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| Cloning and Enzyme Kinetics Analysis of FgGDH Gene from Fusarium graminearum |
| LUO Qiong*, HE Yong*, YAN Lu, ZENG Hui, XIA Qing-Shan, WANG Yan, LIN Jian-Zhong**, LIU Xuan-Ming** |
| College of Biology/Hunan Province Key Laboratory of Plant Functional Genomics and Developmental Regulation, Hunan University, Changsha 410082, China |
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Abstract Glutamate dehydrogenase (GDH) is an enzyme widely found in organisms related to nitrogen metabolism, and GDH genes of lower organisms are often used for the improvement of nitrogen assimilation efficiency in crops. In order to explore the high quality GDH genes for genetic improvement of crops, FgGDH gene was cloned from Fusarium graminearum, the protein FgGDH was prokaryotic expressed and purified, and its enzymatic kinetics characteristics were determined in vitro in this study. The results showed that the aminating activity of FgGDH was significantly higher than that of deaminating activity, indicating that FgGDH is more inclined to catalyze NH4+ and α-ketoglutaric acid (2-OG) to generate glutamic acid and promotes the assimilation of NH4+. In addition, the Km values of FgGDH for NH4+ and 2-OG were (3.71±0.21) mmol/L and (6.44±0.32) mmol/L, respectively, which were significantly lower than those of OsGDH4 in rice. These results indicate that the affinity of FgGDH for NH4+ and 2-OG is marked higher than those of endogenous OsGDH4 in rice, and FgGDH has a higher ammonia assimilation efficiency. This study provides a theoretical basis for the genetic improvement of nitrogen assimilation efficiency in rice and other crops by using FgGDH gene.
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Received: 18 November 2020
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Corresponding Authors:
** jianzhlin@hnu.edu.cn;xml05@hnu.edu.cn
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| About author:: * These authors contributed equally to this work |
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