Bacillus sphaericus로부터 Cholesterol Oxidase의 정제 및 특성

Purification and Characterization of Cholesterol Oxidase from Bacillus sphaericus

  • 서형주 (고려대학교 병설 보건전문대학 식품영양과) ;
  • 김태웅 (강원대학교 생화학과) ;
  • 손흥수 (한림대학교 한국영양연구소)
  • 발행 : 1993.10.01

초록

The cholesterol oxidase produced from Bacillus sphaericus was purified and characterized. Through a series of purification procedures including DEAE-Toyoperal 650C, Sephadex G-200 and DEAE-Sephadex A-50 column chromatography, the purified enzyme was shown to have a specific activity of 0.179 units/mg protein having 31.8 fold purification and final yield of 12%. The molecular weight of the enzyme was estimated to be 47kDa and 47.tkDa by Sephadex G-200 chromatography and SDS-PAGE. The optimum temperature and pH for the enzyme were 30C and 6.0, respectively. The activity of the purified cholesterol oxidase was inhibited by Fe2+ and Hg+.

키워드

참고문헌

  1. Clin. Chem. v.20 Enzymatic determination of total serum cholesterol Allian,C.C.;L.S.Poon;W.Richmond;P.C.Fu
  2. Clin. Chem. v.29 Improved enzymatic determination of total cholesterol in tissues Cho,B.H.S.
  3. J. Agric. Food Chem. v.39 Reactvity of milk cholesterol with bacterial cholesterol oxidses Smite,M.;C.Sullivan;N.Goodmen
  4. Biochem. J. v.38 The microbiological degradation of steroids. 2. Oxidation of cholesterol by proactinomyces spp. Turfitt,G.E.
  5. Clin. Chem. v.19 Preparation and properties of bacterial cholesterol oxidase from Nocardia sp. and its application to the enzymatic assay of total cholesterol in serum Richmond,W.
  6. J. Biochem. v.79 Some enzymatic properties of 3β-hydroxsteroid oxidase produced by Streptomyces vilascens Tomioka,H.;Y.Kawakami;S.Nagamura
  7. Chem. Pharm. Bull. v.21 A method to screen anticholesterol substances produced by microbes and a new choleaterol oxidse produced by Streptomyces violascens Fukuda,H.;Y.Kawakami;S.Nagamura
  8. Agric. Biol. Chem. v.37 Isolation and crystallization of extracellular 3β-hydroxysteroid oxidase of Brevibacterium sterolicum nov. sp. Uwajima,T.;H.Yagi;S.Nagamura;O.Terada
  9. 特聞昭 54-105291 Cholesterol 酸化酵素製造法 松井條;中島和籃;谷口勉;大林見;田邊條;山田秀明
  10. Preparative Biochem. v.20 Production, purification and some properties of cholesterol oxidase from a Strepmyces sp. Lartillot,S.;P.Kedziora
  11. J. Int. Biotechnol. Korea Univ. v.4 Production of cholesterol oxidase by Bacillus sphaericus Son,H.S.
  12. Anal. Biochem. v.72 A rapid and sensitive method for the quantification of μg proteins Bradford,M.M.
  13. Biochem. J. v.96 The gel filtration behaviour of proteins milecularr weights over awide range Andrews,P.
  14. Nature v.227 Cleavege of Structural proteins during the assembly of head of Bacteriophage T4 Andrews,P.
  15. J. Bio. Chem. v.249 Resolution of bacterial proteins by polyacryamide gel electrophoresis on slabs Ames,G.F.L.
  16. 산업생물학회지 v.20 토양 미생물 HSL 613 이 생산하는 cholesterol oxidase의 정제 및 특성 이홍수;이승철;권태종;정태화
  17. Chem. Pharm. Bull. v.26 Purification of 3β-hydroxysteroid oxidase of Streptomyces violascens origen by affinity chromatography on cholesterol Kamei,T.;Y.Takiguchi;H.Suxuki;S.Nakamura
  18. J. Ferment. Technol. v.55 Purification and some proterties of an extracellular 3β-hydroxysteroid oxidase produced by Corynebcaterium cholesterolicum Shirokane,Y.;K.Nakamura;K.Mizusawa
  19. Appl. Microbiol. Biotechnol. v.31 Purification and characterization of cholesterol oxidase from Pseudomonas sp. and toxonomic study of the strain Lee,S.Y.;H.I.Rhee;B.K.Park
  20. J. Agric. Food Chem. v.37 Properties of the purified extracellular cholesterol oxidase from Rhodococcus equi No. 13 Watanabe,K.;H.Aihara;Y.Nakagawa;R.Nakamura;K.Susaki
  21. Agric. Biol. Chem. v.52 Purification and some properties of cholesterol oxidase producted by an inducible and a constitutive mutant Liu,W.;M.H.Meng;K.S.Chem
  22. Toyobo Enzyme Toyobo Co. Ltd.
  23. J. Biochem. v.85 Purification and some properties of cholesterol oxidase from Schicophyllum commume with covalently bound flavin Fukuyama,M.;Y.Miyake