Isolation and biochemical characterization of a metagenome-derived 3-deoxy-d-arabino-heptulosonate-7-phosphate synthase gene from subtropical marine mangrove wetland sediments

doi:10.1186/s13568-019-0742-4

	Isolation and biochemical characterization of a metagenome-derived 3-deoxy-d-arabino-heptulosonate-7-phosphate synthase gene from subtropical marine mangrove wetland sediments
	Zhao,Huaxian 1; Gao,Hua 1; Ji,Kai 1; Yan,Bing 2; Li,Quanwen 1; Mo,Shuming 1; Zheng,Minggang 4; Ou,Qian 1; Wu,Bo 1; Li,Nan 3
刊名	AMB Express
	2019-02-04
卷号	9 期号:1
关键词	3-Deoxy-d-arabino-heptulosonate-7-phosphate synthase Novel gene Metagenomic library Subtropical marine mangrove sediment Biochemical characterization
ISSN号	2191-0855
DOI	10.1186/s13568-019-0742-4
通讯作者	Li,Nan(nli@yic.ac.cn) ; Jiang,Chengjian(jiangcj0520@gmail.com)
英文摘要	Abstract3-Deoxy-d-arabino-heptulosonate-7-phosphate synthase (DAHPS) is a key rate-limiting enzyme in aromatic amino acid anabolism. A new Iβ-type DAHPS gene (aro1A) was identified in a metagenomic library from subtropical marine mangrove sediment. The gene encoded a polypeptide composed of 272 amino acids and had a maximum similarity of 52.4% to a known DAHPS at the amino acid level. Multiple sequence alignment, homologous modeling, and molecular docking showed that Aro1A had the typical (β/α)8 barrel-shaped catalytic structural domain of DAHPS. The motifs and amino acid residues involved in the combination of substrates and metal ligand were highly conservative with the known DAHPS. The putative DAHPS gene was subcloned into a pET-30a(+) vector and was overexpressed in Escherichia coli Rosetta (DE3) cells. The recombinant protein was purified to homogeneity. The maximum activity for the recombinant Aro1A protein occurred at pH 8.0 and 40?°C. Ba2+ and Ca2+ stimulated the activity of Aro1A protein. The enzyme showed high affinity and catalytic efficiency (KmPEP?=?19.58?μM, VmaxPEP? =?29.02?μM?min?1, and kcatPEP/KmPEP?=?0.88?s?1?μM?1) under optimal reaction conditions. The enzymatic property of Aro1A indicates its potential in aromatic amino acid industrial production.
语种	英语
WOS记录号	BMC:10.1186/S13568-019-0742-4
内容类型	期刊论文
源URL	[http://ir.yic.ac.cn/handle/133337/34490]
专题	中国科学院烟台海岸带研究所
通讯作者	Li,Nan; Jiang,Chengjian
作者单位	1. 2. 3. 4.
推荐引用方式 GB/T 7714	Zhao,Huaxian,Gao,Hua,Ji,Kai,et al. Isolation and biochemical characterization of a metagenome-derived 3-deoxy-d-arabino-heptulosonate-7-phosphate synthase gene from subtropical marine mangrove wetland sediments[J]. AMB Express,2019,9(1).
APA	Zhao,Huaxian.,Gao,Hua.,Ji,Kai.,Yan,Bing.,Li,Quanwen.,...&Jiang,Chengjian.(2019).Isolation and biochemical characterization of a metagenome-derived 3-deoxy-d-arabino-heptulosonate-7-phosphate synthase gene from subtropical marine mangrove wetland sediments.AMB Express,9(1).
MLA	Zhao,Huaxian,et al."Isolation and biochemical characterization of a metagenome-derived 3-deoxy-d-arabino-heptulosonate-7-phosphate synthase gene from subtropical marine mangrove wetland sediments".AMB Express 9.1(2019).