{"title":"重组热溶酶的工程、表达、纯化和生产。","authors":"Kuniyo Inouye, Masayuki Kusano, Yasuhiko Hashida, Masashi Minoda, Kiyoshi Yasukawa","doi":"10.1016/S1387-2656(07)13003-9","DOIUrl":null,"url":null,"abstract":"<p><p>Thermolysin [EC 3.4.24.27] is a thermostable neutral zinc metalloproteinase originally identified in the culture broth of Bacillus thermoproteolyticus Rokko. Since the discovery in 1962, the enzyme has been extensively studied regarding its structure and catalytic mechanism. Today, thermolysin is a representative of zinc metalloproteinase and an attractive target in protein engineering to understand the catalytic mechanism, thermostability, and halophilicity. Thermolysin is used in industry, especially for the enzymatic synthesis of N-carbobenzoxy L-Asp-L-Phe methyl ester (ZDFM), a precursor of an artificial sweetener, aspartame. Generation of genetically engineered thermolysin with higher activity in the synthesis of ZDFM has been highly desired. In accordance with the expansion of studies on thermolysin, various strategies for its expression and purification have been devised and successfully used. In this review, we aim to outline recombinant thermolysins associated with their engineering, expression, purification, and production.</p>","PeriodicalId":79566,"journal":{"name":"Biotechnology annual review","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2007-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S1387-2656(07)13003-9","citationCount":"37","resultStr":"{\"title\":\"Engineering, expression, purification, and production of recombinant thermolysin.\",\"authors\":\"Kuniyo Inouye, Masayuki Kusano, Yasuhiko Hashida, Masashi Minoda, Kiyoshi Yasukawa\",\"doi\":\"10.1016/S1387-2656(07)13003-9\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Thermolysin [EC 3.4.24.27] is a thermostable neutral zinc metalloproteinase originally identified in the culture broth of Bacillus thermoproteolyticus Rokko. Since the discovery in 1962, the enzyme has been extensively studied regarding its structure and catalytic mechanism. Today, thermolysin is a representative of zinc metalloproteinase and an attractive target in protein engineering to understand the catalytic mechanism, thermostability, and halophilicity. Thermolysin is used in industry, especially for the enzymatic synthesis of N-carbobenzoxy L-Asp-L-Phe methyl ester (ZDFM), a precursor of an artificial sweetener, aspartame. Generation of genetically engineered thermolysin with higher activity in the synthesis of ZDFM has been highly desired. In accordance with the expansion of studies on thermolysin, various strategies for its expression and purification have been devised and successfully used. In this review, we aim to outline recombinant thermolysins associated with their engineering, expression, purification, and production.</p>\",\"PeriodicalId\":79566,\"journal\":{\"name\":\"Biotechnology annual review\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2007-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/S1387-2656(07)13003-9\",\"citationCount\":\"37\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biotechnology annual review\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1016/S1387-2656(07)13003-9\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biotechnology annual review","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1016/S1387-2656(07)13003-9","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 37
摘要
热溶酶[EC 3.4.24.27]是一种耐热的中性锌金属蛋白酶,最初在热溶蛋白芽孢杆菌培养液中发现。自1962年发现该酶以来,人们对其结构和催化机理进行了广泛的研究。目前,热溶酶是锌金属蛋白酶的代表,是蛋白质工程中研究催化机理、热稳定性和嗜盐性的重要靶点。热溶酶用于工业,特别是用于酶促合成n -碳苯氧基l - asp - l -苯丙氨酸甲酯(ZDFM),这是一种人工甜味剂阿斯巴甜的前体。在合成ZDFM的过程中,产生具有更高活性的基因工程热溶酶已被高度期望。随着对热溶素研究的不断深入,人们设计了多种热溶素的表达和纯化方法,并取得了成功。在这篇综述中,我们旨在概述重组热溶酶的工程、表达、纯化和生产。
Engineering, expression, purification, and production of recombinant thermolysin.
Thermolysin [EC 3.4.24.27] is a thermostable neutral zinc metalloproteinase originally identified in the culture broth of Bacillus thermoproteolyticus Rokko. Since the discovery in 1962, the enzyme has been extensively studied regarding its structure and catalytic mechanism. Today, thermolysin is a representative of zinc metalloproteinase and an attractive target in protein engineering to understand the catalytic mechanism, thermostability, and halophilicity. Thermolysin is used in industry, especially for the enzymatic synthesis of N-carbobenzoxy L-Asp-L-Phe methyl ester (ZDFM), a precursor of an artificial sweetener, aspartame. Generation of genetically engineered thermolysin with higher activity in the synthesis of ZDFM has been highly desired. In accordance with the expansion of studies on thermolysin, various strategies for its expression and purification have been devised and successfully used. In this review, we aim to outline recombinant thermolysins associated with their engineering, expression, purification, and production.