Implementation of a transhydrogenase-like shunt to counter redox imbalance during xylose fermentation in Saccharomyces cerevisiae

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• 作者：Hiroyuki Suga (1)
  Fumio Matsuda (2) (3)
  Tomohisa Hasunuma (2)
  Jun Ishii (2)
  Akihiko Kondo (1) (3)
  
• 关键词：Transhydrogenase ; like shunt ; Malic enzyme ; Anaplerotic pathway ; Xylose fermentation ; Saccharomyces cerevisiae
• 刊名：Applied Microbiology and Biotechnology
• 出版年：2013
• 出版时间：February 2013
• 年：2013
• 卷：97
• 期：4
• 页码：1669-1678
• 全文大小：383KB
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• 作者单位：Hiroyuki Suga (1)
  Fumio Matsuda (2) (3)
  Tomohisa Hasunuma (2)
  Jun Ishii (2)
  Akihiko Kondo (1) (3)
  
  1. Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe, Hyogo, 657-8501, Japan
  2. Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe, Hyogo, 657-8501, Japan
  3. RIKEN Biomass Engineering Program, 1-7-22 Suehirocho, Tsurumi, Yokohama, 230-0045, Japan
  
• ISSN：1432-0614

文摘

Three enzymes responsible for the transhydrogenase-like shunt, including malic enzyme (encoded by MAE1), malate dehydrogenase (MDH2), and pyruvate carboxylase (PYC2), were overexpressed to regulate the redox state in xylose-fermenting recombinant Saccharomyces cerevisiae. The YPH499XU/MAE1 strain was constructed by overexpressing native Mae1p in the YPH499XU strain expressing xylose reductase and xylitol dehydrogenase from Scheffersomyces stipitis, and native xylulokinase. Analysis of the xylose fermentation profile under semi-anaerobic conditions revealed that the ethanol yield in the YPH499XU/MAE1 strain (0.38?±-.01?g g? xylose consumed) was improved from that of the control strain (0.31?±-.01?g g? xylose consumed). Reduced xylitol production was also observed in YPH499XU/MAE1, suggesting that the redox balance was altered by Mae1p overexpression. Analysis of intracellular metabolites showed that the redox imbalance during xylose fermentation was partly relieved in the transformant. The specific ethanol production rate in the YPH499XU/MAE1–MDH2 strain was 1.25-fold higher than that of YPH499XU/MAE1 due to the additional overexpression of Mdh2p, whereas the ethanol yield was identical to that of YPH499XU/MAE1. The specific xylose consumption rate was drastically increased in the YPH499XU/MAE1–MDH2–PYC2 strain. However, poor ethanol yield as well as increased production of xylitol was observed. These results demonstrate that the transhydrogenase function implemented in S. cerevisiae can regulate the redox state of yeast cells.