Purification and Some Properties of Cholesterol Oxidase Stable in Detergents from γ-Proteobacterium Y-134

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概要

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• Cholesterol oxidase (CHO) with high stability in detergents was found from an isolated strain, Y-134, belonging to the γ-subclass of Proteobacteria. CHO production reached its maximum by incubation at 30℃ for 12d. It was purified from cell-free extract prepared by mixing the cells with 0.4% Triton X-100. The absorption spectrum of the purified enzyme exhibited maxima at 274 and 410 nm, and a shoulder at 330 nm. The molecular mass was 115 kDa with two identical subunits of 58 kDa. The enzyme oxidized cholest-5-en-3β-ol (cholesterol) and 5α-cholestan-3β-ol (dihydro-cholesterol) at a high reaction rate, and the K_m value for cholesterol was 65 μM. The stability of the enzyme was higher than other CHOs in nonionic detergents with high values of hydrophile lipophile balance (HLB) such as Triton X-450 and sodium cholate. NH_2-terminal sequence analysis showed a high similarity to CHO from Burkholderia cepacia, but not to CHOs from Streptomyces or Brevibacterium.

• 社団法人日本生物工学会の論文
• 2003-03-25

著者

• Isobe Kimiyasu

  Department of Agro-bioscience, Faculty of Agriculture, Iwate University

• Isobe K

  Department Of Agro-bioscience Faculty Of Agriculture Iwate University

• Isobe Kimiyasu

  Department Of Agro-bioscience Faculty Of Agriculture Iwate University

• SHOJI KAYAKO

  Department of Agro-bioscience, Faculty of Agriculture, Iwate University

• NAKANISHI YUJI

  Gifu Research and Development Center, Amano Enzyme Inc.

• YOKOE MASAAKI

  Gifu Research and Development Center, Amano Enzyme Inc.

• WAKAO NORIO

  Department of Agro-bioscience, Faculty of Agriculture, Iwate University

• Nakanishi Yasuaki

  Department Of Fermentation Technology Faculty Of Engineering Hiroshima University

• Wakao Norio

  Department Of Agro-bioscience Faculty Of Agriculture Iwate University

• Wakao N

  Department Of Agro-bioscience Faculty Of Agriculture Iwate University

• Shoji Kayako

  Department Of Agro-bioscience Faculty Of Agriculture Iwate University

• Yokoe Masaaki

  Gifu Research And Development Center Amano Enzyme Inc.

• Nakanishi Yuji

  Gifu Research And Development Center Amano Enzyme Inc.

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