본문 바로가기
KERD
논문KCI 등재

청국장에서 분리된 Bacillus licheniformis의 Protease 생산을 위한 배지 최적화

Medium Optimization for the Protease Production by Bacillus licheniformis Isolated from Cheongkookjang.

1

조회

0

다운로드

14

인용

0

참고문헌

키워드

주제분류생물학

초록

청국장의 기호도를 개선하는 발효균으로 분리된 protease 생산균은 Bacillus licheniformis로 확인되었다. Protease 생산을 위한 배지를 최적화하기 위한 탄소원, 질소원, 인, 금속이온의 성분을 변화시키면서 균의 성장과 효소 생산성을비교하였다. Glucose를 탄소원으로 사용하였을 때는 균의 성장은 정상적으로 일어나지만, protease 생산이 심하게 억제되는 것으로 나타났으며, Potato starch를 탄소원으로 사용하였을 때 효소 생산성이 가장 높았다. 질소원으로는 yeast extact가 효소 생산에 가장 적합하였다. 한편 2가 금속이온중 Zn2+, Cu2+, Co2+을 배지에 첨가하였을 때는 균의 성장이 심하게 저해되었으며, 효소 생산도 되지 않았다. CaCl2를 첨가한 배지에서는 균이 성장과 효소 생산성이 증가되었다. Potato starch(1.5%), yeast extract(1.5%), CaCl2(0.7%),K2HPO4(0.03%)와 KH2PO4(0.03%)를 포함하는 것으로 구성된 최적화 배지에서 최대효소 생산성은 800 U/mL로 나타났으며 28시간 이후에는 배양액내 효소활성이 서서히 감소하였다.

Abstract

Bacillus licheniformis fermenting soybean product with highest score in consumer acceptance had been isolated from homemade Cheongkookjang. In order to develop the medium composition, effects of ingredients including nitrogen sources, carbon sources, metal ions and phosphate were examined for protease production of the isolate. Potato starch increased the protease productivity, while glucose repressed it. Yeast extract was the most effective nitrogen source for enzyme production. The calcium was found to increase protease activity slightly while cell growth and enzyme production was completely inhibited by divalent ions such as Zn2+, Cu2+ and Co2+. The maximum protease productivity was reached approximately 800 unit/mL in the optimized medium consisting of potato starch (1.5%), yeast extract (1.5%),CaCl2 (0.7%), K2HPO4 (0.03%) and KH2PO4 (0.03%). The protease activity of culture filtrate was gradually decreased after incubation for 28 h.

참고문헌 (25)

  1. Abdel-Fattah Y. R., "Bioprocess Development for Production of Alkaline Protease by Bacillus pseudofirmus Mn6 Through Statistical Experimental Designs", Journal of Microbiology and Biotechnology, 19(4), pp.378~386 (2009), 한국미생물·생명공학회
  2. Agrebi, R, "Fibrinolytic serine protease isolation from Bacillus amyloliquefaciens An6 grown on Mirabilis jalapa tuber powders", Appl. Biochem. Biotechnol., 162, pp.75~88 (2010)
  3. Fakhfakh, N, "Production and biochemical and molecular characterization of a keratinolytic serine protease from chicken featherdegrading Bacillus licheniformis RPk", Can. J. Microbiol., 55, pp.427~436 (2009)
  4. Fakhfakh-Zouari, N, "Application of statistical experimental design for optimization of keratinases production by Bacillus pumilus A1 grown on chicken feather and some biochemical properties", Process Biochem., 45, pp.617~626 (2010)
  5. Ferrero, M. A, "Thermostable alkaline protease Bacillus licheniformis MIR 29: isolation, production and characterization", Appl. Microbiol. Biotechnol, 45, pp.327~332 (1996)
  6. Gupta, R, "Bacterial alkaline proteases: molecular approaches and industrial applications", Appl. Microbiol Biotechnol, 59, pp.13~32 (2002)
  7. Haddar, A, "Low-cost fermentation medium for alkaline protease production by Bacillus mojavensis A21 using hulled grain of wheat and sardinella peptone", J. Biosci. Bioeng., 110, pp.288~294 (2010)
  8. Hadj-Ali, N. E, "Biochemical and molecular characterization of a detergent stable alkaline serine-protease from a newly isolated Bacillus licheniformis NH1", Enzyme Microb. Technol., 40, pp.515~523 (2007)
  9. 홍성욱, "면역증강물질 강화 청국장 발효", 한국식품과학회지, 38(4), pp.548~553 (2006), 한국식품과학회
  10. 현광욱, "재래식 된장으로부터 혈전용해활성을 나타내는 세균의 분리 및 동정", 한국미생물·생명공학회지, 33(1), pp.24~28 (2005), 한국미생물·생명공학회
  11. 김사숙, "청국장으로부터 분리한 Bacillus amyloliquefaciens D4-7이 분비하는 혈전용해효소의 특성 및 열안정성에 미치는 첨가물의 효과", 한국미생물·생명공학회지, 31(3), pp.271~276 (2003), 한국미생물·생명공학회
  12. 권하영, "청국장으로부터 면역증강활성이 우수한Bacillus pumilus JB-1의 뷴리 및 분리균의청국장 발효특성", 한국미생물·생명공학회지, 32(4), pp.291~296 (2004), 한국미생물·생명공학회
  13. 이동근, "청국장으로부터 Fibrin 분해 세균의 분리 및이를 이용한 발효 청국장의 생리활성", 한국미생물·생명공학회지, 34(4), pp.299~305 (2006), 한국미생물·생명공학회
  14. Gyu Hee Lee, "Sensory Comparison of Fermented Soybeans (Cheongkookjang) Inoculated with Various Bacillus species by Principal Component Analysis", Journal of the Korean Society for Applied Biological Chemistry, 52(6), pp.726~730 (2009), 한국응용생명화학회
  15. Lee, J, "Kong, and S. Park. 1999. Production of a fibrinolytic enzyme in bioreactor culture by Bacillus subtilis BK-17", J. Microbiol. Biotechnol., 9, pp.443~449 (1999)
  16. Lee, S. -K, "Purification and characterization of a fibrinolytic enzyme from Bacillus sp. KDO-13 isolated from soybean paste", J. Microbiol. Biotechnol., 11, pp.845~852 (2001)
  17. Liu, C. H, "Improvement in the growth performance of white shrimp, Litopenaeus vannamei, by a protease-producing probiotic,Bacillus subtilis E20, from natto", J. Appl. Microbiol., 107, pp.1031~1041 (2009)
  18. Liu, S, "Optimization of the production of organic solvent-stable protease by Bacillus sphaericus DS11 with response surface methodology", Bioresour. Technol., 101, pp.7924~7929 (2010)
  19. Nilegaonkar, S. S, "Production and partial characterization of dehairing protease from Bacillus cereus MCM B-326", Bioresour. Technol., 98, pp.1238~1245 (2007)
  20. Oskouie, S. F. G, "Response surface optimization of medium composition for alkaline protease production by Bacillus clausii", Biochem. Eng. J., 39, pp.37~42 (2008)
  21. PAIK, HYUN-DONG, "Purification and Characterization of the FibrinolyticEnzyme Produced by Bacillus subtilis KCK-7 from Chungkookjang", Journal of Microbiology and Biotechnology, 14(4), pp.829~835 (2004), 한국미생물·생명공학회
  22. Peng Y, "Purification and characterization of a fibrinolytic enzyme produced by Bacillus amyloliquefaciens DC-4 screened from douchi, a traditional Chinese soybean food", Compar. Biochem. Physiol., 134, pp.45~52 (2003)
  23. Veltman, O. R, "A single calcium binding site is crucial for the calcium-dependent thermal stability of thermolysin-like proteases", Biochem., 37, pp.5312~5319 (1998)
  24. 윤기홍, "식충식물로부터 Protease를 생산하는 Bacillus sp. SH-8의 분리와 효소 생산성", 한국미생물·생명공학회지, 34(4), pp.323~328 (2006), 한국미생물·생명공학회
  25. 윤경현, "한국재래간장에서 분리한 Bacillus amyloliquefaciens K42가 생산하는 혈전용해효소의 정제 및 특성", 한국미생물·생명공학회지, 31(3), pp.284~291 (2003), 한국미생물·생명공학회

관련 논문AI추천

이 간행물의 다른 논문 (19건)