메주와 누룩은 한국 전통 발효 식품에 사용되는 스타터로, Aspergillus속 곰팡이나 aflatoxin에 노출되기 쉽다. 본 연구에서는 우리나라에서 시판되는 57개의 메주 시료와 18개의 누룩 시료로부터 Aspergillus 속 곰팡이를 분리하고 동정하였다. 분리주의 aflatoxin 생성 가능성을 평가하기 위하여 multiplex PCR을 통해 aflatoxin 생합성 유전자 (aflO, aflP, aflR)를 확인하고, 이들 분리주에 의해 생성되는 aflatoxin 함량을 HPLC로 조사하였다. 뿐만 아니라 시판 메주와 누룩 시료 중 aflatoxin 함량을 분석하였다. 그 결과, 메주 시료로부터 130개, 누룩 시료로부터 47개 균주가 분리되어 총 177개의 분리주를 확인 및 동정하였다. 각 각 메주와 누룩으로부터 분리된 19.2% (25/130), 10.6% (5/ 47)의 분리주가 3 종류의 aflatoxin 생합성 유전자를 모두 보유하였으며, 그 중 메주로부터 분리된 5개의 분리주가 실제 로 aflatoxin을 생성하였다. 시판 메주와 누룩 시료 중 aflatoxin 함량을 분석한 결과, 88% (51/58)의 메주 시료의 aflatoxin 오염 수준은 모두 검출한계 미만으로 나타났고, 누룩 또한 시료의 39% (7/18)가 검출한계 미만으로 확인되었다. 메주와 누룩에서 분리된 분리주 중 aflatoxin 생합성 유전자를 모두 보유하거나 배지 상에서 aflatoxin 생성을 보여준 aflatoxigenic 균주는 존재하였으나 유통되고 있는 시료에서 aflatoxin 오염 빈도는 낮은 수준임을 확인할 수 있었다.

Meju and nuruk (respectively soybean and malt) are traditional Korean fermentation starters that are vulnerable to contamination by harmful microorganisms such as aflatoxigenic fungi and their associated aflatoxins (AFs). In this study, Aspergillus spp. were isolated and identified from a total of 57 meju and 18 nuruk samples collected from Korean markets. Their potential aflatoxigenicity was investigated by examining the presence of three aflatoxin biosynthetic genes (aflO, aflP, and aflR) using multiplex polymerase chain reaction (mPCR) assays. Thereafter, aflatoxin production of isolates and the natural occurrence of AFs in meju and nuruk samples were analyzed by high-performance liquid chromatography (HPLC). A total of 177 Aspergillus isolates were identified and 130 isolates were obtained from meju samples. Of these, 25 isolates (19.2%) contained all three aflatoxin biosynthetic genes, and five (20%) of these isolates produced aflatoxins. Forty-seven of the Aspergillus isolates were obtained from nuruk samples, five of which (10.6%) expressed all three AF biosynthetic genes; however, none of these strains produced AFs. HPLC analysis showed that 88% (51/58) of the meju samples and 39% (7/18) of nuruk samples were not contaminated with AFs (below limit of detection). Among the isolates isolated from meju and nuruk, there were aflatoxigenic strains containing all three aflatoxin biosynthetic genes or producing aflatoxin in medium, but the frequency of aflatoxin contamination was low in the meju and nuruk samples.

ABSTRACT
Materials and Methods
    Sample collection
    Isolation and identification of Aspergillus speciesfrom Meju and Nuruk samples
    Aflatoxigenicity analysis using mPCR
    Aflatoxin extraction from fungal culture
    Aflatoxin extraction from Meju and Nuruk samples
    Aflatoxin detection using HPLC-FLD
    Aflatoxin confirmation by LC-tandem mass spectrometry(LC-MS/MS)
Results
    Isolation and identification of Aspergillus isolatesfrom Meju and Nuruk
    Aflatoxigenicity analysis of Aspergillus isolates usingmPCR
    Occurrence of aflatoxins in Nuruk and Meju samples
Discussion
국문요약
References

• So Young Woo(Advanced Food Safety Research Group, BK21 Plus, School of Food Science and Technology, Chung-Ang University)
• Sang Yoo Lee(Advanced Food Safety Research Group, BK21 Plus, School of Food Science and Technology, Chung-Ang University)
• Fei Tian(Advanced Food Safety Research Group, BK21 Plus, School of Food Science and Technology, Chung-Ang University)
• A-Yeong Jeong(Advanced Food Safety Research Group, BK21 Plus, School of Food Science and Technology, Chung-Ang University)
• Cha Nee Yoo(Advanced Food Safety Research Group, BK21 Plus, School of Food Science and Technology, Chung-Ang University)
• Seung Yoon Kang(Advanced Food Safety Research Group, BK21 Plus, School of Food Science and Technology, Chung-Ang University)
• Hyang Sook Chun(Advanced Food Safety Research Group, BK21 Plus, School of Food Science and Technology, Chung-Ang University) Correspondence to