DOI QR코드

DOI QR Code

Changes in Yeast and Bacterial Flora during Fermentation and Storage of Gugija-Liriope tuber Makgeolli using PCR-DGGE

PCR-DGGE를 이용한 구기자-맥문동 막걸리의 발효 과정과 저장 기간 중 효모와 세균 균총의 변화

  • Min, Jin-Hong (Department of Life Science and Genetic Engineering, Paichai University) ;
  • Nam, Yun-Gyu (Cheongyang Boxhorn Experimental Station, Chungnam Agriculture Research & Extension Service) ;
  • Ju, Jung-Il (Cheongyang Boxhorn Experimental Station, Chungnam Agriculture Research & Extension Service) ;
  • Jung, Jae-Hong (Cheongyang Green Wellness RIS, Cheongyang Provincial College) ;
  • Lee, Jong-Soo (Department of Life Science and Genetic Engineering, Paichai University) ;
  • Kim, Ha-Kun (Department of Life Science and Genetic Engineering, Paichai University)
  • 민진홍 (배재대학교 생명유전공학과) ;
  • 남윤규 (충남 농업기술원 청양 구기자 시험장) ;
  • 주정일 (충남 농업기술원 청양 구기자 시험장) ;
  • 정재홍 (청양대학 청양그린웰니스 RIS사업단) ;
  • 이종수 (배재대학교 생명유전공학과) ;
  • 김하근 (배재대학교 생명유전공학과)
  • Received : 2012.02.16
  • Accepted : 2012.03.26
  • Published : 2012.06.28

Abstract

In this study, we investigated the microbial flora changes in Gugija-Liriope tuber Makgeolli during fermentation and storage periods. We brewed Gugija-Liriope tuber Makgeolli for a week through twostage fermentations and stored the fermentation broth for a month at $4^{\circ}C$ or $20^{\circ}C$. We collected the samples periodically and analyzed microbial flora changes using viable cell counts and PCR-denaturing gradient gel electrophoresis (DGGE). Yeast viable cells were seen to have decreased to 13% of pre-storage levels after storage for 15 days at $20^{\circ}C$; however significant changes were not observed during storage at $4^{\circ}C$. Prolongation of storage time dramatically decreased the availability of viable cells. Yeast viable cell numbers had decreased to 38% of pre-storage levels at $4^{\circ}C$ and 4.8% at $20^{\circ}C$ after storage for 30 days. The results of the DGGE profile for yeast showed that Saccharomyces cerevisiae and Saccharomyces sp. were the predominant strains at the beginning of fermentation and throughout the whole period of storage. Viable cell counts for total bacteria had decreased to 36% of pre-storage levels after storage for 15 days but did not significantly change for the full 30 days of storage at $4^{\circ}C$. Similarly, viable cell counts for bacteria had decreased to 5% while viable cell numbers did not significantly change for the full 30 days at $20^{\circ}C$. Viable cell counts for lactic acid bacteria were performed and the results were similar to those for total bacteria. The results of the DGGE profile for bacteria showed that Weissella cibaria was the predominant strain at the beginning of fermentation. However it had disappeared by the end of fermentation, and Lactobacillus fermentum and Pediococcus acidilactici became the predominant species during storage.

구기자-맥문동 막걸리의 미생물 분포와 최적 저장 기간을 확립하기 위하여 먼저 구기자-맥문동 막걸리를 제조한 후 $4^{\circ}C$$20^{\circ}C$에 저장하면서 효모, 총 세균 그리고 젖산균의 생균 수 변화를 조사하였다. 발효가 완료되어 $4^{\circ}C$에서 15일간 저장하는 동안 효모 수는 큰 변화가 없었다. 하지만 30일간 저장한 후에는 약 62%가 감소되었다. $20^{\circ}C$에서 수행한 저장 실험에서는 15일간 저장한 후 초기 효모균수의 87%가 감소하였으며 30일간 저장한 후에는 약 95.2%가 감소되었다. 구기자-맥문동 막걸리의 DGGE 분석에 의해 S. cerevisiae와 미동정의 Saccharomyces sp.가 동정되었는데 $20^{\circ}C$에서 저장하였을 때Saccharomyces sp.가 더 빨리 사멸하였다. 저장 기간 중 총 세균 수와 젖산균의 변화 양상은 유사하였다. $4^{\circ}C$에서 저장 하였을 때 저장 15일 시점에서 총 세균 수는 약 64%가 감소하였으나 그 이후 저장 30일까지 총 세균수의 변화는 크게 없었다. $20^{\circ}C$에서 저장하였을 경우에는 초기 세균 수가 저장 15일 시점에서 총 세균수의 약 95%가 감소하였으며 그 이후 저장 30일 시점까지 총 세균수는 크게 변화하지 않았다. DGGE분석을 통해 발효 개시 시점에서는 W. cibaria가 우점균이었다가 점차 L. fermentum과 P. acidilactici가 증가하여 저장 기간 동안 이들이 우점균으로 존재 함을 확인하였다.

Keywords

References

  1. Baek, S. Y., Y. G. Nam, J. I. Ju, and J. S. Lee. 2011. Changes of quality characteristic during storage of Gugija-Liriope tuber Makgeolli made by Saccharomyces cerevisiae C-2. Kor. J. Mycol. 39: 122-125. https://doi.org/10.4489/KJM.2010.39.2.122
  2. Daeschel, M. A., and T. R. Klaenhaemmer. 1985. Association of a 13.6-megadalton plasmid in Pediococcus pentosaceus with bacteriocin activity. Appl. Environ. Microbiol. 50: 1538S-1541S
  3. Haruta, S., S. Ueno, I. Egawa, K. Hashiguchi, A. Fujii, M. Nagano, M. Ishii, and Y. Igarashi. 2006. Succession of bacterial and fungal communities during a traditional pot fermentation of rice vinegar assessed by PCR-mediated denaturing gradient gel electrophoresis. Int. J. Food Microbiol. 109: 79-87. https://doi.org/10.1016/j.ijfoodmicro.2006.01.015
  4. Lee, J. S., Y. C. Park, S. W. Paik, S. S. Lee, Y. K. Ahn, and J. S. Lee. 2008. Physiological functionality of Gugija products and an in vivo examination on anti-hypertension effects. J. Korean Soc. Food Sci. Nutr. 21: 115-120.
  5. Lee, M. Y., S. Y. Sung, H. K, Kang, H. S. Byun, S. M. Jung, J. H. Song, and J. S. Lee. 2010. Quality characteristics and physiological functionality of traditional rice wines in Chungnam province of Korean. Kor. J. Microbiol. Biotechnol. 38: 177-182.
  6. Lee, K. W., J. Y. Park, H. R. Jeong, H. J. Heo, N. S. Han, and J. H. Kim. 2012. Probiotic properties of Weissella strains isolated from human faeces. Anaerobe. 18: 96-102. https://doi.org/10.1016/j.anaerobe.2011.12.015
  7. Min, J. H., S. Y. Baek, J. S. Lee, and H. K. Kim. 2011. Changes of yeasts and bacterial flora during the storage of Korean traditional Makgeolli. Kor. J. Mycol. 39: 151-153. https://doi.org/10.4489/KJM.2010.39.2.151
  8. Muyzer, G., E. C. de Waal, and A. G. Uitterlinden. 1993. Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Appl. Environ. Microbiol. 59: 695-700.
  9. Park, W. J., B. C. Lee, J. C. Lee, E. N. Lee, J. E. Song, D. H. Lee, and J. S. Lee. 2007. Cardiovascular biofunctional activity and antioxidant activity of Gugija (Lycium chinensis Miller) species its hybrids. Kor. J. Medicinal. 15: 391-397.
  10. Reque, E. de F., A. Pandey, S. G. Franco, and C. R. Soccol. 2000. Isolation, identification and physiological study of Lactobacillus fermentum LPB for use as probiotic in chickens. Brazilian J. Microbiol. 31: 303-307.
  11. Rhee, I. J., and J. Y. An. 2003. Hepatoprotectives effects of water extract of Liriopis tuber on carbon tetrachlorideinduced hepatotoxicity in rats. Kor. J. Pharmacon. 34: 166-171.
  12. Song, J. H., J. H. Jang, H. K. Kim, and J. S. Lee. 2009. Manufacture and quality characteristic of Korean traditional big blue Lily-turf Takju. J. Natural Sciences. Paichai University, Korea. 20: 22-29.
  13. Song, J. H., S. Y. Baek, D. H. Lee, J. H. Jung, H. K. Kim, and J. S. Lee. 2011. Screening of fungal Nuruk and yeast for brewing of Gugija-Liriope tuber traditional rice wine and optimal fermentation condition. Kor. J. Mycol. 39: 78-84 https://doi.org/10.4489/KJM.2011.39.1.078

Cited by

  1. 시판 막걸리의 이화학적 특성과 젖산균 함량 및 생리기능성 vol.40, pp.4, 2012, https://doi.org/10.4014/kjmb.1207.07006
  2. 전통발효식품에서 분리한 진균류와 세균을 이용하여 제조한 누룩과 막걸리 그리고 청국장의 생리기능성 vol.40, pp.3, 2012, https://doi.org/10.4489/kjm.2012.40.3.164
  3. 구기자 잎 막걸리 제조용 효모의 선발 및 최적 알코올 발효조건 vol.41, pp.3, 2013, https://doi.org/10.4489/kjm.2013.41.3.167
  4. 맥문동의 물추출액 및 에탄올추출액 이용 음료의 산화방지 활성 vol.30, pp.6, 2012, https://doi.org/10.9724/kfcs.2014.30.6.785
  5. Characterization of a minimal pKW2124 replicon from Weissella cibaria KLC140 and its application for the construction of the Weissella expression vector pKUCm1 vol.6, pp.None, 2012, https://doi.org/10.3389/fmicb.2015.00035
  6. 알코올 농도와 저장온도가 탁주 품질에 미치는 영향 vol.44, pp.10, 2012, https://doi.org/10.3746/jkfn.2015.44.10.1531
  7. Antibacterial and antioxidative activities of licorice extracts fermented with Nuruk molds vol.25, pp.7, 2012, https://doi.org/10.11002/kjfp.2018.25.7.830
  8. Quality characteristics and antioxidant activities of aged black Liriope platyphylla vol.26, pp.5, 2012, https://doi.org/10.11002/kjfp.2019.26.5.505