Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Isolation of Bacteria from Chunggukjang Prepared by Rice Straw and Identification of Protease Secreted

볏짚 청국장 발효 세균 분리 및 분비된 protease의 확인

  • Oh, Jae-Hyeon (Department of Biological Sciences, Sunchon National University) ;
  • Lee, Byeong-Jeong (Department of Biological Sciences, Sunchon National University) ;
  • Paik, Hyoung-Rok (Department of Biological Sciences, Sunchon National University) ;
  • Jung, Sang-Chul (Department of Biological Sciences, Sunchon National University) ;
  • Baik, Keun-Sik (Department of Biological Sciences, Sunchon National University) ;
  • Choi, Sang-Ki (Department of Biological Sciences, Sunchon National University)
  • Published : 2009.03.31
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Abstract

To isolate bacteria secreting protease, which can dissolve fibrin efficiently, we prepared chunggukjang using rice straw and isolated, preliminarily, approximately 100 bacterial stains. Their capabilities to dissolve milk protein as well as fibrin included in media were then examined and finally, five strains named J1 - J5 were selected. Among them, J-4, which is close to bacillus subtilis, showed highest activity for fibrin dissolution. Proteases secreted from the J-4 strain were partially purified from culture supernatant using DEAE-sepharose column chromatography and identified with SDS-polyacrylamide gel electrophoresis. Three proteins were subjected to analysis with MALDI-TOF and PMF (Peptide Mass Fingerprinting). 41.9 kDa protein was identified as a neutral protease. On the other hand, 45 kDa protein turned out to be bacillopeptidase F, with a molecular mass of 91.7 kDa, indicating that partially purified peptide is a degradation product.

청국장에서 혈전 용해능이 우수한 균주를 분리하기 위해 짚을 이용하여 직접 청국장을 제조하였으며 이로부터 1, 000여종의 균주를 1차적으로 분리하였다. 2차적으로 skim milk가 첨가된 배지 및 fibrin 배지의 단백질분해 실험을 통해 혈전 용해능이 우수한 균주를 분리하였다. 이 과정을 통해 선발된 균주는 J-1, J-2, J-3, J-4, J-5로 명명된 5종 균주이었으며 그 중 Bacillus 계통의 J-4 균주가 가장 활성이 높은 균주로 선별되었다. Bacillus subtilis J-4 균주가 생산하는 protease를 DEAE-sepharose column을 사용하여 부분 정제 분리한 결과 SDS-PAGE Gel 상에서 45.0 kDa의 질량이었다. 이 단백질을 MALDI-TOF 및 PMF(Peptide Mass Fingerprinting)를 사용하여 분석한 결과 neutral protease와 bacillopeptidase F가 확인되었다.

Keywords

References

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