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从瘤胃宏基因组文库中分离α-葡萄糖醛酸酶酶。

Isolation of α-glucuronidase enzyme from a rumen metagenomic library.

机构信息

USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710, USA.

出版信息

Protein J. 2012 Mar;31(3):206-11. doi: 10.1007/s10930-012-9391-z.

Abstract

α-Glucuronidase enzymes play an essential role in the full enzymatic hydrolysis of hemicellulose. Up to this point, all genes encoding α-glucuronidase enzymes have been cloned from individual, pure culture strains. Using a high-throughput screening strategy, we have isolated the first α-glucuronidase gene (rum630-AG) from a mixed population of microorganisms. The gene was subcloned into a prokaryotic vector, and the enzyme was overexpressed and biochemically characterized. The RUM630-AG enzyme had optimum activity at pH 6.5 and 40 °C. When birchwood xylan was used as substrate, the RUM630-AG functioned synergistically with an endoxylanase enzyme to hydrolyze the substrate.

摘要

α-葡萄糖醛酸酶在半纤维素的完全酶解中起着至关重要的作用。到目前为止,所有编码α-葡萄糖醛酸酶的基因都是从单个纯培养菌株中克隆出来的。我们使用高通量筛选策略,从混合微生物群体中分离到第一个α-葡萄糖醛酸酶基因(rum630-AG)。该基因被亚克隆到原核载体中,过表达并进行了生化特性分析。RUM630-AG 酶在 pH6.5 和 40°C 时具有最佳活性。当使用桦木木聚糖作为底物时,RUM630-AG 与内切木聚糖酶协同作用水解底物。

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