Efficient expression and characterization of a cold-active endo-1, 4-beta-glucanase from Citrobacter farmeri by co-expression of Myxococcus xanthus protein

文献类型： 外文期刊

第一作者： Bai, Xi

作者： Bai, Xi;Yuan, Xianjun;Li, Junfeng;Shao, Tao;Bai, Xi;Wen, Aiyou;Bai, Yunfeng

作者机构：

关键词： Cellulose degradation;Cellulose;Cold-active enzyme;Endoglucanases;Enzymatic properties;Escherichia coli;Expression;Novel expression vector;N-terminal fusion;Protein S-tag;Recombinant protein

期刊名称：ELECTRONIC JOURNAL OF BIOTECHNOLOGY （ 影响因子：2.8； 五年影响因子：3.379 ）

ISSN： 0717-3458

年卷期： 2016 年 24 卷

页码：

收录情况： SCI

摘要： Background: Cold-active endo-1, 4-beta-glucanase (EglC) can decrease energy costs and prevent product denaturation in biotechnological processes. However, the nature EglC from C. farmeri A1 showed very low activity (800 U/L). In an attempt to increase its expression level, C. farmeri EglC was expressed in Escherichia coli as an N-terminal fusion to protein S (ProS) from Myxococcus xanthus. Results: A novel expression vector, pET(ProS-EglC), was successfully constructed for the expression of C. farmeri EglC in E. coli. SDS-PAGE showed that the recombinant protein (ProS-EglC) was approximately 60 kDa. The activity of ProS-EglC was 12,400 U/L, which was considerably higher than that of the nature EglC (800 U/L). ProS-EglC was active at pH 6.5-pH 8.0, with optimum activity at pH 7.0. The recombinant protein was stable at pH 3.5-pH 6.5 for 30 min. The optimal temperature for activity of ProS-EglC was 30 degrees C-40 degrees C. It showed greater than 50% of maximum activity even at 5 degrees C, indicating that the ProS-EglC is a cold-active enzyme. Its activity was increased by Co2+ and Fe2+, but decreased by Cd2+, Zn2+, Li+, methanol, Triton-X-100, acetonitrile, Tween 80, and SDS. Conclusions: The ProS-EglC is promising in application of various biotechnological processes because of its cold-active characterizations. This study also suggests a useful strategy for the expression of foreign proteins in E. coli using a ProS tag. (C) 2016 Pontificia Universidad Catolica de Valparaiso. Production and hosting by Elsevier B.V. All rights reserved.

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