Cloning and identification of a novel NhaD-type Na+/H+ antiporter from metagenomic DNA of the halophilic bacteria in soil samples around Daban Salt Lake

文献类型： 外文期刊

第一作者： Zhang, Hua

作者： Zhang, Hua;Wang, Zhenhui;Mu, Ren;Zou, Zhi;Yuan, Kun;Wang, Yuekun;Wu, Haiping;Yang, Lifu;Zhang, Hua;Wang, Zhenhui;Mu, Ren;Zou, Zhi;Yuan, Kun;Wang, Yuekun;Wu, Haiping;Yang, Lifu;Zhang, Hua;Mu, Ren;Wu, Haiping;Yang, Lifu;Wang, Lei;Jiang, Juquan;Jiang, Juquan

作者机构：

关键词： Cloning;Identification;Novel NhaD-type Na+/H+ antiporter;Metagenomic DNA;Halophilic bacteria

期刊名称：EXTREMOPHILES （ 影响因子：2.395； 五年影响因子：2.668 ）

ISSN： 1431-0651

年卷期： 2014 年 18 卷 1 期

页码：

收录情况： SCI

摘要： In this study, metagenomic DNA was screened for the Na+/H+ antiporter gene from the halophilic bacteria in Daban Salt Lake by selection in Escherichia coli KNabc lacking three major Na+/H+ antiporters. One gene designated as Hb_nhaD encoding a novel NhaD-type Na+/H+ antiporter was finally cloned. The presence of Hb_NhaD conferred tolerance of E. coli KNabc to up to 0.5 M NaCl and 0.2 M LiCl, and an alkaline pH. Hb_NhaD has the highest identity (70.6 %) with a putative NhaD-type Na+/H+ antiporter from an uncharacterized Clostridiaceae species, and also has lower identity with known NhaD-type Na+/H+ antiporters from Halomonas elongata (20.8 %), Alkalimonas amylolytica (19.0 %), Vibrio parahaemolyticus (18.9 %) and Vibrio cholerae (18.7 %). pH-dependent Na+(Li+)/H+ antiport activity was detected from everted membrane vesicles prepared from E. coli KNabc carrying Hb_nhaD. Hb_NhaD exhibited very high Na+(Li+)/H+ antiport activity over a wide pH range from 6.5 to 9.0 with the highest activity at pH 7.0 which is significantly different from those of the above known NhaD-type Na+/H+ antiporters. Also, the apparent K (m) values of Hb_NhaD for Na+ and Li+ at pH 7.0 were determined to be 1.31 and 2.16, respectively. Based on the above results, we proposed that Hb_NhaD should be categorized as a novel NhaD-type Na+/H+ antiporter

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