Expression and biochemical characterization of α-ketoglutarate decarboxylase from cyanobacterium Synechococcus sp PCC7002

被引：8

作者：

Lei, Guofeng ^{[1
]}

Wang, Xiaoqin ^{[1
]}

Lai, Chongde ^{[1
,2
]}

Li, Zhi-Min ^{[3
]}

Zhang, Wei ^{[1
]}

Xie, Congcong ^{[1
]}

Wang, Fei ^{[1
,2
]}

Wu, Xiaoyu ^{[1
,2
]}

Li, Zhimin ^{[1
,2
,4
]}

机构：

[1] Jiangxi Agr Univ, Coll Biosci & Bioengn, Jiangxi Engn Lab Dev & Utilizat Agr Microbial Res, Nanchang 330045, Jiangxi, Peoples R China

[2] Collaborat Innovat Ctr Postharvest Key Technol &, Nanchang 330045, Jiangxi, Peoples R China

[3] Jiangxi Agr Univ, Coll Sci, Nanchang 330045, Jiangxi, Peoples R China

[4] Jiangxi Agr Univ, Jiangxi Key Lab Conservat & Utilizat Fungal Resou, Nanchang 330045, Jiangxi, Peoples R China

来源：

INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES | 2018年 / 114卷

基金：

中国国家自然科学基金;

关键词：

alpha-Ketoglutarate decarboxylase; Synechococcus sp PCC7002; Protein expression; Kinetic characterization; Catalytic function; TRICARBOXYLIC-ACID CYCLE; DIPHOSPHATE-DEPENDENT ENZYME; BLUE-GREEN-ALGAE; SP PCC 6803; ESCHERICHIA-COLI; 2-OXOGLUTARATE DECARBOXYLASE; EUGLENA-GRACILIS; PHOTOSYNTHETIC PRODUCTION; SYNTHASE; IDENTIFICATION;

D O I：

10.1016/j.ijbiomac.2018.03.112

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

alpha-Ketoglutarate decarboxylase (alpha-KGD), one member of alpha-keto acid decarboxylases, catalyzing non-oxidative decarboxylation of alpha-ketoglutarate to form succinic semialdehyde, was proposed to play critical role in completing tricarboxylic acid (TCA) cycle of cyanobacteria. Although the catalytic function of alpha-KGD from Synechococcus sp. PCC7002 was demonstrated previously, there was no detailed biochemical characterization of alpha-KGD from Synechococcus sp. PCC7002 yet. In this study, the gene encoding alpha-KGD from Synechococcus sp. PCC7002 was amplified and soluble expression of recombinant alpha-KGD was achieved by coexpressing with pTf16 chaperone plasmid in E. coif BL21 (DE3). Kinetic analysis showed that the activity of alpha-KGD was dependent on cofactors of thiamine pyrophosphate and divalent cation. Meanwhile this alpha-KGD was specific for alpha-ketoglutarate with respect to the decarboxylation activity despite of the pretty low activity of acetolactate synthase. The catalytic efficiency of alpha-KGD (the values of k(cat) and k(cat)/K-m for alpha-ketoglutarate were 1.2 s(-1) and 6.3 x 10(3) M-1 s(-1), respectively) might provide evidence for its physiological role in TCA cycle of Synechococcus sp. PCC7002. (C) 2018 Elsevier B.V. All rights reserved.

引用

页码：188 / 193

页数：6

共 34 条

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