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말로락틱 발효에 적합한 토착 Lactobacillus plantarum 분리

Isolation of indigenous Lactobacillus plantarum for malolactic fermentation

  • 허준 (전북대학교 자연과학대학 생명과학과) ;
  • 이찬미 (전북대학교 자연과학대학 생명과학과) ;
  • 박문국 (전북대학교 자연과학대학 생명과학과) ;
  • 정도연 ((재)발효미생물산업진흥원) ;
  • 엄태붕 (전북대학교 자연과학대학 생명과학과)
  • Heo, Jun (Department of Biological Sciences, Chonbuk National University) ;
  • Lee, Chan-Mi (Department of Biological Sciences, Chonbuk National University) ;
  • Park, Moon Kook (Department of Biological Sciences, Chonbuk National University) ;
  • Jeong, Do-Youn (Microbial Institute for Fermentation Industry (MIFI)) ;
  • Uhm, Tai-Boong (Department of Biological Sciences, Chonbuk National University)
  • 투고 : 2015.06.02
  • 심사 : 2015.06.19
  • 발행 : 2015.06.30

초록

말로락틱 발효(MLF)은 유산균의 말로락틱 효소(Mle)에 의해 malic acid가 lactic acid로 전환되는 과정으로 와인 제조에 널리 사용된다. 전통 발효 식품으로부터 54개의 유산균을 분리한 다음 MLF 특성을 가진 균주를 선발하기 위해 Lactobacillus plantarum mle 유전자 서열의 보존 영역에 대한 primer 쌍을 제작했고, PCR을 통해 이 유전자를 함유한 4 종의 균주를 선발하였다. 선발된 균주들의 16S rRNA 염기서열과 생화학적 특성, rec gene 영역의 PCR을 수행하여 동정한 결과 Lactobacillus plantarum으로 모두 동정되었다. 1,644 bp로 구성된 이들 mle 유전자의 분석 결과 JBE60 균주의 염기 서열은 JBE150, JBE160, JBE171 균주들과 96.7%, 아미노산 서열로는 99.5%가 일치했다. 에탄올 저항성을 확인한 결과 JBE60 균주가 10% 에탄올에 대한 저항성이 가장 높았다. MLF 활성을 확인한 결과 이들 균주는 평균 43%의 malic acid 감소를 보였으며 균주 간 분해율은 비슷했다. 이러한 결과로부터 JBE60 균주가 와인용 MLF 종균으로 이용 될 수 있을 것으로 보인다.

The malolactic fermentation (MLF), which is widely used in winemaking, is the conversion of malic acid to lactic acid conducted by the malolactic enzyme (Mle) of lactic acid bacteria. In order to select the strains with MLF among 54 lactic acid bacteria isolated from the traditionally fermented foods, we designed a primer set that specifically targets the conserved regions of the mle gene and then selected four strains that harbor the mle gene of Lactobacillus plantarum. All strains were identified as L. plantarum by analyzing the 16S rRNA sequences, biochemical properties, and the PCR products of the recA gene. From comparison of the mle gene sequences consisting of 1,644 bp, the nucleotide and amino acid sequence of strain JBE60 correspond to 96.7% and 99.5% with those of other three strains, respectively. The strain JBE60 showed the highest resistant against 10% (v/v) ethanol among the strains. The strains lowered the concentration of malic acid to average 43%. Considering the ethanol resistance and conversion of malic acid, the strain JBE60 is considered as a potential starter for the malolactic fermentation.

키워드

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