高产GSH酿酒酵母突变株Y518的生理代谢研究-微生物学专业论文.docxVIP

高产 GSH 酿酒酵母突变株 Y518 的生理代谢研究 英文摘要 II Analysis of physiological and matabolic changes in GSH-overaccumulating Saccharomyces cerevisiae mutant Y518 Abstract In this paper, physiological metabolism of GSH-overaccumulating saccharomyces cerevisiae mutant Y518 was studied from the aspects of physiological features and gene expression. The results indicated that the mutant had some defect in respiratory function and decreased energy metabolism. In addition, the mutant suffered oxidative stress caused by enhanced ROS, which was eliminated by increased GSH level and expressions of related gene. （1）The objective of this study was to analyze physiological changes of yeast mutant Y518, compared with the wild strain 2-10515. Differences in Colony morphology, cell growth, the rate of glucose consumption, pH, production of ethanol and total protein were assayed. The results showed that the colony morphology of Y518 was similar to that of 2-10515, but smaller and grew more slowly. In addition, no significant signs of growth were detected when Y518 was growing in the non-fermented carbon source. Therefore, Y518 was respiratory-deficient. Y518 had a lower rate of glucose consumption, maintained lower pH, produced less glycerol and kept high levels of protein expression. The energy metabolism of Y518 was analyzed by comparing with 2-10515. Previous studies showed that intracellular ROS level in Y518 increased significantly, thus Y518 suffered oxidative stress. Glycolytic enzymes are one of the main targets of ROS oxidation. The activities of the key enzymes involved in glycolytic pathway (hexokinase, phosphofructokinase and pyruvate kinase) were restrained and kept stable at the late fermentation. The intracellular content of product pyruvic acid was also III declining. Meanwhile the intracellular content of intermediate products of TCA cycle (citrite, succinite) decreased obviously in the fermentation process. Intracellular concertration of ATP and the activity of H+-ATPase of Y518