L-Erythrulose Production by Oxidative Fermentation is Catalyzed by PQQ-Containing Membrane-bound Dehydrogenase(Microbiology & Fermentation Technology)

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概要

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• Thermotolerant Gluconobacter frateurii CHM 43 was selected for L-erythrulose production from meso-erythritol at higher temperatures. Growing cells and the membrane fraction of the strain rapidly oxidized meso-erythritol to L-erythrulose irreversibly with almost 100% of recovery at 37℃. L-Erythrulose was also produced efficiently by the resting cells at 37℃ with 85% recovery. The enzyme responsible for meso-erythritol oxidation was found to be located in the cytoplasmic membrane of the organism. The EDTA-resolved enzyme required PQQ and Ca^<2+> for L-erythrulose formation, suggesting that the enzyme catalyzing meso-erythritol oxidation was a quinoprotein. Quinoprotein membrane-bound meso-erythritol dehydrogenase (QMEDH) was solubilized and purified to homogeneity. The purified enzyme showed a single band in SDS-PAGE of which the molecular mass corresponded to 80 kDa. The optimum pH of QMEDH was found at pH 5.0. The Michaelis constant of the enzyme was found to be 25 mM for meso-erythritol as the substrate. QMEDH showed a broad substrate specificity toward C3-C6 sugar alcohols in which the erythro form of two hydroxy groups existed adjacent to a primary alcohol group. On the other hand, the cytosolic NAD-denpendent meso-erythritol dehydrogenase (CMEDH) of the same organism was purified to a crystalline state. CMEDH showed a molecular mass of 60 kDa composed of two identical subunits, and an apparent sedimentation constant was 3.6 s. CMEDH catalyzed oxidoreduction between meso-erythritol and L-erythrulose. The oxidation reaction was observed to be reversible in the presence of NAD at alkaline pHs such as 9.0-10.5. L-Erythrulose reduction was found at pH 6.0 with NADH as coenzyme. Judging from the catalytic properties, the NAD-dependent enzyme in the cytosolic fraction was regarded as a typical pentitol dehydrogenase of NAD-dependent and the enzyme was independent of the oxidative fermentation of L-erythrulose production.

• 2002-02-23

著者

• 足立 収生

  山口大学農学部生物機能科学科

• 足立 収生

  山口大・農・生物機能

• 品川 恵美子

  宇部高専物質工学科

• Moonmangmee Duangtip

  山口大農応用生物化学

• Theeragool G.

  カセサート大学理

• MOONMANGMEE Duangtip

  Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University

• TOYAMA Hirohide

  Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University

• THEERAGOOL Gunjana

  Department of Microbiology, Faculty of Science, Kasetsart University

• LOTONG Napha

  Department of Microbiology, Faculty of Science, Kasetsart University

• MATSUSHITA Kazunobu

  Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University

• ADACHI Osao

  Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University

• SHINAGAWA Emiko

  Department of Chemical and biological Engineering, Ube National College of Technology

• Toyama Hirohide

  Department Of Biological Chemistry Faculty Of Agriculture Yamaguchi University

• Adachi Osao

  Derartment Of Blological Chemistry Faculty Of Agriculture Yamaguchi University

• Adachi Osao

  Department Of Agricultural Chemistry Yamaguchi University

• Lotong Napha

  Department Of Microbiology Faculty Of Science Kasetsart University

• Lotong Napha

  カセサート大理 微生物

• Shinagawa Emiko

  Department Of Chemical And Biological Engineering Ube National College Of Technology

• Theeragool G.

  カセサート大理微生物

• Theeragool Gunjana

  カセサート大理 微生物

• Matsushita Kazunobu

  Department Of Biological Chemistry Faculty Of Agriculture Yamaguchi University

• Matsushita Kazunobu

  Derartment Of Blological Chemistry Faculty Of Agriculture Yamaguchi University

• Moonmangmee Duangtip

  Department Of Microbiology Faculty Of Science King Mongkut's University Of Technology Thonburi

• Moonmangmee Duangtip

  山口大農 生物機能科学

• 足立 収生

  山口大学 農

• 足立 収生

  山口大・農化

• Matsushita K

  Department Of Biological Chemistry Faculty Of Agriculture Yamaguchi University

• Theeragool Gunjana

  Department Of Microbiology Faculty Of Science Kasetsart University

• 松下 一信

  山口大 農

• Lotong Napha

  Department Of Microbioloby Faculty Of Science Kasetsart University

• Shinagawa Emiko

  Department of Agricultural Chemistry, Faculty of Agriculture, Yamaguchi University

• Matsushita Kazunobu

  Department of Agricultural Chemistry, Faculty of Agriculture, Yamaguchi University

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