Purification and Characterization of Intracellular and Extracellular, Thermostable and Alkali-Tolerant Alcohol Oxidases Produced by a Thermophilic Fungus, Thermoascus aurantiacus NBRC 31693(Enzymology, Protein Engineering, and Enzyme Technology)

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概要

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• Intracellular and extracellular alcohol oxidases (AOint and AOext) were purified from the liquid and solid cultures of a thermophilic fungus, Thermoascus aurantiacus NBRC 31693, as electrophoretically and isoelectrophoretically homogeneous proteins, respectively. Both enzymes contained a flavin adenine dinucleotide (FAD) cofactor and were stained with Schiff's reagent. The molecular weight of AOint was estimated to be about 320kDa and its subunit was 75kDa. The molecular weight of AOext was about 560kDa, and it was composed of two types of subunits (75kDa and 59kDa). The pIs of AOint and AOext were 5.88 and 6.08, respectively. AOint and AOext were stable up to 60℃ and 55℃, respectively. The enzymes were stable over a wide range of pH from 6 to 11. AOint oxidized short straight-chain alcohols (K_m for methanol, 13.5mM and K_m for ethanol, 15.8mM). On the other hand, AOext could oxidize secondary alcohols and aromatic alcohols (veratryl alcohol and benzyl alcohol) in addition to straight-chain alcohols (K_m for methanol, 0.5mM and K_m for ethanol, 10.2mM).

• 社団法人日本生物工学会の論文
• 2005-04-25

著者

• OHNO Nobuko

  Department of Medicine, University of Pennsylvania School of Medicine

• Fujii T

  Kyoto Univ. Kyoto

• Fujii T

  Department Of Applied Biological Chemistry The University Of Tokyo

• YAMAGUCHI Yube

  Research and Education Center for Genetic Information, Nara Institute of Science and Technology

• KO HEE

  Department of Bioresources Science, Graduate School of Science and Technology, Chiba University

• YOKOYAMA YUICHI

  Department of Bioresources Science, Graduate School of Science and Technology, Chiba University

• AMACHI SEIGO

  Department of Bioresources Science, Graduate School of Science and Technology, Chiba University

• SHINOYAMA HIROFUMI

  Department of Bioresources Science, Graduate School of Science and Technology, Chiba University

• FUJII TAKAAKI

  Department of Bioresources Science, Graduate School of Science and Technology, Chiba University

• Ohno Nobuko

  Department Of Health And Nutrition School Of Home Economics Wayo Woman's University

• Fujii Takeshi

  National Institute for Agro-Environmental Sciences

• Yamaguchi Y

  Department Of Bioresources Science Graduate School Of Science And Technology Chiba University

• Yamaguchi Yoshitaka

  Department Of Biotechnology Faculty Of Engineering Fukuyama University

• Fujii T

  Inst. Biol. Sci. Univ. Tsukuba

• Amachi Seigo

  Div. Of Bioresource Sci. Graduate School Of Horticulture Chiba Univ.

• Ohno Nobuhiro

  Department Of Health And Nutrition School Of Home Economics Wayo Woman's University

• Yamaguchi Y

  Research And Education Center For Genetic Information Nara Institute Of Science And Technology

• Ko Hee

  Department Of Bioresources Science Graduate School Of Science And Technology Chiba University

• Fujii T

  Div. Of Bioresource Sci. Graduate School Of Horticulture Chiba Univ.

• OKADOME MIHO

  Department of Health and Nutrition, School of Home Economics, Wayo Woman's University

• Okadome Miho

  Department Of Health And Nutrition School Of Home Economics Wayo Woman's University

• Shinoyama H

  Department Of Advanced Bioresources Science Graduate School Of Science And Technology Chiba Universi

• Shinoyama Hirofumi

  Department Of Agricultural Chemistry Chiba University

• Fujii Takaaki

  Department Of Biological Chemistry Faculty Of Agriculture Yamaguchi University

• Yokoyama Yuichi

  Department Of Applied Chemistry Faculty Of Engineering Osaka Institute Of Technology

• Ko Hee-Sun

  Department of Bioresources Science, Graduate School of Science and Technology, Chiba University

• FUJII Takaaki

  Department of Agricultural Chemistry, Chiba University

• Shinoyama Hirofumi

  Department of Advanced Bioresources Science, Graduate School of Science and Technology, Chiba University

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