title: Potential Application of the Oryza sativa Monodehydroascorbate Reductase Gene (OsMDHAR) to Improve the Stress Tolerance and Fermentative Capacity of Saccharomyces cerevisiae.
abstract: Monodehydroascorbate reductase (MDHAR; EC 1.6.5.4) is an important enzyme for ascorbate recycling. To examine whether heterologous expression of MDHAR from Oryza sativa (OsMDHAR) can prevent the deleterious effects of unfavorable growth conditions, we constructed a transgenic yeast strain harboring a recombinant plasmid carrying OsMDHAR (p426GPD::OsMDHAR). OsMDHAR-expressing yeast cells displayed enhanced tolerance to hydrogen peroxide by maintaining redox homoeostasis, proteostasis, and the ascorbate (AsA)-like pool following the accumulation of antioxidant enzymes and molecules, metabolic enzymes, and molecular chaperones and their cofactors, compared to wild-type (WT) cells carrying vector alone. The addition of exogenous AsA or its analogue isoascorbic acid increased the viability of WT and ara2Δ cells under oxidative stress. Furthermore, the survival of OsMDHAR-expressing cells was greater than that of WT cells when cells at mid-log growth phase were exposed to high concentrations of ethanol. High OsMDHAR expression also improved the fermentative capacity of the yeast during glucose-based batch fermentation at a standard cultivation temperature (30°C). The alcohol yield of OsMDHAR-expressing transgenic yeast during fermentation was approximately 25% (0.18 g·g-1) higher than that of WT yeast. Accordingly, OsMDHAR-expressing transgenic yeast showed prolonged survival during the environmental stresses produced during fermentation. These results suggest that heterologous OsMDHAR expression increases tolerance to reactive oxygen species-induced oxidative stress by improving cellular redox homeostasis and improves survival during fermentation, which enhances fermentative capacity.
authors.affiliation: School of Life Sciences, BK21 Plus KNU Creative BioResearch Group, Kyungpook National University, Daegu, Republic of Korea.
authors.forename: Il-Sup
authors.initials: IS
authors.lastname: Kim
authors.affiliation: Division of Biological Sciences, University of California San Diego, La Jolla, California, United States of America.
authors.forename: Young-Saeng
authors.initials: YS
authors.lastname: Kim
authors.affiliation: Highland Agriculture Research Institute, National Institute of Crop Science, Rural Development Administration, Pyeongchang, Republic of Korea.
authors.forename: Yul-Ho
authors.initials: YH
authors.lastname: Kim
authors.affiliation: Division of Polar Life Sciences, Korea Polar Research Institute, Incheon, Republic of Korea.
authors.forename: Ae-Kyung
authors.initials: AK
authors.lastname: Park
authors.affiliation: Division of Polar Life Sciences, Korea Polar Research Institute, Incheon, Republic of Korea.
authors.forename: Han-Woo
authors.initials: HW
authors.lastname: Kim
authors.affiliation: Division of Polar Life Sciences, Korea Polar Research Institute, Incheon, Republic of Korea.
authors.forename: Jun-Hyuk
authors.initials: JH
authors.lastname: Lee
authors.affiliation: School of Life Sciences, BK21 Plus KNU Creative BioResearch Group, Kyungpook National University, Daegu, Republic of Korea.
authors.forename: Ho-Sung
authors.initials: HS
authors.lastname: Yoon
mesh_terms: D000431:Ethanol / Q000378:metabolism; D015870:Gene Expression; D009247:NADH, NADPH Oxidoreductases / Q000096:biosynthesis / Q000235:genetics; D030781:Organisms, Genetically Modified / Q000235:genetics / Q000378:metabolism; D012275:Oryza / Q000201:enzymology / Q000235:genetics*; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D013312:Stress, Physiological
chemical_list: D010940:Plant Proteins; D000431:Ethanol; D009247:NADH, NADPH Oxidoreductases; C023927:monodehydroascorbate reductase (NADH)