Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Isolation and Characterization of Exopolysaccharide Producing Lactic Acid Bacteria from Kimchi

김치로부터 Exopolysaccharide 생성 유산균의 분리 및 특성 규명

  • Kim Uyo-Ju (Department of Food and Nutrition, Chosun University) ;
  • Chang Hae-Choon (Department of Food and Nutrition, Chosun University)
  • 김효주 (조선대학교 식품영양학과) ;
  • 장해춘 (조선대학교 식품영양학과)
  • Published : 2006.09.01
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Abstract

Three slime-forming lactic acid bacteria were isolated from Kimchi and shown to produce viscous exopolysaccharides (EPS) in sucrose media. The isolated strains, GJ2, C3 and C11, were identified as Leuconostoc kimchii, Leuconostoc citreum and Leuconostoc mesenteroides, respectively, by examining their metabolic characteristics and determining their 16S rDNA sequences. Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 exhibited high viability (maintained initial viable cell count of $10^8$ CFU/ml) in 0.05 M sodium phosphate buffer (pH 3.0) for 2 h, in artificial gastric juice for 2 h and in 0.3% oxgall for 24 h. When tested, Leu. kimchii GJ2, in particular, displayed antimicrobial activity against pathogenic microorganisms. Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 produced 21.49 g/l, 16.46 g/l and 22.98 g/l EPS, respectively, in sucrose (5%) medium. The amount of purified EPS extracted from Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 was 14.61 g/l, 7.73 g/l and 4.77 g/l, respectively. Although the EPS produced by Leu. kimchii GJ2, Leu. citreum C3 and Leu. mesenteroides C11 differed in viscosity, TLC and HPLC analysis revealed that each contained only one type of monosaccharide, glucose. The average molecular mass of EPS produced by Leu. kimchii GJ2 was 306,606 Da.

김치로부터 3종의 EPS 생성 유산균을 분리하였다. 분리균주는 Leuconostoc kimchii, Leuconostoc citreum, Leuconostoc mesenteroides로 동정되었으며, 각각 Leuconostoc kimchii GJ2, Leuconostoc citreum C3, Leuconostoc mesenteroides C11로 명명하였다. EPS생성 김치유산균의 장내 정착성 여부 확인 결과 pH 3.0의 0.05 M sodium phosphate buffer와 인공위액에서 2시간 처리한 후에도 3균주 모두 초기균수($10^8\;CFU/ml$)를 유지하였으며 인공담즙에서 24시간 처리한 후에도 3균주 모두 초기균수($10^8\;CFU/ml$)를 유지하며 3종의 EPS생성 김치유산균 Leu. kimchii GJ2, Leu. citreum C3, Leu. mesenteroides C11이 장내 정착하면서 probiotics로서 작용할 수 있을 것임을 확인하였다. 반면, 대조군으로서 분리균주가 EPS를 생성하지 않았을 때의 경우에는 내산성, 인공위액, 인공담즙에서 처리 후 생균수가 $10^{1-2}\;CFU/ml$ 감소현상을 보여 균체의 EPS생성 여부가 probiotics로서 기능할 수 있는데 결정적 요인임을 보여주었다. 특히 Leu. kimchii GJ2의 경우 E. coli, Pseudomonas, Listeria, Micrococcus, Salmonella, Staphylococcus, Streptococcus속 등의 유해 균주에서 항균활성을 나타내었으며, 생산 항균물질은 단백질성 물질로 확인되었다. 김치유산균 Leu. kimchii GJ2, Leu. citreum C3, Leu. mesenteroides C11로부터의 EPS 생성량은 sucrose(5%) 배지에서 각각 21.49 g/l, 16.46 g/l, 22.98 g/l 였으며, 정제 시에도 각각 14.61 g/l, 7.73 g/l, 4.77 g/l로 기존의 EPS 생성 유산균에서의 생산량에 비해 10배 이상의 높은 생산량을 나타내었다. TLC 및 HPLC를 이용한 EPS 구성 당 확인 결과 3균주 모두 glucose로만 구성된 homopolysaccharides로 확인되었다. EPS 생성량이 가장 좋은 Leu. kimchii GJ2의 평균 분자량은 360,606 Da이었으며, 나머지 두 균주에 대해서는 생성 EPS 형태와 점도의 차이로 미루어 보아 생성 EPS의 분자구조와 분자량이 서로 다른 것으로 판단하였다.

Keywords

References

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