| Orginal Article |
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| Effects of MTHFD1 Overexpression on Lipid Synthesis in the Oleaginous Fungus, Mortierella alpina |
Qi-zai WANG,Hong-chao WANG( ),Hai-qin CHEN,Jian-xin ZHAO,Hao ZHANG,Wei CHEN,Yong-quan CHEN |
| School of Food Science and Technology, Jiangnan University, Wuxi 214122, China |
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Abstract Mortierella alpina is a filamentous fungus with a high polyunsaturated fatty acids content,but its sources of NADPH needed for lipid synthesis are not been studied completely. The binary vector pBIG2-ura5s-MTHFD1 was constructed for the transformation of the methylenetetrahydrofolate dehydrogenase(MTHFD1)into the M.alpina (uracil auxotrophic) strain by Agrobacterium tumefaciens-mediated transformation (ATMT). PCR analysis identified the presence of the MTHFD1 overexpression cassette in the genome, and qPCR analysis showed that the transcript levels of the MTHFD1 gene in MTHFD1 overexpression strain (MA-MTHFD1) were significantly increased compared to controls. The results showed that MTHFD1 overexpression influenced lipid synthesis significantly. In comparsion to prototrophic M. alpina, the total fatty acid(TFA)level increased by about 40.13% and NADPH level simultaneously increased by 26.45% in MA-MTHFD1. Moreover, two key enzymes ME and IDH in NADPH synthesis were up-regulated. Based on the results, MTHFD1 plays an important role in NADPH generation during lipid synthesis in M. alpina. A foundation for uncovering mechanism of lipogenesis in M. alpina was established.
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Received: 05 March 2018
Published: 12 October 2018
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Corresponding Authors:
Hong-chao WANG
E-mail: hcwang@jiangnan.edu.cn
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