Abstract
Coenzyme Q (CoQ) is an essential component of the electron transport system that produces ATP in nearly all living cells. CoQ10 is a popular commercial food supplement around the world, and demand for efficient production of this molecule has increased in recent years. In this study, we explored CoQ10 production in the fission yeast Schizosaccharomyces pombe. We found that CoQ10 level was higher in stationary phase than in log phase, and that it increased when the cells were grown in a low concentration of glucose, in maltose, or in glycerol/ethanol medium. Because glucose signaling is mediated by cAMP, we evaluated the involvement of this pathway in CoQ biosynthesis. Loss of Pka1, the catalytic subunit of cAMP-dependent protein kinase, increased production of CoQ10, whereas loss of the regulatory subunit Cgs1 decreased production. Manipulation of other components of the cAMP-signaling pathway affected CoQ10 production in a consistent manner. We also found that glycerol metabolism was controlled by the cAMP/PKA pathway. CoQ10 production by the S. pombe ∆pka1 reached 0.98 mg/g dry cell weight in medium containing a non-fermentable carbon source [2% glycerol (w/v) and 1% ethanol (w/v) supplemented with 0.5% casamino acids (w/v)], twofold higher than the production in wild-type cells under normal growth conditions. These findings demonstrate that carbon source, growth phase, and the cAMP-signaling pathway are important factors in CoQ10 production in S. pombe.
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Acknowledgments
S. pombe kinase gene deletion mutants were kindly provided by Dr. M. Balasubramanian. Δpka1 (JZ633) was provided by Dr. M. Yamamoto. The authors also thank Dr. T. Nakagawa, H. Okazaki, and all other members of the laboratory for helpful discussions and support.
Funding
This work was partly supported by a grant-in-aid from the Ministry of Education, Culture, Sports, Science, and Technology of Japan [#25660059, #17H03806, and #24380056] to M.K., [#15K07360 and #18K05393] to T.K., [#18K14377] to I.N., [#18K05438] to Y.M., and by the Science and Technology Research Promotion Program for Agriculture, Forestry, Fisheries, and Food Industry [#957613] to M.K. The authors thank the faculty of Life and Environmental Sciences in Shimane University for help in financial supports for publishing this report.
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Nishida, I., Yokomi, K., Hosono, K. et al. CoQ10 production in Schizosaccharomyces pombe is increased by reduction of glucose levels or deletion of pka1. Appl Microbiol Biotechnol 103, 4899–4915 (2019). https://doi.org/10.1007/s00253-019-09843-7
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DOI: https://doi.org/10.1007/s00253-019-09843-7