NAD^(+)依赖型甲醇脱氢酶的改造和应用  

Engineering and application of NAD^(+)-dependent methanol dehydrogenase

作　　者：杨金星 王钰 YANG Jinxing;WANG Yu(Key Laboratory of Engineering Biology for Low-Carbon Manufacturing,Tianjin Institute of Industrial Biotechnology,Chinese Academy of Sciences,Tianjin 300308,China;National Technology Innovation Center of Synthetic Biology,Tianjin 300308,China)

机构地区：[1]中国科学院天津工业生物技术研究所低碳合成工程生物学重点实验室,天津300308 [2]国家合成生物技术创新中心,天津300308

出　　处：《生物加工过程》2024年第6期591-600,共10页Chinese Journal of Bioprocess Engineering

基　　金：中国科学院关键核心技术攻坚先导专项(XDC0110201);国家自然科学基金(32222004、32070083);中国科学院青年创新促进会(2021177)。

摘　　要：一碳原料因其来源广泛且价格低廉,已被视为极具前景的糖基碳源替代品,其中,甲醇更是凭借其价格低廉和便于运输的优势,在生物制造领域广受关注。甲醇脱氢酶(methanol dehydrogenase,Mdh)是甲醇生物利用的关键限速酶,负责将甲醇氧化为甲醛。目前,来自不同物种的Mdh已被相继鉴定,为甲醇生物转化提供了多种可供选择的催化元件,其中,烟酰胺腺嘌呤二核苷酸(nicotinamide adenine dinucleotide,NAD^(+))依赖型Mdh仅由单个基因编码,便于异源表达和改造,反应无需O 2参与,且催化过程伴随着还原力NADH生成,可为生物合成提供驱动力,因此被广泛用于构建人工的甲醇生物转化系统。然而,这类Mdh的催化活性相对较低,限制了甲醇的利用效率。通过新酶挖掘和定向进化等手段获得高活性的Mdh成为一碳原料生物转化利用领域的研究热点之一。本文系统综述NAD^(+)依赖型Mdh的鉴定和进化的相关成果,并简要介绍其在甲醇生物转化中的应用。One-carbon(C1)feedstocks are abundant and promising substitutes for sugar-based carbon sources.Among them,methanol has attracted significant attention in bio-manufacturing due to its low cost and ease of transportation.Methanol dehydrogenase(Mdh)is a key enzyme in methanol utilization,responsible for the oxidation of methanol to formaldehyde.Recently,a variety of Mdh were identified from different species,offering a range of catalytic options for methanol bioconversion.In particular,nicotinamide adenine dinucleotide(NAD^(+))-Mdh,which is encoded by a single gene,can be conveniently prepared through heterologous expression and transformation.More importantly,the corresponding catalytic reaction does not require the participation of oxygen,and NADH is simultaneously produced during the process to supply the necessary driving force for biosynthesis.Consequently,NAD^(+)-dependent Mdh is widely utilized in constructing artificial methanol bioconversion systems.However,the relatively low catalytic activity of NAD^(+)-dependent Mdh limits the efficiency of methanol utilization.Nowadays,one major focus in this field is to discover highly active Mdh through new enzyme mining and directed evolution,promoting the related bioconversion and utilization of single-carbon feedstocks.In this review,we summarized the recent scientific research with regards to the identification and evolution of NAD^(+)-dependent Mdh,as well as its applications in methanol bioconversion.

关 键 词：甲醇脱氢酶 甲醇生物转化 定向进化 

分 类 号：Q814[生物学—生物工程]