Characteristics and Action Pattern of Pretense from Bacillus subtilis CCKS-111 in Korean Traditional Soy Sauce

한국재래간장으로 부터 분리한 Bacillus subtilis CCKS-111이 생성하는 Protease의 특성 및 작용양상

  • Choi, Cheong (Dept. of Food Science and Technology, Yeungnam University) ;
  • Choi, Kwang-Soo (Dept. of Food Science and Technology, Yeungnam University) ;
  • Cho, Young-Je (Dept. of Food Engineering, Sangju National Polytechnic University) ;
  • Lim, Sung-il (Dept. of Food Science and Technology, Yeungnam University) ;
  • Kim, Sung (Dept. of Food Science and Technology, Yeungnam University) ;
  • Son, Jun-Ho (Dept. of Food Science and Technology, Yeungnam University) ;
  • Lee, Hee-Duck (Dept. of Food Science and Technology, Yeungnam University) ;
  • Kim, Young-Hwal (Dept. of Clinical Pathology, Daegu Junior College)
  • 최청 (영남대학교 식품가공학과) ;
  • 최광수 (영남대학교 식품가공학과) ;
  • 조영제 (상주산업대학교 식품공학과) ;
  • 임성일 (영남대학교 식품가공학과) ;
  • 김성 (영남대학교 식품가공학과) ;
  • 손준호 (영남대학교 식품가공학과) ;
  • 이희덕 (영남대학교 식품가공학과) ;
  • 김영활 (대구보건전문대학 임상병리과)
  • Published : 1996.12.01

Abstract

An alkaline pretense Producing microorganism was isolated from Korean traditional soy sauce and identified as Bacillus subtilis CCKS-111. The optimum culture condition of Bacillus subtilis CCKS-111 for the production of alkaline pretense was as follow: 2% soluble starch, 0.2% peptone, 0.1% (NB$_4$)$_2$S$_2$O$_{8}$ , 0.2% MgSO$_4$, pH 7.0, 35$^{\circ}C$ and 24hrs. The optimum pH and temperature for the enzyme activity of alkaline pretense producing Bacillus subtilis CCKS-111 were pH 9.0 and 5$0^{\circ}C$, respectively. The enzyme was relatively stable at pH 6.0~11.0 and at temperature below 4$0^{\circ}C$. The activity of the enzyme was inhibited by $K^{+}$ and Hg$^{2+}$, whereas Cu$^{2+}$ exhibited rather activating effects on the enzyme activity. Ethylenediaminetetraacetic acid and phenylmethanesulfonyl fluoride inhibited the enzyme activity. This indicates that this is serine pretense which requires metal ion group for the enzyme activity. Km value was 2.313$\times$10$^{-4}$ M/L, V$_{max}$ value was 39.216$\mu\textrm{g}$/min. This enzyme hydrolyzed casein more rapidly than the hemoglobin.lobin.

한국재래간장으로부터 분리한 Bacillus subtilis CCKS-111이 생성하는 protease 생산의 최적 배양조건은 2% soluble starch, 0.2% peptone, 0.1%(MH$_4$)$_2$S$_2$O$_{8}$ , 0.2% MgSO$_4$, pH 7.0, 35$^{\circ}C$ 에서 24시간 배양했을 때이다. 효소의 최적 작용 pH와 온도는 pH 9.0, 5$0^{\circ}C$였으며 , pH 6.0~11.0의 범위와 5$0^{\circ}C$이상에서 불안정하였다. 금속이온 중 Cu$^{2+}$에 의하여 활성이 증대되었으나 $K^{2+}$, Hg$^{2+}$등에 의하여 효소활성이 저해되었다. Ethylenediamineteraacetic acid 와 phenylmethane sulfonyl fluoride 처리에 의해 활성이 저해되어 금속이온의 영향을 받는 serine protease로 확인되었다. Km값은 2.313$\times$$10^{-4}$ M, V$_{max}$ 값은 39.216$\mu\textrm{g}$/min이었으며, hemoglobin보다 casein을 더 잘 가수분해하였다.

Keywords

References

  1. Process Biochemistry v.1 Fish protein hydrolysates Im, M.
  2. Nippon Shokuhin Kogyo Gakkaishi v.36 Effect of protease on textural properties of wheat flour dough Ishide, K. I.;Nagasaki, M.
  3. Agric. Biol. Chem. v.35 Production of alkaline enzyme by alkalophilic microorganism Horikoshi, K.
  4. Alkalophilic microorganisms Horikoshi, K.;Aakiba, T.
  5. Biochemica et Biophysica. Acta v.896 Specificites of extracellular and ribosomal serine proteinases from Bacillus natto a food microorganism Ichishima, E.;Takada, V.;Taira, K.;Takeuchi, M.
  6. Agric. Biol. Chem. v.30 Studies on bacterial proteinase part 16. Purification. Crystalization and some enzymatic properties of alkaline protease of Bacillus subtilis var. amylosacchariticus Tsuru, D.;Heizokira, K.;Yamamoto, T.
  7. Yakugaku Zasshi v.88 Studies on Aspergillus proteinase Ito, M.;Sugiura, M.
  8. Agric. Biol. Chem. v.36 Enzymatic properties of alkaline proteinase from A. candidus Tadahiko, O.;Seiichi, N.
  9. Agric. Biol. Chem. v.42 Purification and characterization of crystalline microbial alkaline proteinase inhibitors produced by Streptomyces WT-27 Takashi, W.;Sawao, M.
  10. J. Ferment. Technol. v.54 Alkaline proteinase production from methyl acetate by alkalophilic Bacillus sp Makio, K.;Koki, H.
  11. J. Furment. Technol. v.63 Purification and some properties of serine proteinase from a mutant fo A. niger Kazuo, S.;Kyo, S.;Kinichi, M.
  12. 한국영양식량학회지 v.18 Aspergillus fumigatus에 의한 alkaline protease의 생산과 정제 차원섭;조영제;최청
  13. 한국산업미생물학회지 v.17 Streptomyces sp. YSA-130이 생성하는 alkaline protease의 정제와 특성 윤성우;익강표;윤주현;신철수;오두한
  14. 한국농화학회지 v.31 Bacillus subtilis가 생산하는 alkaline protease에 관한 연구 장신재;김윤숙;성하진;최용준;양한철
  15. 한국산업미생물학회지 v.18 호알칼리성 Bacillus속 균주가 생산하는 alkaline protease의 특성 김태호;박성희;이동선;권택규;김종국;홍순덕
  16. 한국영양식량학회지 v.20 Bacillus LY-353이 생산하는 protease의 정제 및 특성 이병우;유영선;임금형;최춘언
  17. 한국산업미생물학회지 v.23 Bacillus sp. KUN-17 균주가 생산하는 균체의 serine protease의 정제 및 특성 황세영
  18. 한국산업미생물학회지 v.18 Bacillus sp.가 생산하는 호알칼리성 protease의 부분정제 및 특성 안장우;오태광;박용하;박관화
  19. 酵素硏究法 v.Ⅱ Haginhara, B.
  20. J. Biol. Chem. v.193 Protein measurement with the folin phenol reagent Lowry, O. H.;Rosebrough, N. J.;Farr, L. A.;Randal, R. J.
  21. Korean Biochem. J. v.27 Purification and characterization of an extraxellular alkaline protease from Bacillus subtilis RM615 Moon, S. Y.;Oh, T. K.;Rho, H. M.
  22. Kor. J. Appl. Microbiol. Biotechnol. v.19 Screening of Bacillus sp. M-71 with high alkaline protease productivity and some properties of the enzyme Oh, S. H.;Oh, P. S.
  23. 한국영양식량학회지 v.20 Bacillus subtilis LY-353이 생산하는 protease의 정제 및 특성 이병우;유영선;임근형;최춘언
  24. 한국산업미생물학회지 v.23 고온성 Bacillus amylolique faciens NS 15-4가 생산하는 내열성 protease의 특성 김형권;김기현;이정기;김영옥;남희섭;오태광
  25. 한국산업미생물학회지 v.20 Streptomyces griseus HC-1141이 생산하는 alkaline protease의 특성 및 작용양상 최청;정영건;성삼경;최광수;이재성;조영제;천성숙
  26. 한국영양식량학회지 v.18 Aspergillus fumigatus가 생산하는 alkaline protease의 특성과 작용양상 차원섭;최청
  27. 한국생화학회지 v.24 Bacillus sp. CW-1121이 생성하는 alkaline protease의 특성 및 작용양상 이우제;조영제;손규목;최청