The Role of Glyceraldehyde-3-Phosphate Dehydrogenases in NADPH Supply in the Oleaginous Filamentous Fungus Mortierella alpina

Shunxian Wang; Shunxian Wang; Haiqin Chen; Haiqin Chen; Xin Tang; Xin Tang; Hao Zhang; Hao Zhang; Hao Zhang; Hao Zhang; Guangfei Hao; Guangfei Hao; Wei Chen; Wei Chen; Wei Chen; Wei Chen; Yong Q. Chen; Yong Q. Chen; Yong Q. Chen; Yong Q. Chen; Yong Q. Chen

doi:10.3389/fmicb.2020.00818

Frontiers in Microbiology (Apr 2020)

The Role of Glyceraldehyde-3-Phosphate Dehydrogenases in NADPH Supply in the Oleaginous Filamentous Fungus Mortierella alpina

Shunxian Wang,
Shunxian Wang,
Haiqin Chen,
Haiqin Chen,
Xin Tang,
Xin Tang,
Hao Zhang,
Hao Zhang,
Hao Zhang,
Hao Zhang,
Guangfei Hao,
Guangfei Hao,
Wei Chen,
Wei Chen,
Wei Chen,
Wei Chen,
Yong Q. Chen,
Yong Q. Chen,
Yong Q. Chen,
Yong Q. Chen,
Yong Q. Chen

Affiliations

Shunxian Wang: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, China
Shunxian Wang: School of Food Science and Technology, Jiangnan University, Wuxi, China
Haiqin Chen: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, China
Haiqin Chen: School of Food Science and Technology, Jiangnan University, Wuxi, China
Xin Tang: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, China
Xin Tang: School of Food Science and Technology, Jiangnan University, Wuxi, China
Hao Zhang: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, China
Hao Zhang: School of Food Science and Technology, Jiangnan University, Wuxi, China
Hao Zhang: National Engineering Research Center for Functional Food, Jiangnan University, Wuxi, China
Hao Zhang: Wuxi Translational Medicine Research Center and Jiangsu Translational Medicine Research Institute Wuxi Branch, Wuxi, China
Guangfei Hao: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, China
Guangfei Hao: School of Food Science and Technology, Jiangnan University, Wuxi, China
Wei Chen: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, China
Wei Chen: School of Food Science and Technology, Jiangnan University, Wuxi, China
Wei Chen: National Engineering Research Center for Functional Food, Jiangnan University, Wuxi, China
Wei Chen: Beijing Innovation Centre of Food Nutrition and Human Health, Beijing Technology and Business University, Beijing, China
Yong Q. Chen: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, China
Yong Q. Chen: School of Food Science and Technology, Jiangnan University, Wuxi, China
Yong Q. Chen: National Engineering Research Center for Functional Food, Jiangnan University, Wuxi, China
Yong Q. Chen: Wuxi Translational Medicine Research Center and Jiangsu Translational Medicine Research Institute Wuxi Branch, Wuxi, China
Yong Q. Chen: Department of Cancer Biology, Wake Forest School of Medicine, Winston-Salem, NC, United States

DOI: https://doi.org/10.3389/fmicb.2020.00818
Journal volume & issue: Vol. 11

Abstract

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Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a highly conserved enzyme within the glycolytic pathway. GAPDH catalyzes the transformation of glyceraldehyde 3-phosphate to glycerate-1, 3-biphosphate, a process accompanied by the production of NADH. Its role in the NADPH production system of the oleaginous filamentous fungus Mortierella alpina was explored. Two copies of genes encoding GAPDH were characterized, then endogenously overexpressed and silenced through Agrobacterium tumefaciens-mediated transformation methods. The results showed that the lipid content of the overexpression strain, MA-GAPDH1, increased by around 13%. RNA interference of GAPDH1 and GAPDH2 (MA-RGAPDH1 and MA-RGAPDH2) greatly reduced the biomass of the fungus. The lipid content of MA-RGAPDH2 was found to be about 23% higher than that of the control. Both of the lipid-increasing transformants showed a higher NADPH/NADP ratio. Analysis of metabolite and enzyme expression levels revealed that the increased lipid content of MA-GAPDH1 was due to enhanced flux of glyceraldehyde-3-phosphate to glycerate-1, 3-biphosphate. MA-RGAPDH2 was found to strengthen the metabolic flux of dihydroxyacetone phosphate to glycerol-3-phosphate. Thus, GAPDH1 contributes to NADPH supply and lipid accumulation in M. alpina, and has a distinct role from GAPDH2.

Published in Frontiers in Microbiology

ISSN: 1664-302X (Online)
Publisher: Frontiers Media S.A.
Country of publisher: Switzerland
LCC subjects: Science: Microbiology
Website: http://www.frontiersin.org/journals/microbiology

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