Purification, properties and cDNA cloning of glutamate decarboxylase in germinated faba bean (Vicia faba L.)

被引：22

作者：

Yang, Runqiang ^{[1
]}

Yin, Yongqi ^{[1
]}

Guo, Qianghui ^{[1
]}

Gu, Zhenxin ^{[1
]}

机构：

[1] Nanjing Agr Univ, Coll Food Sci & Technol, Nanjing 210095, Jiangsu, Peoples R China

来源：

FOOD CHEMISTRY | 2013年 / 138卷 / 2-3期

关键词：

Glutamate decarboxylase; Purification; Properties; cDNA cloning; Faba bean; GAMMA-AMINOBUTYRIC-ACID; DIAMINE OXIDASE; MOUSE-BRAIN; ACCUMULATION; GRAIN; EXPRESSION; CULTIVAR; ROOTS; GENE;

D O I：

10.1016/j.foodchem.2012.11.050

中图分类号：

O69 [应用化学];

学科分类号：

081704 ;

摘要：

Gamma-aminobutyric acid (GABA) is a non-protein amino acid with bioactive functions in humans. In this work, glutamate decarboxylase (EC 4.1.1.15, GAD) which is key in the GABA bioformation was purified from 5-day germinated faba beans and characterized. A single band was observed at 58 kDa using sodium dodecyl sulphate gel electrophoresis. GAD optimal activity was at pH 6.0 at 40 degrees C with a Km value for glutamic acid (Glu) of 2.63 mM. The enzyme was inhibited significantly by Cu2+, Fe3+, Mg2+, Ba2+, aminoxyacetate, EGTA, Na(2)EDTA, L-cysteine and beta-mercaptoethanol; and activated at low Ca2+ 0.2 mM. Using RT-PCR, the GAD cDNA was sequenced which indicated 1787 bp long, containing a 1527 bp open reading frame (ORF) that encoded 509 amino-acid peptides with a calculated molecular weight of 57.74 kDa and a pl of 5.41 (GenBank accession number: JX444699). Crown Copyright (C) 2012 Published by Elsevier Ltd. All rights reserved.

引用

页码：1945 / 1951

页数：7

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