Distinct Roles of Two Anaplerotic Pathways in Glutamate Production Induced by Biotin Limitation in Corynebacterium glutamicum(GENETICS, MOLECULAR BIOLOGY, AND GENE ENGINEERING)

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概要

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• Corynebacterium glutamicum is a biotin auxotrophic bacterium in which glutamate production is induced under biotin-limited conditions. During glutamate production, anaplerotic reactions catalyzed by phosphoenolpyruvate carboxylase (PEPC) and a biotin-containing enzyme pyruvate carboxylase (PC) are believed to play an important role in supplying oxaloacetate in the tricarboxylic acid cycle. To understand the distinct roles of PEPC and PC on glutamate production by C. glutamicum, we observed glutamate production induced under biotin-limited conditions in the disruptants of the genes encoding PEPC (ppc) and PC (pyc), respectively. The pyc disruptant retained the ability to produce high amounts of glutamate, and lactate was simultaneously produced probably due to the increased intracellular pyruvate levels. On the other hand, the ppc knockout mutant could not produce glutamate. Additionally, glutamate production in the pyc disruptant was enhanced by overexpression of ppc rather than disruption of the lactate dehydrogenase gene (ldh), which is involved in lactate production. Metabolic flux analysis based on the ^<13>C-labeling experiment and measurement of ^<13>C-enrichment in glutamate using nuclear magnetic resonance spectroscopy revealed that the flux for anaplerotic reactions in the pyc disruptant was lower than that in the wild type, concomitantly increasing the flux for lactate formation. Moreover, overexpression of ppc increased this flux in both the pyc disruptant and the wild type. Our results suggest that the PEPC-catalyzed anaplerotic reaction is necessary for glutamate production induced under biotin-limited conditions, because PC is not active during glutamate production, and overexpression of ppc effectively enhances glutamate production under biotin-limited conditions.

• 社団法人日本生物工学会の論文
• 2008-07-25

著者

• Wachi Masaaki

  Department of Bioengineering, Tokyo Institute of Technology

• Hirasawa Takashi

  Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osak

• Shimizu Hiroshi

  Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osak

• Hirasawa Takashi

  Graduate School Of Information Science And Technology Osaka University

• Hirasawa Takashi

  Department Of Bioengineering Tokyo Institute Of Technology

• Hirasawa Takashi

  Osaka University

• SATO Hiroki

  Department of Health Chemistry, Hoshi University

• NAGAHISA KEISUKE

  Department of Biotechnology, Graduate School of Engineering, Osaka University

• Orishimo Keita

  Department of Bioengineering, Tokyo Institute of Technology

• Shirai Tomokazu

  Department of Biotechnology, Graduate School of Engineering, Osaka University

• Orishimo Keita

  Department Of Bioengineering Tokyo Institute Of Technology

• Shirai Tomokazu

  Department Of Biotechnology Graduate School Of Engineering Osaka University

• Nagahisa Keisuke

  Department Of Bioinformatic Engineering Graduate School Of Information Science And Technology Osaka

• Nagahisa Keisuke

  Department Of Bioinformatic Engineering Graduate School Of Information Science And Technology Osaka

• Shimizu Hiroshi

  Department Of Bioinformatic Engineering Graduate School Of Information Science And Technology Osaka

• Wachi Masaaki

  Department Of Bioengineering Graduate School Of Bioscience And Biotechnology Tokyo Institute Of Tech

• Sato Hiroki

  Department Of Bioengineering Tokyo Institute Of Technology

• Nagahisa Keisuke

  Department Of Bioinformatic Engineering Graduate School Of Information Science And Technology Osaka

• Shimizu Hiroshi

  Department Of Animal Science Faculty Of Agriculture Hokkaido University

• Shimizu Hiroshi

  Department Of Bioinformatic Engineering Graduate School Of Information Science And Technology Osaka

• SHIMIZU HIROSHI

  Department of Anatomy, School of Medicine, Tokushima University

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