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| Cloning, Expression and Function Analysis of Methylenetetrahydrofolate Dehydrogenase from Mortierella alpina |
| WANG Hong chao, ZHANG Chen, CHEN Dian ning, QIAO Ju yuan, CHEN Hai qin, GU Zhen nan, ZHANG Hao, CHEN Wei, CHEN Yong quan |
| School of Food Science and Technology, Jiangnan University, Wuxi 214122, China |
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Abstract In folate metabolism, 5,10-methylenetetrahydrofolate is oxidized to 5,10-methenyltetrahydrofolate by the methylenetetrahydrofolate dehydrogenase (MTHFD) with the production of NADH or NADPH. To clarify the alternative NADPH sources in fatty acid synthesis, the heterologous expression vector for MTHFD was constructed under the skeleton of plasmid pET28a(+) and expressed in BL21 gold strain of E. coli. The enzymatic activity was investigated by VIS-UV spectroscopy. The M. alpina MTHFD can both catalyze NAD+ and NADP+, and is preferred to convert NADP+ to NADPH. The transcript level of MTHFD is up-regulated by nitrogen exhaustion, when M. alpina starts to accumulate lipids. This indicate the relationship between MTHFD and lipid metabolism is thus of major importance, and MTHFD may be an alternative NADPH source in fatty acid synthesis. The range of target candidates for genetic manipulation in M. alpina for obtaining strains with increased amounts of lipids were extended.
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Received: 16 May 2016
Published: 25 November 2016
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Cite this article:
WANG Hong chao, ZHANG Chen, CHEN Dian ning, QIAO Ju yuan, CHEN Hai qin, GU Zhen nan, ZHANG Hao, CHEN Wei, CHEN Yong quan. Cloning, Expression and Function Analysis of Methylenetetrahydrofolate Dehydrogenase from Mortierella alpina. China Biotechnology, 2016, 36(11): 23-29.
URL:
https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20161104 OR https://manu60.magtech.com.cn/biotech/Y2016/V36/I11/23
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