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从土壤宏基因组 DNA 中筛选得到一种新型耐有机溶剂酯酶的过表达及其性质研究

Overexpression and characterization of a new organic solvent-tolerant esterase derived from soil metagenomic DNA.

机构信息

Center for Biomedicine and Health, College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou 310015, China.

出版信息

Bioresour Technol. 2012 Jul;116:234-40. doi: 10.1016/j.biortech.2011.10.087. Epub 2011 Oct 31.

Abstract

In this study, an esterase, designated EstC23, was isolated from a soil metagenomic library. The protein was amenable to overexpression in Escherichia coli under control of the T7 promoter, resulting in expression of the active, soluble protein that constituted 30% of the total cell protein content. This enzyme showed optimal activity at 40 °C and retained about 50% maximal activity at 5-10 °C. EstC23 showed remarkable stability in up to 50% (v/v) benzene and alkanes (high logP solvents). When incubated for 7 days in the presence of 50% benzene or alkanes, the enzyme maintained its 2-3 fold elevated activity. The purified enzyme also cleaved sterically hindered esters of tertiary alcohols. These results indicate that EstC23 has potential for use in industrial processes.

摘要

在这项研究中,从土壤宏基因组文库中分离到一种酯酶,命名为 EstC23。该蛋白在 T7 启动子的控制下可在大肠杆菌中过表达,导致活性、可溶性蛋白的表达,占总细胞蛋白含量的 30%。该酶在 40°C 时表现出最佳活性,在 5-10°C 时保留约 50%的最大活性。EstC23 在高达 50%(v/v)的苯和烷烃(高 logP 溶剂)中表现出显著的稳定性。当在 50%的苯或烷烃存在下孵育 7 天时,酶保持其 2-3 倍的升高活性。纯化的酶还能切割位阻较大的叔醇酯。这些结果表明 EstC23 在工业过程中有潜在的用途。

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