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来自糖苷水解酶家族12的土曲霉内切-β-1,4-葡聚糖酶的表达、纯化、结晶及初步X射线衍射分析

Expression, purification, crystallization and preliminary X-ray diffraction analysis of Aspergillus terreus endo-β-1,4-glucanase from glycoside hydrolase family 12.

作者信息

Segato Fernando, Berto Gabriela L, Ares de Araújo Evandro, Muniz João Renato, Polikarpov Igor

机构信息

Instituto de Física de São Carlos, Universidade de São Paulo, Avenida Trabalhador Saocarlense 400, 13566-590 São Carlos-SP, Brazil.

出版信息

Acta Crystallogr F Struct Biol Commun. 2014 Feb;70(Pt 2):267-70. doi: 10.1107/S2053230X13034936. Epub 2014 Jan 28.

Abstract

Endoglucanases are important enzymes that are involved in the modification and degradation of cellulose. Filamentous fungi such as Aspergillus terreus are effective biomass degraders in nature owing to their capacity to produce an enzymatic arsenal of glycoside hydrolases, including endoglucanase from glycoside hydrolase family 12 (GH12). The A. terreus GH12 endoglucanase was cloned and overexpressed in A. nidulans, purified and crystallized. A single crystal was obtained from a solution consisting of 2 M ammonium sulfate, 5%(v/v) 2-propanol. X-ray diffraction data were collected to a resolution of 1.85 Å using synchrotron radiation and a preliminary molecular-replacement solution was obtained in the trigonal space group P3(2)21. The unit-cell parameters were a = b = 103.24, c = 48.96 Å.

摘要

内切葡聚糖酶是参与纤维素修饰和降解的重要酶。像土曲霉这样的丝状真菌在自然界中是有效的生物质降解者,因为它们能够产生包括来自糖苷水解酶家族12(GH12)的内切葡聚糖酶在内的糖苷水解酶酶库。土曲霉GH12内切葡聚糖酶在构巢曲霉中被克隆、过表达、纯化并结晶。从由2 M硫酸铵、5%(v/v)2-丙醇组成的溶液中获得了单晶。使用同步辐射收集了分辨率为1.85 Å的X射线衍射数据,并在三方空间群P3(2)21中获得了初步的分子置换解。晶胞参数为a = b = 103.24,c = 48.96 Å。

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