Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Study on the Production and the Culture Condition of Cholesterol Oxidase from Bacillus megterium SFO41

Bacillus megaterium SFO41에 의한 Cholesterol Oxidase의 생산 및 최적 배양 조건

  • 김관필 (경북대학교 식품공학과) ;
  • 이창호 (경북대학교 식품공학과) ;
  • 우철주 (경북대학교 식품공학과) ;
  • 박희동 (경북대학교 식품공학과)
  • Published : 2001.06.01
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Abstract

A novel strain of SFO41 producing a large amount of cholesterol oxidase as an extracellular enzyme isolate from Korean salt fermented foods. The strain was identified as Bacillus megaterium based on morphological, cultural and physiological characteristics. Experiments were carried out to optimized the condition of cholesterol oxidase production using B. megaterium SFO41. B. megaterium SFO41 was shown to give the maximum yield of cholesterol oxidase in the medium containing 2.0% glucose, 0.5% yeast extract. 0.03% $MgSO_4{\cdot}7H_2O,\;0.02%\;K_2HPO_4,\;0.2%\;NH_4NO_3$ and 0.2% cholesterol. The optimum culture conditions, temperature, initial pH and agitation speed were $30^{\circ}C$, 7.0 and 150 rpm, respectively. The enzyme production reached a maximum level at 24 hr of cultivation (2.37 U).

우리나라 전통 발효식품(침채류 및 젓갈류)으로부터 cholesterol oxidase 생산성이 있는 균주를 분리하고 이들 분리된 균주들로부터 여러 단계의 균주 선멸시험을 통하여 cholesterol oxidase 생산성이 우수한 미생물을 선별하여 그 특성을 조사하였다. Cholesterol oxidase의 생산성이 가장 우수한 SF041의 형태학적, 배양학적 및 생리학적 특성을 조사하여 Bergey's Mannual of Systematic Bacteriology의 분류기준에 따라 동정한 결과 Bacillus megaterium 또는 그 유연균으로 동정되어 분리균을 Bacillus megaterium SFO41로 명명하였다. Cholesterol oxidase의 생산 조건을 검토한 결과 최적 배지 조성은 2.0% glucose, 0.5% yeast extact, 0.03% $MgSO_4\;7H_2O,\;0.02%\;K_2HPO_4,\;0.2%\;NH_4NO_3$, 0.2% cholesterol로 판명되었으며, 배영 조건은 $30^{\circ}C$, 초기 pH 7.0, 진탕 속도는 150 rpm에서 24시간 배양 시 효소 생성이 가장 우수하였다(2.37 U).

Keywords

References

  1. Clin Chem. v.20 Enzymatic determination of total serum cholesterol Allina, C C.;Poon, L S.;Richmond, W.;Fu, P.C
  2. Clin. Chem. v.29 Improved enzymatic determination of total cholesterol in tissues Cho, B.H.S.
  3. Ann Clin. Biochem. v.10 An investigation of the determination of serum cholesterol by enzymatic method Flegg, H.M.
  4. Clin Chem v.28 Enzymatic determination of free and esterified cholesterol in serum by microcalonmetry Rehak, N.N.;Young, D.S
  5. Clin. Chem v.29 Reagent for the enzymatic determination of serum total cholesterol with improved lipolytic efficiency Sidel, J;Hagele, E O.;Ziegenhorn, J.;Wahlefeld, A.W.
  6. J. Chromatogr v.101 Application of cholesterol oxidase in the analysis of steroids Smith, A G.;Brooks, C.J.W
  7. 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.
  8. Am. J. Med. v.59 Accidental hypothermia in alcoholic population Weyman, A.E.
  9. Biochem. J. v.38 The microbiological degradation of steroids 2 Oxidation of cholesterol by Proactinomyces spp Turfitt, G.E.
  10. J. Bacteriol. v.58 The ability of soil microorganisms the decompose steroids Schatz, A.;Savard, K;Pinter, I.J.
  11. J. Biol. Chem v.205 Purification and properties of a β-hydroxysteroid dehydrogenase Talalay, P.;Dobson, M M
  12. J. Biol. Chem. v.206 Studies on the microbiol degradation of cholesterol Stadtman, T.C.;Cherkes, A.;Anfinsen, C.B.
  13. Biotechnol Bioeng. v.18 The kinetics of cholesterol oxidase synthesis by Nocardia rhodocruos Buckland, B C.;Lilly, M D.;Dunnill, P.
  14. J. Amer Chem. Soc v.87 Mechanism of steroid oxidation by microorganism Sih, C.J;Wang, K.C.
  15. J. Bacteriol. v.54 Microbiological agencies in the degradation of steroids Turfitt, G.E
  16. Agric. Biol. Chem. v.22 Microbiol transformation of steroids, Ⅰ.Decomposition fo cholesterol by microorganisms Arima, K.;Nagasawa, M.;Bae, M.;Tamura, G
  17. Chem. Pharm. Bull (Tokyo) v.26 Purification of 3β-hydroxy steroid oxidase of Streptomyces violascens orgin by affinity chromatography on cholesterol Kamei, T;Takiguchi, Y.;Suzuki, H.;Nakamura, S
  18. J. Biol. Chem. v.79 Some enzymatic properties of 3β-hydroxysteroid oxidase of Streptomyces violascens Tomioka, H.;Kagawa, M.;Nagamura, S
  19. Agric. Biol. Chem v.37 Isolation and crystallization of extracellular 3β-hydroxysteroid oxidase of Brevibacterium sterolicum nov. sp. Uwajima, T.;Yagi, H;Nagamura, S.;Terade, O.
  20. Agric. Biol Chem. v.38 Properties of crystalline 3β-hydroxysteroid oxidase of Brevibacterium sterolicum Uwajima, T.;Yagi, H.;Terada, O.
  21. J. Ferment. Technol. v.55 Purification and some properties of an extracellular 3 β-hydroxysteroid oxidase produced by Corynebacterium cholesterolicum Shirokane, Y.;Nakamura, K.;Mizusawa, K.
  22. J. Biotechnol. v.33 Membrane-bound cholesterol oxidase from Rhodococcus sp cells production and extraction Krent. J.;Lefebvre, G.;Germain, P.
  23. J. Agric. Food Chem. v.37 Properties of the purified extracellular cholesterol oxidase from Rhodococcus equi No. 23 Watanabe, K.;Aihara, H.;Nakagawa, Y.;Sasaki, T.
  24. J. Gem. Appl. Microbiol. v.32 Isolation and identification of cholesterol-degradation Rhodococcus strainns food of animal orgin and their cholesterol oxidase activities Watanabe, K.;Shimizu, H;Aihara, H;Nakamura, R.;Suzuki, K I;Momagata, K.
  25. Chem. Pharm. Bull. v.21 A method to screen anticholesterol substances produced by microbes and a new cholesterol oxidase produced by Streptomyces violascens Fukuda, H.;Kawakami, Y.;Nagamura, S.
  26. Kor. J. Appl. Microbiol. Bioeng v.16 Microbiol conversion of cholesterol to 4-androstene-3.17-dione by intermittent addition of substrate Choi, S.K.;Kim, H S.;Park, Y.H.
  27. Appl. Microbiol. Biotechnol v.31 Purification and characterization of cholesterol oxidase from Pseudomonas sp. and taxonomic study of the strain Lee, S.Y.;Rhee, H I;Tae, W.C.;Shin, J.C.;Park, B.K
  28. Kor. J. Appl. Microbiol. Bioeng. v.20 Studies on the isolation of cholesterol oxidase producing soil microorganism and the culture condition for the production of high activity cholesterol oxidase Lee, I.A.;Choe, Y.K.;Lee, H.S;Choe, I S.;Chung, T.W.
  29. Kor J Biotechnol. Bioeng. v.13 Purification and characterization of cholesterol oxidase produced by Rhodococcus sp. 3T6-5MJ isolated from Changran-jeot Park, S.H.;Kim, H.S.;Lee, Y.S.;Kwon, I.B.;Chun, U H.
  30. J. Appl. Bacteriol. v.59 Biotransformation of sugar-cane sterols into Androstal, 4-dione-3,17-dione (ADD) by Arthrobacter globiformis and Str. Oxidans Srivastava, S K.;Srivastava, R.A.K.;Mathur, S N
  31. Enzyme. Microb. Technol. v.14 Formation of mixed crystals in microbiol conversion of sterols and steroids Goetschel, R;Bar. R.
  32. J. Food Sci. v.49 Effect of consuming yogurt prepared with three culture strains on human serum lipoproteins Jaspers, D.A.;Massey, L.K.;Luedecke, L.O.
  33. Bergey's Manual of Determinative Bateriology(9th ed.) John, G.H.;Noel, R.K;Peter, H.S.S.;James, T S;Stanely, T.W.
  34. Proc. Natl. Sci. v.7 Production of cholesterol oxidase by antibiotic resistant mutant and a constitutive mutant Arthrobacter simplex B-7 Liu, W.;Hsu, J;Wang, W
  35. Agric. Biol. Chem. v.5 Purification and some properties of cholesterol oxidase produced by an inducible and a constitutive mutant Arthrobacter simplex Liu, W.;Meng, M.;Chen, K.
  36. J Microbiol Biotechnol. v.7 Microbiol transformation of sterols to C19-steroids by Rhodococcus equi Ahmad, S.;Roy, P K.
  37. Kor. J. Appl Microbiol. Biotechnol. v.21 Purification and characterization of cholesterol oxidase from Bacillus sphaericus Suh, H J;Kim, T.W.;Son, H.S.