Browse > Article
http://dx.doi.org/10.5352/JLS.2012.22.1.110

Analysis of Microbial Diversity in Nuruk Using PCR-DGGE  

Kwon, Seung-Jik (Division of Bacterial Respiratory Infections, Korea National Institute of Health)
Sohn, Jae-Hak (Department of Bio-food material, College of Medical & Life Sciences, Silla University)
Publication Information
Journal of Life Science / v.22, no.1, 2012 , pp. 110-116 More about this Journal
Abstract
Nuruk plays a significant role in the flavor and quality of Takju and Yakju, which are produced through saccharification and alcohol fermentation by various microorganisms. In this study, we identified microbial strains isolated from a plate count and PCR-denaturing gradient gel electrophoresis (DGGE) analysis targeting the 16S and 28S rRNA genes, in order to characterize bacterial and fungal diversity in Sansung Nuruk. The numbers of bacteria and fungi in Nuruk were $1.5{\times}10^9$ CFU/g and $2.2{\tims}10^8$ CFU/g, respectively. The 16S rRNA gene sequence indicated that the predominant bacteria in the isolates and PCR-DGGE profile of Nuruk were Kocuria spp., Pantoea spp., Lactobacillus spp., Pediococcus spp., Weissella spp., Staphylococcus spp., endophytic bacterium, uncultured Gamma-proteobacteria, uncultured Cyanobacteria, and Actinobacteria. Dominant bacteria from the PCR-DGGE profile were Pediococcous pentosaceus and uncultured Cyanobacteria. The 28S rRNA gene sequence indicated the predominant fungi in the isolates and PCR-DGGE profile to be Trichomonascus spp. Pichia spp., Torulaspora spp., Wickerhamomyces spp., Sacharomycopsis spp., Lichtheimia spp., Mucor spp., Rhizopus spp. Aspergillus spp., and Cladosporium spp. Dominant fungi from the PCR-DGGE profile were Pichia kudriavzevii and Aspergillus oryzae. The PCR-DGGE technique was used for the first time in this study to assess a microbial community in Nuruk and proved to be an effective protocol for profiling microbial diversity.
Keywords
Denaturing Gradient Gel Electrophoresis (DGGE); Nuruk; 16S rRNA gene; 28S rRNA gene; Takju;
Citations & Related Records
Times Cited By KSCI : 11  (Citation Analysis)
연도 인용수 순위
1 Meroth, C. B., W. P. Hammes, and C. Hertel. 2003. Identification and population dynamics of yeasts in sourdough fermentation processes by PCR-denaturing gradient gel electrophoresis. Appl. Environ. Microbiol. 69, 7453-7461.   DOI   ScienceOn
2 Mills, D. A., E. A. Johannsen, and L. Cocolin. 2002. Yeast diversity and persistence in botrytis-affected wine fermentations. Appl. Environ. Microbiol. 68, 4884-4893.   DOI   ScienceOn
3 Muyzer, G., E. D. 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.
4 Rantsiou, K., R. Urso, L. Iacumin, C. Cantoni, P. Cattaneo, G. Comi, and L. Cocolin, 2005. Culture-dependent and -independent methods to investigate the microbial ecology of Italian fermented sausages. Appl. Environ. Microbiol. 71, 1977-1986.   DOI   ScienceOn
5 Reeson, A. F., T. Jankovic, M. L. Kasper, S. Rogers, and A. D. Austin. 2003. Application of 16S rDNA-DGGE to examine the microbial ecology associated with a socialwasp Vespula germanica. Insect Mol. Biol. 12, 85-91.   DOI   ScienceOn
6 Lee, S. H., H. J. Jung, S. H. Yeo, H. S. Kim, and T. S. Yu. 2004. Characteristics of a-amylase of a new species Aspergillus coreanus NR 15-1. Korean J. Biotechnol. Bioeng. 19, 301-307.   과학기술학회마을
7 Kim, H. S., J. S. Hyun, J. Kim, H. P. Ha, and T. S. Yu. 1998. Enzymological characteristics and identification of useful fungi isolated from traditional Korean nuruk. Korean J. Appl. Microbiol. Biotechnol. 26, 456-464.   과학기술학회마을
8 Kim, J. Y. and J. S. Kho. 2004. Screening of brewing yeasts and saccharifying molds for foxtail millet-wine making. J. Korean Soc. Appl. Biol. Chem. 47, 78-84.   과학기술학회마을
9 Marmur, J. 1961. A procedure for the isolation of DNA from microorganisms. J. Mol. Biol. 3, 208-218.   DOI
10 Masoud, W., L. B. Cesar, L. Jespersen, and M. Jakobsen. 2004. Yeast involved in fermentation of Coffea arabica in East Africa determined by genotyping and by direct denaturating gradient gel electrophoresis. Yeast 21, 549-556.   DOI   ScienceOn
11 Yang, J. Y. and K. H. Lee. 1996. Shelf-life and microbiological study of Sansung Takju. Korean J. Food Sci. Technol. 28, 779-785.   과학기술학회마을
12 Bowman, J. P., S. A. McCammon, J. A. Gibson, L. Robertson, and P. D. Nichols. 2003. Prokaryotic metabolic activity and community structure in Antarctic continental shelf sediments. Appl. Environ. Microbiol. 69, 2448-2462.   DOI   ScienceOn
13 Chang, H. W., K. H. Kim, Y. D. Nam, S. W. Roh, M. S. Kim, C. O. Jeon, H. M. Oh, and J. W. Bae. 2008. Analysis of yeast and archaeal population dynamics in kimchi using denaturing gradient gel electrophoresis. Int. J. Food Microbiol. 126, 159-166.   DOI   ScienceOn
14 Cocolin, L., L. F. Bisson, and D. A. Mills. 2000. Direct profiling of the yeast dynamics in wine fermentations. FEMS Microbiol. Lett. 189, 81-87.   DOI   ScienceOn
15 Thompson, J. D., T. J. Gibson, F. Plewniak, F. Jeanmougin, and D. G. Higgins. 1997. The CLUSTAL_$\chi$ windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. 25, 4876-4882.   DOI   ScienceOn
16 Xia, S., Y. Shi, Y. Fu, and X. Ma. 2005. DGGE analysis of 16S rDNA of ammonia-oxidizing bacteria in chemical-biological flocculation and chemical coagulation systems. Appl. Environ. Microbiol. 69, 99-105.
17 Yeates, C., M. R. Gillings, A. D. Davison, N. Altavilla, and D. A. Veal. 1998. Methods for microbial DNA extraction from soil for PCR amplification. Biol. Proced. Online 1, 40-47.   DOI   ScienceOn
18 Yu, T. S., H. S. Kim, H. P. Ha, T. Y. Kim, and I. W. Yoon. 1996. Bibliographical study on microorganisms of Nuruk (Until 1945). J. Korean Soc. Food Nutr. 25, 170-179.   과학기술학회마을
19 Shin, Y. D. and D. H. Cho. 1970. A study on the microflora changes during Takju brewing. Korean J. Microbiol. 8, 53-64.   과학기술학회마을
20 Sharma, S., Z. Szele, R. Schilling, J. C. Munch, and M. Schloter. 2006. Influence of freeze-thaw stress on the structure and function of microbial communities and denitrifying populations in soil. Appl. Environ. Microbiol. 72, 2148-2154.   DOI   ScienceOn
21 So, M. H. and Y. S. Lee. 2009. Effects of culture conditions of Rhizopus sp. ZB9 on the production of saccharifying amylase during the preparation of rice koji. Korean J. Food Nutr. 22, 644-649.   과학기술학회마을
22 Suh, M. J. and K. H. Nho. 1987. Studies on the screening and properties of raw starch saccharifying microorganism. Korean J. Mycol. 15, 169-174.   과학기술학회마을
23 Ha, D. M., D. C. Kim, S. M. Hong, and C. W. Lee. 1989. Identification and properties of starch utilizing yeasts isolated from Nuruk. J. Korean Agric. Chem. Soc. 32, 408-415.   과학기술학회마을
24 Jo, K. Y. and D. M. Ha. 1995. Isolation and identification of the Lactic acid bacteria from Nuruk. Agri. Chem. Biotechnol. 38, 95-99.   과학기술학회마을
25 Jung, H. K., C. H. Park, G. D. Lee, S. C. Park, H. H. Park, and J. H. Hong. 2007. Characteristics of Pichia anomala K15 producing killer toxin isolated from traditional nuruk. J. Korean Soc. Food Sci. Nutr. 36, 1077-1082.   과학기술학회마을   DOI
26 Lane, D. J. 1991. 16S/23S rRNA sequencing, pp. 115-175, In Stackebrandt, E. and M. Goodfellow (eds.), Nucleic acid techniques in bacterial systematic, John Wiley & Sons. New York.