Journal of Animal Science and Technology

Korean Society of Animal Sciences and Technology (한국축산학회)
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Expression of Clostridium thermocellum Endoglucanase Gene in Lactobacillus bulgaricus and Lactobacillus plantarum and in vitro Survival Characteristics of the Transformed Lactobacilli

Lactobacillus bulgaricus와 Lactobacillus plantarum 균주에서 Clostridium thermocellum 유래 endoglucanase의 발현과 발현 유산균의 in vitro 생존 특성

  • Cho, J.S. (School of agricultural biotechnology, Seoul National University) ;
  • Kang, S.H. (School of agricultural biotechnology, Seoul National University) ;
  • Lee, H.G. (School of agricultural biotechnology, Seoul National University) ;
  • Lee, H.J. (National Livestock Research Institute, RDA) ;
  • Woo, J.H. (Laboratory of Molecular Biology, National Institute of Mental Health) ;
  • Moon, Y.S. ;
  • Yang, C.J. (Department of Animal Science and Biotechnology, Jinju National University) ;
  • Choi, Y.J. (Department of Animal Science and Technology, Sunchon National University)
  • 조재순 (서울대학교 농생명공학부) ;
  • 강승하 (서울대학교 농생명공학부) ;
  • 이홍구 (서울대학교 농생명공학부) ;
  • 이현준 (농촌진흥청 축산기술연구소) ;
  • 우정희 (미국 메릴렌드주 국립보건원) ;
  • 문양수 (진주산업대학교 동물생명과학과) ;
  • 양철주 (순천대학교 동물자원과학과) ;
  • 최윤재 (서울대학교 농생명공학부)
  • Published : 2003.08.30
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Abstract

Endoglucanase A from Clostridium thermocellum which is resistant to pancreatic proteinase was selected out of numbers cellulases then were expressed in lactobacilli. Recombinant lactobacilli expression vector, pSD1, harboring the endoglucanase gene from C. thermocellum under the control of its own promoter, was constructed. Both L. bulgaricus and L. plantarum were electrotransformed with pSD1. The endoglucanase activities of 0.120 and 0.144 U/ml were found in culture media of L. bulgaricus and L. plantarum containing pSD1, respectively. In vitro survival characteristics of the transformed lactobacilli were tested. Both L. bulgaricus and L. plantarum showed a similar resistance to low pH 3. Moreover, L. plantarum was bile-salt resistant in the presence of 0.3 and 1% oxgall. L. bulgaricus and L. plantarum showed a rather homogenous resistant pattern against the tested antibiotics. Both of the strains were resistant to amikacin, gentamicin, streptomycin, kanamycin, and colistin.

다양한 미생물들로부터 유래한 cellulase 중에서, 특히 장내 단백질 가수분해효소에 안정한 Clostridium thermocellum 균주 유래의 endoglucanase를 선별하였다. 그 후 그 유전자의 자체 프로모터에 의해 발현되는 재조합 Lactobacillus용 발현벡터를 구축하였고, 그 발현벡터를 pSD1이라 명명하였다. 이 발현벡터를 L. bulgaricus와 L. plantarum 균주에 각각 전기천공법을 이용하여 형질전환시키는데 성공하였으며 그 재조합 균주들로부터 endoglucanase 효소역가를 조사한 결과 각각 배지 상층액에서 0.12, 0.144 U/ml로 조사되었다. 한편 이들 균주들의 생균제로 갖추어야할 특성인 내산성, 내담즙성 및 항생제내성 여부를 조사한 결과, 이들 균주들은 모두 pH3과 같은 산성 조건하에서도 안정하였으며, 내담즙성에 있어서는 특히 L. plantarum 균주의 경우 0.3, 1% 의 oxgall에서도 안정하였다. 또한 항생제 내성을 조사한 결과 두 균주 모두 amikacin, gentamicin, kanamycin, colistin에 저항성이 높은 것으로 나타났다.

Keywords

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