Glutamate Dehydrogenase Functions in Glutamic Acid Metabolism and Stress Resistance in Pyropia haitanensis

Pyropia haitanensis is an important laver species in China. Its quality traits are closely related to the content of glutamic acid. Glutamate dehydrogenase (GDH) is a crucial enzyme in the glutamic acid metabolism. In this study, two GDH genes from P. h...

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Autores principales:	Shuang Li, Zhanru Shao, Chang Lu, Jianting Yao, Yongdong Zhou, Delin Duan
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	heterologous expression protein purification enzyme activity assay site-directed mutagenesis abiotic stresses Organic chemistry QD241-441
Acceso en línea:	https://doaj.org/article/a749fa1dc9eb4373824a32d4e9725ebd
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spelling	oai:doaj.org-article:a749fa1dc9eb4373824a32d4e9725ebd2021-11-25T18:27:18ZGlutamate Dehydrogenase Functions in Glutamic Acid Metabolism and Stress Resistance in <i>Pyropia haitanensis</i>10.3390/molecules262267931420-3049https://doaj.org/article/a749fa1dc9eb4373824a32d4e9725ebd2021-11-01T00:00:00Zhttps://www.mdpi.com/1420-3049/26/22/6793https://doaj.org/toc/1420-3049<i>Pyropia haitanensis</i> is an important laver species in China. Its quality traits are closely related to the content of glutamic acid. Glutamate dehydrogenase (GDH) is a crucial enzyme in the glutamic acid metabolism. In this study, two <i>GDH</i> genes from <i>P. haitanensis</i>, <i>PhGDH</i>1 and <i>PhGDH</i>2, were cloned and successfully expressed in <i>Escherichia coli</i>. The in vitro enzyme activity assay demonstrated that the catalytic activity of PhGDHs is mainly in the direction of ammonium assimilation. The measured <i>K<sub>m</sub></i> values of PhGDH1 for NADH, (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub>, and α-oxoglutarate were 0.12, 4.99, and 0.16 mM, respectively, while the corresponding <i>K<sub>m</sub></i> values of PhGDH2 were 0.02, 3.98, and 0.104 mM, respectively. Site-directed mutagenesis results showed that Gly<sup>193</sup> and Thr<sup>361</sup> were important catalytic residues for PhGDH2. Moreover, expression levels of both PhGDHs were significantly increased under abiotic stresses. These results suggest that PhGDHs can convert α-oxoglutarate to glutamic acid, and enhance the flavor and stress resistance of <i>P. haitanensis</i>.Shuang LiZhanru ShaoChang LuJianting YaoYongdong ZhouDelin DuanMDPI AGarticleheterologous expressionprotein purificationenzyme activity assaysite-directed mutagenesisabiotic stressesOrganic chemistryQD241-441ENMolecules, Vol 26, Iss 6793, p 6793 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	heterologous expression protein purification enzyme activity assay site-directed mutagenesis abiotic stresses Organic chemistry QD241-441
spellingShingle	heterologous expression protein purification enzyme activity assay site-directed mutagenesis abiotic stresses Organic chemistry QD241-441 Shuang Li Zhanru Shao Chang Lu Jianting Yao Yongdong Zhou Delin Duan Glutamate Dehydrogenase Functions in Glutamic Acid Metabolism and Stress Resistance in <i>Pyropia haitanensis</i>
description	<i>Pyropia haitanensis</i> is an important laver species in China. Its quality traits are closely related to the content of glutamic acid. Glutamate dehydrogenase (GDH) is a crucial enzyme in the glutamic acid metabolism. In this study, two <i>GDH</i> genes from <i>P. haitanensis</i>, <i>PhGDH</i>1 and <i>PhGDH</i>2, were cloned and successfully expressed in <i>Escherichia coli</i>. The in vitro enzyme activity assay demonstrated that the catalytic activity of PhGDHs is mainly in the direction of ammonium assimilation. The measured <i>K<sub>m</sub></i> values of PhGDH1 for NADH, (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub>, and α-oxoglutarate were 0.12, 4.99, and 0.16 mM, respectively, while the corresponding <i>K<sub>m</sub></i> values of PhGDH2 were 0.02, 3.98, and 0.104 mM, respectively. Site-directed mutagenesis results showed that Gly<sup>193</sup> and Thr<sup>361</sup> were important catalytic residues for PhGDH2. Moreover, expression levels of both PhGDHs were significantly increased under abiotic stresses. These results suggest that PhGDHs can convert α-oxoglutarate to glutamic acid, and enhance the flavor and stress resistance of <i>P. haitanensis</i>.
format	article
author	Shuang Li Zhanru Shao Chang Lu Jianting Yao Yongdong Zhou Delin Duan
author_facet	Shuang Li Zhanru Shao Chang Lu Jianting Yao Yongdong Zhou Delin Duan
author_sort	Shuang Li
title	Glutamate Dehydrogenase Functions in Glutamic Acid Metabolism and Stress Resistance in <i>Pyropia haitanensis</i>
title_short	Glutamate Dehydrogenase Functions in Glutamic Acid Metabolism and Stress Resistance in <i>Pyropia haitanensis</i>
title_full	Glutamate Dehydrogenase Functions in Glutamic Acid Metabolism and Stress Resistance in <i>Pyropia haitanensis</i>
title_fullStr	Glutamate Dehydrogenase Functions in Glutamic Acid Metabolism and Stress Resistance in <i>Pyropia haitanensis</i>
title_full_unstemmed	Glutamate Dehydrogenase Functions in Glutamic Acid Metabolism and Stress Resistance in <i>Pyropia haitanensis</i>
title_sort	glutamate dehydrogenase functions in glutamic acid metabolism and stress resistance in <i>pyropia haitanensis</i>
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/a749fa1dc9eb4373824a32d4e9725ebd
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Glutamate Dehydrogenase Functions in Glutamic Acid Metabolism and Stress Resistance in <i>Pyropia haitanensis</i>

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