Engineering, expression, purification, and production of recombinant thermolysin
Engineering, expression, purification, and production of recombinant thermolysin
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DOI：
10.1016/s1387-2656(07)13003-9
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发表时间：
2007-01-01
期刊：
影响因子：
--
通讯作者：
Yasukawa, Kiyoshi
中科院分区：
文献类型：
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作者：
Inouye, Kuniyo;Kusano, Masayuki;Yasukawa, Kiyoshi
嗜热菌蛋白酶[EC 3.4.24.27]是一种热稳定中性锌金属蛋白酶，最初在热蛋白水解芽孢杆菌Rokko的培养液中鉴定。自1962年发现以来，人们对其结构和催化机制进行了广泛的研究。嗜热菌蛋白酶是锌金属蛋白酶的代表，是蛋白质工程研究中的一个重要靶点。嗜热菌蛋白酶在工业中使用，特别是用于N-苄氧羰基L-Asp-L-Phe甲酯（ZDFM）的酶促合成，ZDFM是人工甜味剂甜菊糖的前体。在ZDFM的合成中，具有更高活性的基因工程嗜热菌蛋白酶的产生一直是高度期望的。随着嗜热菌蛋白酶研究的不断深入，人们设计并成功应用了多种表达和纯化策略。在这篇综述中，我们的目的是概述重组嗜热菌蛋白酶与工程，表达，纯化和生产。
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.