• Title/Summary/Keyword: ITS rDNA, Meju

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Use of Restriction Fragment Length Polymorphism Analysis to Differentiate Fungal Strains in Sunchang Meju

  • Jung, Jong-Hyun;Seo, Dong-Ho;Bhoo, Sung-Hee;Ha, Suk-Jin;Kim, Jong-Sang;Kim, Jeong-Hwan;Kwon, Dae-Young;Cha, Jae-Ho;Park, Cheon-Seok
    • Food Science and Biotechnology
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    • v.17 no.4
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    • pp.888-891
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    • 2008
  • Twenty-three fungal strains were isolated from meju that had originated from the Sunchang province, the famous location for making fermented soybean foods in Korea. The restriction fragment length polymorphism (RFLP) of the internal transcribed spacer (ITS) region of the rDNA (ITS-RFLP) was applied to differentiate the isolated fungal strains. First, the ITS region by polymerase chain reaction (PCR) with specific primers was amplified and then cleaved the products with different restriction enzymes. Cleavage of the amplified fragments with the restriction enzymes AluI, HaeIII, HhaI, and TaqI revealed extensive polymorphisms. The ITS-RFLP results highly correlated with ITS sequence analysis. All of the 23 fungal strains were classified into 5 groups by ITS-RFLP analysis. Aspergillus oryzae was the major fungal strain isolated from Sunchang meju (12 out of 23), while Aspergillus fumigatus was the next most frequently isolated strain (7 out of 23). In contrast, it was found that Fusarium asiaticum, Aspergillus sydowii, and Arthrinium sp. were the minor fungal strains in meju.

Aspergillus cumulatus sp. nov., from Rice Straw and Air for Meju Fermentation

  • Kim, Dae-Ho;Kim, Seon-Hwa;Kwon, Soon-Wo;Lee, Jong-Kyu;Hong, Seung-Beom
    • Journal of Microbiology and Biotechnology
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    • v.24 no.3
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    • pp.334-336
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    • 2014
  • A new species named Aspergillus cumulatus sp. nov. is described in Aspergillus section Aspergillus (Eurotium state). The type strain (KACC $47316^T$) of this species was isolated from rice straw used in meju fermentations in Korea, and other strains were isolated from the air in a meju fermentation room. The species is characterized by growth at a wide range of water activities and the formation of aerial hyphae on malt extract 60% sucrose agar (ME60S) that resemble a cumulus cloud. Furthermore, A. cumulatus produces yellow ascomata containing small lenticular ascospores (5.1-5.7 ${\mu}m$) with a wide furrow, low equatorial crests, and tuberculate convex surface. The species is phylogenetically distinct from the other reported Aspergillus section Aspergillus species based on multilocus sequence typing using rDNA-ITS, ${\beta}$-tubulin, calmodulin, and RNA polymerase II genes.

Identification and Characterization of Eurotium rubrum Isolated from Mejuin Korea

  • Yun, Yeo-Hong;Hyun, Min-Woo;Suh, Dong-Yeon;Kim, Yong-Min;Kim, Seong-Hwan
    • Mycobiology
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    • v.37 no.4
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    • pp.251-257
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    • 2009
  • We isolated and identified a strain of Eurotium rubrum from Meju that has not been reported in Korea. This fungus is yellowish brown; reverse dark brown on CYA and PDA while yellow on 2% MEA at $25{^{\circ}C}$. Cleistothecia are first bright yellow and gradually turned brown. Mycerial growth on CYA attained a diameter of 30 mm at $20{^{\circ}C}$, 37 mm at $25{^{\circ}C}$ and 32 mm at $30{^{\circ}C}$ after 15 days. The isolate grew slower on 2% MEA ($<$ 20 mm 15 days at $25{^{\circ}C}$) compared to CYA and PDA ($<$ 40 mm 15 days at $25{^{\circ}C}$). Cleistothecia are superficial, yellow to light brown, globose to subglobose, 40~75 ${\mu}m$ in diameter. Asci are 8-spored and globose to subglobose 8~11 ${\mu}m$. Ascospores are disciform, 4.0~5.0 ${\mu}m$ in length and 4.2~4.5 ${\mu}m$ in width. Conidia are ovate or bacillar, finely roughened to densely spinulose, 4.6~6.0 ${\mu}m$ in length and 3.0~4.3 ${\mu}m$ in width. Compared to known Eurotium rubrum, the Korean isolate showed 99% sequence similarity in ITS rDNA (554 bp) and calmodulin (750 bp) gene and 100% in $\beta$-tubulin (1016 bp) gene. The E. rubrum isolate also had weak $\beta$-glucosidase and protease activities.

Taxonomic Observations of the Isolates of Rhizopus species Inhabiting at the Raw Materials for Korean Traditional Fermented Foods in Korea (한국 전통 발효식품의 원료에서 채집된 Rhizopus속의 분류학적 관찰)

  • Lee, Sang-Sun;Yoon, Young-Sil;Yu, Kee-Won;Sung, Chang-Kun
    • The Korean Journal of Mycology
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    • v.26 no.4 s.87
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    • pp.478-486
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    • 1998
  • The isolates of Rhizopus collected from Korean traditional fermented food meju and nuruk, were compared with the well known species of Rhizopus donated. The isolates of Rhizopus were identified with the numerical analyses calculated from RAPD-DNA bands, and confirmed with the microscopic observations of morphological features on PDA. The isolates of R. oryzae purchased were segregated through the results of RAPD or the morphological features. The species of R. nigricans, known as an illegitimate species, were different from those of R. stolonife and it is estimated that they are species of Rhizopus, again. According to microscopic observations and the result of RAPD, Some isolates of R. oryzae purchased belong to R. nigricans and are completely different from R. oryzae in their growth. The isolates of R. nigricans included with several isolates of R. oryzae identified for the different phenotypes and were heterogenous. The isolate of R. oligosporus were speculated to be different from morphological features of Rhizopus, but to be similar to the species of Absidia on the apophysis of sporangium. Its sporangiophore or mycelium was observed to be dark black, but the sporangia were not in those of R. oligosporus. The isolates collected from Korean traditional nuruk showed genetic diversity, and also considered to be different tastes in Korean rice wines.

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Isolation and Identification of Fungi from a Meju Contaminated with Aflatoxins

  • Jung, Yu Jung;Chung, Soo Hyun;Lee, Hyo Ku;Chun, Hyang Sook;Hong, Seung Beom
    • Journal of Microbiology and Biotechnology
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    • v.22 no.12
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    • pp.1740-1748
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    • 2012
  • A home-made meju sample contaminated naturally with aflatoxins was used for isolation of fungal strains. Overall, 230 fungal isolates were obtained on dichloran rosebengal chloramphenicol (DRBC) and dichloran 18% glycerol (DG18) agar plates. Morphological characteristics and molecular analysis of a partial ${\beta}$-tubulin gene and the internal transcribed spacer (ITS) of rDNA were used for the identification of the isolates. The fungal isolates were divided into 7 genera: Aspergillus, Eurotium, Penicillium, Eupenicillium, Mucor, Lichtheimia, and Curvularia. Three strains from 56 isolates of the A. oryzae/flavus group were found to be aflatoxigenic A. flavus, by the presence of the aflatoxin biosynthesis genes and confirmatory aflatoxin production by high-performance liquid chromatography (HPLC). The predominant isolate from DRBC plates was A. oryzae (42 strains, 36.2%), whereas that from DG18 was A. candidus (61 strains, 53.5%). Out of the 230 isolates, the most common species was A. candidus (34.3%) followed by A. oryzae (22.2%), Mucor circinelloides (13.0%), P. polonicum (10.0%), A. tubingensis (4.8%), and L. ramosa (3.5%). A. flavus and E. chevalieri presented occurrence levels of 2.2%, respectively. The remaining isolates of A. unguis, P. oxalicum, Eupenicillium cinnamopurpureum, A. acidus, E. rubrum, P. chrysogenum, M. racemosus, and C. inaequalis had lower occurrence levels of < 2.0%.

Probiotic Potential of Indigenous Bacillus sp. BCNU 9028 Isolated from Meju (메주로부터 분리한 토착 Bacillus sp. BCNU 9028의 프로바이오틱스로서 이용 가능성)

  • Shin, Hwa-Jin;Bang, Ji-Hun;Choi, Hye-Jung;Kim, Dong-Wan;Ahn, Cheol-Soo;Jeong, Young-Kee;Joo, Woo-Hong
    • Journal of Life Science
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    • v.22 no.5
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    • pp.605-612
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    • 2012
  • Spore-forming bacteria are being used as probiotic supplements for human and animal use, due to their low pH stability and ability to survive the gastric barrier. In this study, the BCNU 9028 strain was screened from meju, a Korean fermented soybean food starter. Biochemical and physiological characteristics, as well as 16S rDNA sequence analyses, indicate that this strain belongs to the genus $Bacillus$. $Bacillus$ sp. BCNU 9028 showed a 92% survivability at pH 2.5 and could also withstand 0.3% ox bile. Furthermore, it was postulated that $Bacillus$ sp. BCNU 9028 could prevent biofilm formation and adherence of food-borne pathogens such as $Listeria$ $monocytogenes$, $S.$ $aureus$ and $E.$ $coli$ on the basis of its autoaggregation and coaggregation capacity with food-borne pathogens. It was shown that BCNU 9028 has good abilities to adhere to the intestinal tract from its hydrophobic character (63.3%). The $Bacillus$ sp. BCNU 9028 strain especially elicited antibacterial activity against both Gram-positive and -negative pathogens. These findings suggested that the $Bacillus$ sp. BCNU 9028 strain could be used as a potential probiotic.

The Isolation and Culture Characterization of a Lipolytic Enzyme Producing Strain from Meju (메주로부터 지질분해 효소 생산 균주의 분리 및 배양학적 특성)

  • Yun, Hye-Ju;Lee, You-Jung;Yeo, Soo-Hwan;Choi, Hye-Sun;Park, Hye-Young;Park, Heui-Dong;Baek, Seong-Yeol
    • Microbiology and Biotechnology Letters
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    • v.40 no.2
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    • pp.98-103
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    • 2012
  • For screening of useful enzymes producing microorganisms from Meju, we isolated high lipase producing strains and their lipolytic enzyme activities were then tested. The lipolytic enzyme activities of isolated microorganisms were therefore tested on the Y124 strain. The gene sequence analysis of ITS from Y124 strain revealed Yarrowia lipolytica. Lipase production by the Y124 strain was studied in media containing various carbon sources. The Y124 strain drastically increased lipolytic enzyme activity in YPO media containing olive oil, as well as in YPDO media containing both olive oil and glucose. Maximal lipase production was achieved in YPD (yeast extract-peptone-D-glucose) media containing 0.7% olive oil when cultured at $30^{\circ}C$ for 8 hrs. The lipase produced from the Y124 strain showed the highest activity in p-NPO (p-nitrophenyl octanoate ($C_8$)), amongst the various p-nitrophenyl esters.