Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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Chemical changes in resazurin by probiotics and its application for evaluating living bacterial cell counts and their reduction potentials

프로바이오틱스에 의한 레자주린의 화학적 변화와 생균수 및 환원활성 측정에의 적용

  • Lee, Hyowon (Division of Applied Food System, College of Natural Science, Seoul Women's University) ;
  • Oh, Yeong Ji (Department of Food and Animal Biotechnology, Center for Food and Bioconvergence, Seoul National University) ;
  • Hong, Jungil (Division of Applied Food System, College of Natural Science, Seoul Women's University)
  • 이효원 (서울여자대학교 자연과학대학 식품응용시스템학부) ;
  • 오영지 (서울대학교 식품바이오융합연구소) ;
  • 홍정일 (서울여자대학교 자연과학대학 식품응용시스템학부)
  • Received : 2021.01.12
  • Accepted : 2021.02.17
  • Published : 2021.04.30
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Abstract

Resazurin, an oxidized blue dye, is reduced to resorufin, showing a peak absorbance change and emitting fluorescence due to the metabolic activity of living cells. In this study, the growth of Lactobacillus rhamnosus GG (LGG) and the redox potential of living probiotic bacteria were evaluated based on changes in the absorbance or fluorescence of resazurin. Fluorescence analysis is a more accurate and sensitive method for quantifying viable LGG than the colorimetric absorbance measurement of resazurin change. Fluorescence measurement could detect LGG of 6.5-9.5 log CFU/mL within 30 min with R2=0.99. No significant effect of further reduction of resorufin to dihydroresorufin by LGG was observed. Various probiotics showed different resazurin-reducing activities, and L. kimchicus had the highest reducing activity among the six probiotics tested. These findings suggests that fluorescence measurement in a resazurin-based assay is useful for analyzing bacterial growth and the redox potential of living probiotics.

본 연구에서는 살아있는 프로바이오틱스 균주에 의한 레자주린의 흡광 및 형광특성의 변화와 레소루핀과의 반응성을 분석하고, 균주별 레자주린에 대한 환원능을 비교하였다. LGG에 의해 레자주린은 흡광과 형광의 변화를 수반하며 레소루핀으로 환원되고 반응시간과 생균수의 증가에 따라 환원정도가 증가하였으며, 형광의 변화에서 더 정확하고 민감한 반응성을 보였다. 한편 LGG에 의한 레소루핀으로부터 다이하이드로레소루핀으로의 환원반응은 거의 유발되지 않았다. 프로바이오틱스 6개 균주 중 L. kimchicus의 생균에 의한 레자주린 환원력이 월등하게 높은 반면, 균체 파쇄 후의 ABTS 및 DPPH 라디칼 소거능은 다른 양상을 보이며 L. plantarum과 L. casei가 높은 활성을 나타냈다. 한편 생균의 MTT 환원능은 L. kimchicus가 LGG에 비해 현저히 높아 레자주린 환원능과 유사한 양상을 보였다. 본 연구결과는 레자주린이 단일 균주 프로바이오틱스의 생균수 측정에 유용하며, 프로바이오틱스의 선별 및 환원활성 측정에 활용될 수 있음을 시사한다.

Keywords

References

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