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Fungi associated with the traditional starter cultures used for rice wine in Korea

  • Food Science/Microbiology
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Abstract

The ‘Nuruk’ starter culture has been used for many years in the fermentation of rice wine in Korea. In present study, mycobiota in Nuruk cultures was identified by performing morphological, physiological, and phylogenetical analyses. Mucorales was the most common mycobiota in Nuruk, followed by yeast and Aspergillus. The composition of fungal species in Nuruk was different among samples and did not correlate with the geographical location from where the Nuruk culture was manufactured. For more detailed identification, 174 filamentous fungal strains were isolated from 39 Nuruk samples. Although the morphological and molecular analyses showed that the strains were identical at the genus level, some discordance was identified between species. Of the 174 strains, 160 showed thermotolerance, and the level of thermotolerance matched the clade generated by phylogenetic analysis. Six genera (Lichtheimia, Aspergillus, Rhizopus, Rhizomucor, Mucor, and Syncephalastrum) and 17 fungal species were identified. Among the genera, the genus Syncephalastrum had not been previously identified in Nuruk cultures, and the isolate was identified as Syncephalastrum racemosum. Two genera, Lichtheimia and Aspergillus, comprised approximately 84% of the filamentous fungal isolates from the Nuruk samples, and Lichtheimia ramosa and Aspergillus oryzae were the most commonly found species. The controversy regarding the presence of mycobiota in Nuruk starter cultures was addressed, and results showed that Nuruk contains unique mycobiota not yet found in other Asian starter cultures.

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References

  • Alvarez E, Sutton DA, Cano J, Fothergill AW, Stchigel A, Rinaldi MG, and Guarro J (2009) Spectrum of zygomycete species identified from clinically significant specimens in the United States. J Clin Microbiol 47, 1650–1656.

    CAS  Google Scholar 

  • Assante G, Camarda L, Locci R, Merlini L, Nasini G, and Papadopoulos E (1981) Isolation and structure of red pigments from Aspergillus flavus and related species, grown on a differential medium. J Agric Food Chem 29, 785–787.

    CAS  Google Scholar 

  • Bae KH, Shin KS, Ryu HY, Kwon CS, and Sohn HY (2007) Identification and fermentation characteristics of lactic acid bacteria isolated from the fermentation broth of Korean traditional liquor, Andong-Soju. Korean J Microbiol Biotechnol 35, 310–315. (In Korean).

    CAS  Google Scholar 

  • Balajee SA, Borman AM, Brandt ME, Cano J, Cuenca-Estrella M, Dannaoui E, Guarro J, Haase G, Kibbler CC, Meyer W, O’Donnell K, Petti CA, Rodriguez-Tudela JL, Sutton D, Velegraki A, and Wickes BL (2009) Sequence-based identification of Aspergillus, Fusarium, and Mucorales species in the clinical mycology laboratory: Where are we and where should we go from here? J Clin Microbiol 47, 877–884.

    CAS  Google Scholar 

  • Bothast RJ and Fennell DI (1974) A medium for rapid identification and enumeration of Aspergillus flavus and related organisms. Mycologia 66, 365–369.

    CAS  Google Scholar 

  • Ciesla MC, Kammeyer PL, Yeldandi V, Petruzzelli GJ, and Yong SL (2000) Identification of the asexual state of Rhizopus species on histologic tissue sections in a patient with rhinocerebral mucormycosis. Arch Pathol Lab Med 124, 883–887.

    CAS  Google Scholar 

  • de Hoog GS and Guarro J (2000) In Atlas of Clinical Fungi, Centraalbureauvoor Schimmelcultures, Baarn, The Netherlands.

    Google Scholar 

  • Garcia-Hermoso D, Hoinard D, Gantier J-C, Grenouillet F, Dromer F, and Dannaoui E (2009) Molecular and phenotypic evaluation of Lichtheimia corymbifera (formerly Absidia corymbifera) complex isolates associated with human mucormycosis: Rehabilitation of L. ramosa. J Clin Microbiol 47, 3862–3870.

    CAS  Google Scholar 

  • Geiser DM, Klich MA, Frisvad JC, Peterson SW, Varga J, and Samson RA (2007) The current status of species recognition and identification in Aspergillus. Stud Mycol 59, 1–10.

    CAS  Google Scholar 

  • Glass NL and Donaldson GC (1995) Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes. Appl Environ Microbiol 61, 1323–1330.

    CAS  Google Scholar 

  • Guarro J, Gene J, and Stchigel AM (1999) Developments in fungal taxonomy. Clin Microbiol Rev 12, 454–500.

    CAS  Google Scholar 

  • Ha DM, Kim DC, Hong SM, and Lee CW (1989) Identification and properties of starch utilizing yeasts isolated from Nuruk. J Korean Agric Chem Soc 32, 408–415. (In Korean).

    CAS  Google Scholar 

  • Han EH, Lee TS, Noh BS, and Lee DS (1997) Quality characteristics in mash of takju prepared by using different Nuruk during fermentation. Korean J Food Sci Technol 29, 555–562. (In Korean).

    Google Scholar 

  • Henry T, Iwen PC, and Hinrichs SH (2000) Identification of Aspergillus species using internal transcribed spacer regions 1 and 2. J Clin Microbiol 38, 1510–1515.

    CAS  Google Scholar 

  • Hesseltine CW (1983) Microbiology of oriental fermented foods. Ann Rev Microbiol 37, 575–601.

    CAS  Google Scholar 

  • Hoffmann K, Discher S, and Voigt K (2007) Revision of the genus Absidia (Mucorales, Zygomycetes) based on physiological, phylogenetic, and morphological characters; thermotolerant Absidia spp. form a coherent group, Mycocladiaceae fam. nov. Mycol Res 111, 1169–1183.

    CAS  Google Scholar 

  • Hoffmann K, Walther G, and Voigt K (2009) Mycocladus vs. Lichtheimia: A correction (Lichtheimiaceae fam. nov., Mucorales, Mucoromycotina). Mycol Res 113, 277–278.

    Google Scholar 

  • Jeyaram K, Singh WM, Capece A, and Romano P (2008) Molecular identification of yeast species associated with ‘Hamei’ — A traditional starter used for rice wine production in Manipur, India. Int J Food Microbiol 124, 115–125.

    CAS  Google Scholar 

  • Jo GY and Lee CW (1997) Isolation and identification of the fungi from Nuruk. J Korean Soc Food Sci Nutr 26, 759–766. (In Korean).

    Google Scholar 

  • Jo KY and Ha DM (1995) Isolation and identification of the lactic acid bacteria from Nuruk. Agr Chem Biotechnol 38, 95–99. (In Korean).

    Google Scholar 

  • Kim HR, Baek SH, Seo MJ, and Ahn BH (2006) Feasibility of Cheonghju brewing with wild type yeast strains from Nuruks. Korean J Microbiol Biotechnol 34, 244–249. (In Korean).

    CAS  Google Scholar 

  • Kim HS, Hyun JS, Kim J, Ha HP, and Yu TS (1997) Characteristics of useful fungi isolated from traditional Korean Nuruk. J Korean Soc Food Sci Nutr 26, 767–774. (In Korean).

    Google Scholar 

  • Kim HS, Hyun JS, Kim J, Ha HP, and Yu TS (1998) Enzymological characteristics and identification of useful fungi isolated from traditional Korean Nuruk. Korean J Microbiol Biotechnol 26, 456–464. (In Korean).

    CAS  Google Scholar 

  • Kim JY and Koh JS (2004) Screening of brewing yeasts and saccharifying molds for foxtail millet-wine making. Korean Soc Appl Biol Chem 47, 78–84. (In Korean).

    CAS  Google Scholar 

  • Klich MA (2002) In Identification of Common Aspergillus Species, Centraalbureauvoor Schimmelcultures, Utrecht, The Netherlands.

    Google Scholar 

  • Kozaki M and Uchimura T (1990) Microorganisms in Chinese starter “Bubod” and rice wine “Tapuy” in the Philippines. J Brew Soc Jpn 85, 818–824. (In Japanese).

    Google Scholar 

  • Lee HH, Lee JH, Ko YJ, Park MH, Lee JO, and Ryu CH (2009) Changes in allergenicity and quality of Nuruk during fermentation. J Korean Soc Food Sci Nutr 38, 76–82. (In Korean).

    CAS  Google Scholar 

  • Lee JH and Yu TS (2000) Identification and characteristics of lactic acid bacteria isolated from Nuruk. Korean J Biotechnol Bioeng 15, 359–365. (In Korean).

    Google Scholar 

  • Lee SJ and Ahn BH (2010) Sensory profiling of rice wines made with Nuruks using different ingredients. Korean J Food Sci Technol 42, 119–123. (In Korean).

    Google Scholar 

  • Lee SS, Kim KS, Eom AH, Sung CK, and Hong IP (2002) Production of Korean traditional rice-wines made from cultures of the single fungal isolates under laboratory conditions. Korean J Mycol 30, 61–65. (In Korean)

    Google Scholar 

  • Lee WK, Kim JR, and Lee MW (1987) Studies on the changes in free amino acids and organic acids of Takju prepared with different Koji strains. J Korean Agric Chem Soc 30, 323–327. (In Korean).

    Google Scholar 

  • Leslie JF and Summerell BA (2006) In The Fusarium Laboratory Manual, Blackwell Pub., Ames, IA.

    Book  Google Scholar 

  • Nikkuni S, Karki TB, Terao T, and Suzuki C (1996) Microflora of Mana, a Nepalese rice koji. J Ferment Bioeng 81, 168–170.

    CAS  Google Scholar 

  • Park JW, Lee KH, and Lee CY (1995) Identification of filamentous molds isolated from Korean traditional Nuruk and their amylolytic activities. Korean J Appl Microbiol Biotechnol 23, 737–746. (In Korean).

    Google Scholar 

  • Pitt JI, Hocking AD, and Glenn DR (1983) An improved medium for the detection of Aspergillus flavus and A. parasiticus. J Appl Microbiol 54, 109–114.

    CAS  Google Scholar 

  • Ribes JA, Vanover-Sams CL, and Baker DJ (2000) Zygomycetes in human disease. Clin Microbiol Rev 13, 236–301.

    CAS  Google Scholar 

  • Rodrigues P, Venâncio A, Kozakiewicz Z, and Lima N (2009) A polyphasic approach to the identification of aflatoxigenic and non-aflatoxigenic strains of Aspergillus Section Flavi isolated from Portuguese almonds. Int J Food Microbiol 129, 187–193.

    CAS  Google Scholar 

  • Saitou N and Nei M (1987) The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol Biol Evol 4, 406–425.

    CAS  Google Scholar 

  • Schipper MAA (1984) A revision of the genus Rhizopus. I. The Rhizopus stolonifer-group and Rhizopus oryzae. Stud Mycol 25, 1–34.

    Google Scholar 

  • Schwarz P, Bretagne S, Gantier J-C, Garcia-Hermoso D, Lortholary O, Dromer F, and Dannaoui E (2006) Molecular identification of zygomycetes from culture and experimentally infected tissues. J Clin Microbiol 44, 340–349.

    CAS  Google Scholar 

  • Shon SK, Rho YH, Kim HJ, and Bae SM (1990) Takju brewing of uncooked rice starch using Rhizopus koji. Korean J Appl Microbiol Biotechnol 18, 506–510. (In Korean).

    Google Scholar 

  • So MH (1995) Aptitudes for Takju brewing of wheat flour- Nuluks made with different mold species. Korean J Food Nutr 8, 6–12. (In Korean).

    Google Scholar 

  • So MH (1999) Characteristics of a modified Nuruk made by inoculation of traditional Nuruk microorganisms. Korean J Food Sci Technol 12, 219–225. (In Korean).

    Google Scholar 

  • Tamura K, Dudley J, Nei M, and Kumar S (2007) MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24, 1596–1599.

    CAS  Google Scholar 

  • Tamura K, Nei M, and Kumar S (2004) Prospects for inferring very large phylogenies by using the neighbor-joining method. Proc Natl Acad Sci USA 101, 11030–11035.

    CAS  Google Scholar 

  • Thanh VN, Mai LT, and Tuan DA (2008) Microbial diversity of traditional Vietnamese alcohol fermentation starters (banh men) as determined by PCR-mediated DGGE. Int J Food Microbiol 128, 268–273.

    CAS  Google Scholar 

  • Thompson JD, Higgins DG, and Gibson TJ (1994) CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positionspecific gap penalties and weight matrix choice. Nucleic Acids Res 22, 4673–4680.

    CAS  Google Scholar 

  • Uchimura T, Takagi S, Watanabe K, and Kozaki M (1990) Absidia sp. in the Chinese starter (Nuruk) in Korea. J Brew Soc Jpn 85, 888–894. (In Japanese).

    Google Scholar 

  • White TJ, Bruns T, Lee S, and Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In PCR Protocols a Guide to Methods and Applications, Innis MA, Gelfand DH, Sninsky JJ, and White TJ (eds.), Academic Press, New York, NY.

    Google Scholar 

  • Wood BJB (1977) Oriental food uses of Aspergillus. In The British Mycological Symposium Series No 1. Genetics and Physiology of Aspergillus, Academic Press, London, UK.

    Google Scholar 

  • Xie G, Li W, Lu J, Cao Y, Fang H, Zou H, and Hu Z (2007) Isolation and identification of representative fungi from Shaoxing rice wine wheat Qu using a polyphasic approach of culture-based and molecular-based methods. J Inst Brew 113, 272–279.

    CAS  Google Scholar 

  • Yang JY and Lee KH (1996) Shelf-life and microbiological study of Sansung Takju. Korean J Food Sci Technol 28, 779–785 (In Korean).

    Google Scholar 

  • Yokoyama N, Tanaka K, Du L, Aramaki I, Kizaki Y, Kobayashi S, and Okazaki N (1994) Microflora and enzyme activities of Chinese Qu. J Brew Soc Jpn 89, 72–76. (In Japanese).

    CAS  Google Scholar 

  • Yu CH, Hong SY, and Koh JS (2002) Zymological properties of foxtail millet wine making by isolated strains from Nuruk. J Korean Agric Chem Soc 45, 138–144. (In Korean).

    CAS  Google Scholar 

  • Yu TS, Kim HS, Hong J, Ha HP, Kim TY, and Yoon IW (1996) Bibliographical study on microorganisms of Nuruk (Until 1945). J Korean Soc Food Sci Nutr 25, 170–179. (In Korean).

    Google Scholar 

  • Yu TS, Kim J, Kim HS, Hyun JS, Ha HP, and Park MG (1998) Bibliographical study on microorganisms of traditional Korean Nuruk (since 1945). J Korean Soc Food Sci Nutr 27, 789–799. (In Korean).

    Google Scholar 

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Correspondence to Yin-Won Lee.

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Yang, S., Lee, J., Kwak, J. et al. Fungi associated with the traditional starter cultures used for rice wine in Korea. J Korean Soc Appl Biol Chem 54, 933–943 (2011). https://doi.org/10.1007/BF03253183

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