Fibrinolytic, Immunostimulating, and Cytotoxic Activities of Microbial Strains Isolated from Kochujang

고추장 분리 균주의 혈전용해능, 면역활성능 및 세포독성 효과 조사

  • Published : 2007.06.30

Abstract

This study was carried out to investigate the functional activities of microorganisms isolated from kochujang, such as fibrinolytic, immunostimulating, and cytotoxical activities, and to apply these microorganisms to kochujang products. Ninety-one microbial strains with proteolytic activity were selected from 294 strains isolated from traditional and commercial kochujang. Three strains (TPP 0014, TPP 6013, and TPP 6015) with high fibrinolytic activity were tested for their immunostimulating and cytotoxical activities. For the assessment of macrophage activation, cytokines such as tumor necrosis factor, $interleukin-1{\alpha}$ and nitrogen oxide were measured with the murine macrophage cell line RAW 264.7. In addition, the cytotoxical activities of the three strains were examined by MTT assay on the colon cancer cell line SNU-C4 and normal cell line CHO-K1. Using an API identifying kit, two of the microbial strains (TPP 0014 and TPP 6015) were identified as Bacillus stearothermophilus and the other strain (TPP 6013) was identified as B. amyloliquefacience.

본 연구는 고추장에서 분리한 미생물들을 이용하여 혈전용해능, 면역세포활성능 및 세포독성과 같은 건강기능활성을 조사하고, 건강기능활성 미생물을 고추장에 응용하고자 수행되었다. 전통식 및 개량식 고추장으로부터 분리된 294균주 중 단백질분해력이 우수한 91균주를 선별하였고, 선별된 91개 균주의 혈전용해능을 측정하여 최종적으로 혈전용해능 활성이 높은 3 균주(TPP 0014, TPP 6013, TPP 6015)를 선별하였다. 이들 최종 선별균주들의 면역세포활성화능력 측정 결과 TPP 6015 균주에 의한 거식세포에 의한 NO 생성량은 3.67 ${\mu}g/mL$이었고, $TNF-{\alpha}$ 생성량은 626.3 pg/mL 그리고 $IL-{\alpha}$ 생성량은 645.9 pg/mL로 측정되어 세 균 주 중 비교적 높은 면역세포활성이 있음을 확인할 수 있었다. 또한 대장암 세포주인 SNU-C4와 정상세포주인 CHO-K1에 대한 선별된 균주의 세포독성을 MTT assay에 의해 조사한 결과 SNU-C4 세포주에 65-78%, 그리고 CHO-K1 세포주에 60-70%의 세포독성을 나타내는 것을 확인하였다. 고추장 시료는 혈전용해 활성을 나타내지 않았으나, 이들 선발 균주들을 고추장에 첨가하였을 때, 높은 혈전용해활성을 나타내는 것으로 밝혀졌다. 이들 선별 균주들을 API kit를 이용하여 동정한 결과 TPP 0014와 TPP 6015 균주는 Bacillus stearothermophilus로 동정이 되었고, TPP 6013 균주는 B. amyloliquefacience로 동정이 되었다.

Keywords

References

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