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Abstract
We performed metabolic engineering on the budding yeast Saccharomyces cerevisiae for enhanced production of succinic acid. Aerobic succinic acid production in S. cerevisiae was achieved by disrupting the SDH1 and SDH2 genes, which encode the catalytic subunits of succinic acid dehydrogenase. Increased succinic acid production was achieved by eliminating the ethanol biosynthesis pathways. Metabolic profiling analysis revealed that succinic acid accumulated intracellularly following disruption of the SDH1 and SDH2 genes, which suggests that enhancing the export of intracellular succinic acid outside of cells increases succinic acid production in S. cerevisiae. The mae1 gene encoding the Schizosaccharomyces pombe malic acid transporter was introduced into S. cerevisiae, and as a result, succinic acid production was successfully improved. Metabolic profiling analysis is useful in producing chemicals for metabolic engineering of microorganisms.
Graphical Abstract
In this study, we examined succinic acid production by Saccharomyces cerevisiae and improved the productivity by metabolic engineering based on metabolic profiling analysis with CE-TOFMS.
Acknowledgments
The genome DNA clone for S. pombe FYG16_m23 and S. cerevisiae expression vector pNV11 was provided by the National Bio-Resource Project (NBRP) of the Ministry of Education, Culture, Sports, Science and Technology of Japan. We thank Ms Yuki Mori and Mr Tatsuya Itoga for CE-TOFMS analysis.
Funding
This work was supported financially by a Grant-in-Aid for Scientific Research (A) (No. 24246134, to S.H.) from the Japan Society for the Promotion of Science.