생명과학회지 (Journal of Life Science)

  • 제22권2호
  • /
  • Pages.232-238
  • /
  • 2012
  • /
  • 1225-9918(pISSN)
  • /
  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
권호 표지
DOI QR코드

DOI QR Code

PCR-DGGE를 이용한 막걸리발효에서 미생물 다양성 분석

Analysis of Microbial Diversity in Makgeolli Fermentation Using PCR-DGGE

  • 권승직 (국립보건연구원 질병관리본부) ;
  • 안태영 (단국대학교 미생물학과) ;
  • 손재학 (신라대학교 의생명과학대 바이오식품소재학과)
  • Kwon, Seung-Jik (Division of Bacterial Respiratory Infections, Korea National Institute of Health) ;
  • Ahn, Tae-Young (Department of Microbiology, Dankook University) ;
  • Sohn, Jae-Hak (Department of Bio-food material, College of Medical & Life Sciences, Silla University)
  • 투고 : 2011.12.21
  • 심사 : 2012.02.23
  • 발행 : 2012.02.28
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

금정산성 막걸리$^{(R)}$는 전통적인 수제누룩과 쌀로부터 발효된 한국의 전통적인 술이다. 본 연구에서는 막걸리 발효기간 동안 세균과 진균의 다양성을 특성화하기 위해 16S와 28S rRNA 유전자를 목적으로 하는 PCRDenaturing Gradient Gel Electrophoresis (PCR-DGGE) 분석을 수행하였다. 막걸리 발효기간 동안 PCR-DGGE profile에서 검출된 세균은 16S rRNA 유전자 서열에 기초한 동정결과 Lactobacillus spp. (L. curvatus, L. kisonensis, L. plantarum, L. sakei 및 L. gasseri), Pediococcus spp. (P. acidilactici, P. parvulus, P. agglomerans및 P. pentosaceus), Pantoea spp. (P. agglomerans 및 P. ananatis) 그리고 Citrobacter freundii로 총 12종이었으며, 배양2일 이후 L. curvatus가 주된 우점 종을 형성하였다. 반면 PCR-DGGE profile에서 검출된 진균은 28S rRNA 유전자 서열에 기초한 동정결과 Pichia kudriavzevii, Saccharomyces cerevisiae, Asidia idahoensis, Kluyveromyces marxianus, Saccharomycopsis fibuligera 및 Torulaspora delbrueckii로 6종이었으며 주된 우점 진균은 배양0일에서 2일에 P. kudriavzevii에서 배양 3일에서 6일에 S. cerevisiae로 전환되었다. 결과적으로 PCR-DGGE분석은 막걸리발효기간 동안 미생물의 구조와 다양성을 이해하는 데 유용한 도구임을 보여주었다.

Kumjungsansung-Makgeolli$^{(R)}$ is a traditional Korean rice wine that is fermented from traditional nuruk and rice. In this study, we performed the PCR-denaturing gradient gel electrophoresis (DGGE) analysis targeting the 16S and 28S rRNA genes to characterize bacterial and fungal diversity during Makgeolli fermentation. The predominant bacteria in the PCR-DGGE profile during Makgeolli fermentation were Lactobacillus spp. (Lactobacillus curvatus, L. kisonensis, L. plantarum, L. sakei, and L. gasseri), Pediococcus spp. (P. acidilactici, P. parvulus, P. agglomerans, and P. pentosaceus), Pantoea spp. (P. agglomerans and P. ananatis), and Citrobacter freundii; these were identified on the base of analysis of 16S rRNA gene sequences. The dominant bacterium during Makgeolli fermentation was L. curvatus. The predominant fungi in PCR-DGGE profile during Makgeolli fermentation were Pichia kudriavzevii, Saccharomyces cerevisiae, Asidia idahoensis, Kluyveromyces marxianus, Saccharomycopsis fibuligera, and Torulaspora delbrueckii, and these were identified on the basis of analysis of 28S rRNA gene sequences. The dominant fungal species during Makgeolli fermentation changed from P. kudriavzevii at 0-2 days incubation to S. cerevisiae at 3-6 days incubation. This study suggests that PCR-DGGE analysis could be a suitable tool for the understanding of microbial diversity and structure during Makgeolli fermentation.

키워드

참고문헌

  1. Baek, S. Y., Y. G. Nam, J. I. Ju, and J. S. Lee. 2011. Changes of quality characteristics during storage of Gugija-Liriope tube Makgeolli made by Saccharomyces cerevisiae C-2. Korean J. Mycol. 39, 122-125. https://doi.org/10.4489/KJM.2010.39.2.122
  2. 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. https://doi.org/10.1128/AEM.69.5.2448-2462.2003
  3. 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. https://doi.org/10.1016/j.ijfoodmicro.2008.05.013
  4. Cho, E. K., H. Y. Kim, H. J. Byeon, S. W. Kim, and Y. J. Choi. 2010. Nitrite scavenging and alcohol metabolizing activities of hot water extract from Makgeoly and its angiotensin converting enzyme inhibitory effect. J. Life Sci. 20, 768-774. https://doi.org/10.5352/JLS.2010.20.5.768
  5. 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. https://doi.org/10.1111/j.1574-6968.2000.tb09210.x
  6. El-Baradei, G., A. Delacroix-Buchet, and O. Jean-Claude. 2007. Biodiversity of bacterial ecosystems in traditional Egyptian domiati cheese. Appl. Environ. Microbiol. 73, 1248-1255. https://doi.org/10.1128/AEM.01667-06
  7. Galvez, A., H. Abriouel, R. L. Lopez, and N. B. Omar. 2007. Bacteriocin-based strategies for food biopreservation. Int. J. Food Microbiol. 120, 51-70. https://doi.org/10.1016/j.ijfoodmicro.2007.06.001
  8. Heuer, H. and K. Smalla. 1997. Application of denaturing gradient gel electrophoresis (DGGE) and temperature gradient gel electrophoresis (TGGE) for studying soil microbial communities, pp. 353-373. In van Elsas, J. D., E. M. H. Wellington, and J. T. Trevors (eds.), Modern soil microbiology. Marcel Dekker, Inc., New York, N.Y.
  9. Jeong, J. W., K. J. Park, M. H. Kim, and D. S. Kim. 2006. Quality characteristics of Takju fermentation by addition of chestnut peel powder. Korean J. Food Preserv. 13, 329-336.
  10. Jin, J., S. Y. Kim, Q. Jin, H. J. Eom, and N. S. Han. 2008. Diversity analysis of lactic acid bacteria in Takju, Korean rice wine. J. Microbiol. Biotechnol. 18, 1678-1682.
  11. Jo, K. Y. and D. M. Ha. 1995. Isolation and identification of the Lactic acid bacteria from Nuruk. Agri. Chem. Biotechnol. 38, 95-99.
  12. Kang, K. H., B. S. Noh, J. H. Seo, and W. D. Hur. 1998. Food analysis. pp. 109-110, SungKyunkwan University Press. Seoul. Korea.
  13. Kim, J. H., S. Y. Lee, K. B. W. R. Kim, E. J. Song, A. R. Kim, M. J. Kim, K.W. Ji, I. S. Ahn, and D. H. Ahn. 2007. Effects of Glycyrrhiza uralensis, Menthae herba, Schizandra chinensis and chitosan on the self-life and quality of Takju. J. Korean Soc. Food Sci. Nutr. 36, 1436-1443. https://doi.org/10.3746/jkfn.2007.36.11.1436
  14. 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.
  15. Kim, S. and W. Cho. 2006. Effects of Takju (Korean turbid rice wine) Lees on the serum glucose levels in streptozotocin-induced diabetic rats. Korean J. Food Culture 21, 638-643.
  16. Kim, S. K., E. J. Lee, K. Y. Park, and H. K. Jun. 1998. Bacteriocin produced by Lactobacillus curvatus SE1 isolated from Kimchi. J. Microbiol. Biotechnol. 8, 588-594.
  17. Kwan, S. J. and J. H. Sohn. 2011. Analysis of microbial diversity in Nuruk using PCR-DGGE. J. Life Sci. 22, 110-116. https://doi.org/10.5352/JLS.2012.22.1.110
  18. 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.
  19. Lee, J. S., T. S. Lee, S. O. Park, and B. S. Noh. 1996. Flavor components in mash of takju prepared by different raw materials. Korean J. Food Sci. Technol. 28, 316-323.
  20. Lee, J. S., T. S. Lee, B. S. Noh, and S. O. Park. 1996. Quality characteristics of mash of takju prepared by different raw materials. Korean J. Food Sci. Technol. 28, 330-336.
  21. Lee, H. S., K. H. Hong, K. H. Yoon, J. M. Kim, and S. M. Kim. 2009. Effect of Korean turbid rice wine (takju) less extract on blood glucose level in db/db mouse. Korean J. Food Sci. Technol. 23, 109-113.
  22. Lopez, I., F. Ruiz-Larrea, L. Cocolin, E. Orr, T. Phister, M. Marshall, J. Vander Gheynst, and D. A. Mills. 2003. Design and evaluation of PCR primers for analysis of bacterial populations in wine by denaturing gradient gel electrophoresis. Appl. Environ. Microbiol. 69, 6801-6807. https://doi.org/10.1128/AEM.69.11.6801-6807.2003
  23. 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. https://doi.org/10.1002/yea.1124
  24. 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. https://doi.org/10.1128/AEM.69.12.7453-7461.2003
  25. Park, S. S., J. J. Kim, J. A. Yoon, J. H. Lee, B. O. Jung, and S. J. Chung. 2011. Preparation and quality characteristics of Takju (Rice Wine) with Opuntiaficus-indica var., saboten and Chito-oligosaccharide. J. Chitin Chitosan 16, 164-169.
  26. 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. https://doi.org/10.1128/AEM.71.4.1977-1986.2005
  27. 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. https://doi.org/10.1046/j.1365-2583.2003.00390.x
  28. 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. https://doi.org/10.1128/AEM.72.3.2148-2154.2006
  29. Shin, M. O., M. H. Kim, and S. J. Bae. 2010. The Effect of Makgeolli on blood flow, serum lipid improvement and inhibition of ACE in vitro. J. Life Sci. 20, 710-716. https://doi.org/10.5352/JLS.2010.20.5.710
  30. Shin, M. O., D. Y. Kang, M. H. Kim, and S. J. Bae. 2008. Effect of growth inhibition and quinine reductase activity stimulation of Makgeoly fractions in various cancer cells. J. Korean Soc. Food Sci. Nutr. 37, 288-293. https://doi.org/10.3746/jkfn.2008.37.3.288
  31. Seo, M. Y., J. K. Lee, B. H. Ahn, and S. K. Cha. 2005. The changes of microflora during the fermentation of Takju and Yakju. Korean J. Food Sci. Technol. 37, 61-66.
  32. Thompson, J. D., T. J. Gibson, F. Plewniak, F. Jeanmougin, and D. G. Higgins. 1997. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. 25, 4876-4882. https://doi.org/10.1093/nar/25.24.4876
  33. Woo, K. S., J. S. Lee, J. Y. Ko, S. B. Song, B. G. Oh, J. R. Kang, M. H. Nam, I. S. Ryu, and M. C. Seo. 2010. Physicochemical characteristics of Korean traditional wine fermented from foxtail Millet (Setaria italica Beauvios) and nuruk at different addition rates. Korean J. Food Sci. Technol. 42, 298-303.
  34. Woo, K. S., S. B. Song, J. S. Lee, J. Y. Ko, J. R. Kang, B. G. Oh, M. H. Nam, I. S. Ryu, and M. C. Seo. 2010. Physicochemical characteristics of Korean traditional wine made from proso millet (Panicum miliaceum L.) at different addition rates with two kinds of nuruk. Korean J. Crop Sci. 55, 119-125.
  35. 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.
  36. 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. https://doi.org/10.1251/bpo6
  37. Ye, E. J., S. J. Kim, C. H. Park, and M. J. Bae. 2005. Antioxidant and anticancer activities of Ginseng treated with traditional rice wine stream process method. J. Korean Soc. Food Sci. Nutr. 34, 599-604. https://doi.org/10.3746/jkfn.2005.34.5.599
  38. Yu, C. H., S. Y. Hong, and J. S. Koh. 2002. Zymological properties of foztail millet win-making by isolated strain from nuruk. J. Korean Soc. Agric. Chem. Biotechnol. 45, 138-144.

피인용 문헌

  1. Isolation and Characteristics of Fucoidan Degrading Bacterium from Marine vol.22, pp.12, 2012, https://doi.org/10.5352/JLS.2012.22.12.1724
  2. Physicochemical Characteristics and Biological Activities of Makgeolli Supplemented with the Fruit of Akebia quinata during Fermentation vol.45, pp.5, 2013, https://doi.org/10.9721/KJFST.2013.45.5.619
  3. Fermentation Characteristics of Shindari Added with Carrot vol.31, pp.1, 2015, https://doi.org/10.9724/kfcs.2015.31.1.009
  4. Diversity of Lactic Acid Bacteria (LAB) in Makgeolli and Their Production of γ-Aminobutyric Acid vol.47, pp.2, 2015, https://doi.org/10.9721/KJFST.2015.47.2.204
  5. Quality Characteristics and Biogenic Amine Production of Makgeolli Brewed with Commercial Nuruks vol.45, pp.6, 2013, https://doi.org/10.9721/KJFST.2013.45.6.727
  6. Characteristics of Takju (a Cloudy Korean Rice Wine) Prepared with Nuruk (a Traditional Korean Rice Wine Fermentation Starter), and Identification of Lactic Acid Bacteria in Nuruk vol.46, pp.2, 2014, https://doi.org/10.9721/KJFST.2014.46.2.153
  7. Variations of Properties and Microbial Community during Fermentation of Makgeollies by Isolated Yeasts from Traditional Makgeollies vol.23, pp.6, 2013, https://doi.org/10.5352/JLS.2013.23.6.796
  8. Formation of Biogenic Amines by Lactobacillus plantarum Isolated from Makgeolli vol.46, pp.4, 2014, https://doi.org/10.9721/KJFST.2014.46.4.438
  9. Diversity, Saccharification Capacity, and Toxigenicity Analyses of Fungal Isolates in Nuruk vol.42, pp.3, 2014, https://doi.org/10.4489/KJM.2014.42.3.191
  10. Antimicrobial and ACE Inhibitory Activities of Citrus unshiu Fermented with Lactic Acid Bacteria vol.44, pp.7, 2015, https://doi.org/10.3746/jkfn.2015.44.7.1084