Expression, purification, and characterization of recombinant mangrove glutamine synthetase

文献类型： 外文期刊

第一作者： Zhao, Wei

作者： Zhao, Wei;Tian, Yongsheng;Fu, Xiaoyan;Zhu, Bo;Xue, Yong;Gao, Jianjie;Han, Hong-Juan;Peng, Rihe;Yao, Quan-Hong;Yang, Jun

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关键词： Glutamine synthetase (GS);Ammonium assimilation;Mangrove (Avicennia marina);PPT-sensitive

期刊名称：MOLECULAR BIOLOGY REPORTS （ 影响因子：2.316； 五年影响因子：2.357 ）

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收录情况： SCI

摘要： To expand our knowledge about the relationship of nitrogen use efficiency and glutamine synthetase (GS) activity in the mangrove plant, a cytosolic GS gene from Avicennia marina has been heterologously expressed in and purified from Escherichia coli. Synthesis of the mangrove GS enzyme in E. coli was demonstrated by functional genetic complementation of a GS deficient mutant. The subunit molecular mass of GSI was similar to 40 kDa. Optimal conditions for biosynthetic activity were found to be 35 degrees C at pH 7.5. The Mg2+-dependent biosynthetic activity was strongly inhibited by Ni2+, Zn2+, and Al3+, whereas was enhanced by Co2+. The apparent Km values of AmGLN1 for the substrates in the biosynthetic assay were 3.15 mM for glutamate, and 2.54 mM for ATP, 2.80 mM for NH4+ respectively. The low affinity kinetics of AmGLN1 apparently participates in glutamine synthesis under the ammonium excess conditions.

分类号： Q7

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