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Characteristics of a Streptomyces coelicolor A3(2) Extracellular Protein Targeting Chitin and Chitosan

机译:链霉菌链球菌A3(2)靶向壳多糖和壳聚糖的细胞外蛋白的特征。

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摘要

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 α-chitin, β-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.
机译:链霉菌天蓝色A3(2)几丁质酶G基因的上游,是一个小基因(名为chb3),其推导产物与上述寡脂链霉菌的CHB1蛋白具有37%的相同氨基酸。将chb3基因及其上游区域克隆到多拷贝载体中,并转化到无质粒的链霉菌链霉菌TK21菌株中。 CHB3蛋白(14.9 kDa)在存在葡萄糖,N-乙酰氨基葡萄糖,酵母提取物和几丁质的条件下,在生长过程中由葡萄链霉菌TK21转化子分泌。使用阴离子交换,疏水作用色谱和凝胶过滤将蛋白质纯化至均质。与CHB1相反,CHB3的目标是pH值为6.0的α-甲壳质,β-甲壳质和壳聚糖,但作用相对较松散。描述了各种类型的几丁质结合蛋白的底物特异性差异的生态意义。

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