past two years,important breakthroughs have been made in the heterologous expression of UPOs through the modification or replacement of signal peptides,revealing the important role of signal peptides in this process.However,the specific role of signal peptides in the secretory expression and three-dimensional structure formation of UPOs remain elusive.With the in-depth research on the mechanism of signal peptides affecting the heterologous expression of UPOs and the development of artificial intelligence(AI)algorithms,the combination of genome mining and signal peptide prediction will be the key for discovering new UPOs.The poor selectivity of UPOs also hinders the development and application of UPOs.This paper reviews different types of reactions that UPOs catalyze,and reveals the problem that UPOs have broad substrate range but poor selectivity.In-depth research on the structure-function relationship of UPOs and the development of protein structure prediction algorithms will help the engineering of UPOs and lay a foundation for solving the problem of poor substrate selectivity.This paper also compares several methods for the in situ regeneration of H_(2)O_(2),and concluded that the multi-enzyme cascade method is the most economical and practical method for the in situ regeneration of H_(2)O_(2).