天蓝色链霉菌A3(2)一种靶向几丁质和壳聚糖的细胞外蛋白的特性
Characteristics of a Streptomyces coelicolor A3(2) extracellular protein targeting chitin and chitosan.
作者信息
Saito A, Miyashita K, Biukovic G, Schrempf H
机构信息
FB Biologie/Chemie, Universität Osnabrück, 49069 Osnabrück, Germany.
出版信息
Appl Environ Microbiol. 2001 Mar;67(3):1268-73. doi: 10.1128/AEM.67.3.1268-1273.2001.
Abstract
Upstream of the Streptomyces coelicolor A3(2) chitinase G gene, a small gene (named chb3) is located whose deduced product shares 37% identical amino acids with the previously described CHB1 protein from Streptomyces olivaceoviridis. The chb3 gene and its upstream region were cloned in a multicopy vector and transformed into the plasmid-free Streptomyces lividans TK21 strain. The CHB3 protein (14.9 kDa) was secreted by the S. lividans TK21 transformant during growth in the presence of glucose, N-acetylglucosamine, yeast extract, and chitin. The protein was purified to homogeneity using anionic exchange, hydrophobic interaction chromatographies, and gel filtration. In contrast to CHB1, CHB3 targets alpha-chitin, beta-chitin, and chitosan at pH 6.0 but does so relatively loosely. The ecological implications of the divergence of substrate specificity of various types of chitin-binding proteins are described.
摘要
在天蓝色链霉菌A3(2)几丁质酶G基因的上游，定位了一个小基因（命名为chb3），其推导产物与先前描述的来自橄榄绿链霉菌的CHB1蛋白有37%的相同氨基酸。chb3基因及其上游区域被克隆到一个多拷贝载体中，并转化到无质粒的变铅青链霉菌TK21菌株中。在葡萄糖、N-乙酰葡糖胺、酵母提取物和几丁质存在的情况下生长时，CHB3蛋白（14.9 kDa）由变铅青链霉菌TK21转化体分泌。使用阴离子交换、疏水相互作用色谱法和凝胶过滤将该蛋白纯化至同质。与CHB1不同，CHB3在pH 6.0时靶向α-几丁质、β-几丁质和壳聚糖，但结合相对松散。描述了各种类型几丁质结合蛋白底物特异性差异的生态学意义。
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