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微生物蛋白酶的分子与生物技术方面

Molecular and biotechnological aspects of microbial proteases.

作者信息

Rao M B, Tanksale A M, Ghatge M S, Deshpande V V

机构信息

Division of Biochemical Sciences, National Chemical Laboratory, Pune 411008, India.

出版信息

Microbiol Mol Biol Rev. 1998 Sep;62(3):597-635. doi: 10.1128/MMBR.62.3.597-635.1998.

Abstract

Proteases represent the class of enzymes which occupy a pivotal position with respect to their physiological roles as well as their commercial applications. They perform both degradative and synthetic functions. Since they are physiologically necessary for living organisms, proteases occur ubiquitously in a wide diversity of sources such as plants, animals, and microorganisms. Microbes are an attractive source of proteases owing to the limited space required for their cultivation and their ready susceptibility to genetic manipulation. Proteases are divided into exo- and endopeptidases based on their action at or away from the termini, respectively. They are also classified as serine proteases, aspartic proteases, cysteine proteases, and metalloproteases depending on the nature of the functional group at the active site. Proteases play a critical role in many physiological and pathophysiological processes. Based on their classification, four different types of catalytic mechanisms are operative. Proteases find extensive applications in the food and dairy industries. Alkaline proteases hold a great potential for application in the detergent and leather industries due to the increasing trend to develop environmentally friendly technologies. There is a renaissance of interest in using proteolytic enzymes as targets for developing therapeutic agents. Protease genes from several bacteria, fungi, and viruses have been cloned and sequenced with the prime aims of (i) overproduction of the enzyme by gene amplification, (ii) delineation of the role of the enzyme in pathogenecity, and (iii) alteration in enzyme properties to suit its commercial application. Protein engineering techniques have been exploited to obtain proteases which show unique specificity and/or enhanced stability at high temperature or pH or in the presence of detergents and to understand the structure-function relationships of the enzyme. Protein sequences of acidic, alkaline, and neutral proteases from diverse origins have been analyzed with the aim of studying their evolutionary relationships. Despite the extensive research on several aspects of proteases, there is a paucity of knowledge about the roles that govern the diverse specificity of these enzymes. Deciphering these secrets would enable us to exploit proteases for their applications in biotechnology.

摘要

蛋白酶是一类在生理作用和商业应用方面都占据关键地位的酶。它们兼具降解和合成功能。由于蛋白酶对生物体来说是生理必需的,所以在植物、动物和微生物等各种各样的来源中普遍存在。微生物是蛋白酶的一个有吸引力的来源,因为其培养所需空间有限,且易于进行基因操作。蛋白酶根据其作用于肽链末端还是远离末端,分别分为外切蛋白酶和内切蛋白酶。它们还根据活性位点官能团的性质分为丝氨酸蛋白酶、天冬氨酸蛋白酶、半胱氨酸蛋白酶和金属蛋白酶。蛋白酶在许多生理和病理生理过程中都起着关键作用。基于其分类,有四种不同类型的催化机制在起作用。蛋白酶在食品和乳制品行业有广泛应用。由于开发环境友好技术的趋势不断增加,碱性蛋白酶在洗涤剂和皮革行业有很大的应用潜力。将蛋白水解酶用作开发治疗剂的靶点正重新引起人们的兴趣。来自几种细菌、真菌和病毒的蛋白酶基因已被克隆和测序,主要目的是:(i)通过基因扩增过量生产该酶;(ii)阐明该酶在致病性中的作用;(iii)改变酶的性质以适应其商业应用。人们已利用蛋白质工程技术来获得在高温或极端pH值下,或在洗涤剂存在的情况下表现出独特特异性和/或更高稳定性的蛋白酶,并了解该酶的结构 - 功能关系。为了研究其进化关系,已对来自不同来源的酸性蛋白酶、碱性蛋白酶和中性蛋白酶的蛋白质序列进行了分析。尽管对蛋白酶的多个方面进行了广泛研究,但对于决定这些酶多种特异性的作用仍知之甚少。解开这些秘密将使我们能够利用蛋白酶在生物技术中的应用。

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