The Korean Journal of Mycology (한국균학회지)

The Korean Society of Mycology (한국균학회)
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Optimal Culture Conditions for Penicillium rubefaciens NNIBRFG5039 Possessing Antimicrobial Activity

항균활성 보유 Penicillium rubefaciens NNIBRFG5039의 최적배양 조건

  • Hwang, Hye Jin (Fungi Research Team, Microbial Research Department, Nakdonggang National Institute of Biological Resources (NNIBR)) ;
  • Mun, Hye Yeon (Fungi Research Team, Microbial Research Department, Nakdonggang National Institute of Biological Resources (NNIBR)) ;
  • Hwang, Buyng Su (Animal&Plant Utilization Team, Animal&Plant Research Department, Nakdonggang National Institute of Biological Resources (NNIBR)) ;
  • Nam, Young Ho (Environmental Microbiology Research Team, Microbial Research Department, Nakdonggang National Institute of Biological Resources (NNIBR)) ;
  • Chung, Eu Jin (Environmental Microbiology Research Team, Microbial Research Department, Nakdonggang National Institute of Biological Resources (NNIBR))
  • 황혜진 (국립낙동강생물자원관 균류연구팀) ;
  • 문혜연 (국립낙동강생물자원관 균류연구팀) ;
  • 황병수 (국립낙동강생물자원관 동식물활용연구팀) ;
  • 남영호 (국립낙동강생물자원관 환경미생물연구팀) ;
  • 정유진 (국립낙동강생물자원관 환경미생물연구팀)
  • Received : 2020.01.08
  • Accepted : 2020.03.26
  • Published : 2020.03.31
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Abstract

In screening for antimicrobial activity against methicillin-resistant Staphylococcus aureus (MRSA) KCCM 40510 and Bacillus cereus KCTC 3624, NNIBRFG5039 was isolated from the air in Sangju-si, Gyeongsangbuk-do. Based on a high sequence similarity of the internal transcribed spacer (ITS) region, NNIBRFG5039 was determined to be closely related to Penicillium rubefaciens CBS 139145. The optimal media, initial pH, and temperature for mycelial growth and antimicrobial activity of P. rubefaciens NNIBRFG5039 were determined as follows: potato dextrose broth (PDB), pH 6.5, and 30℃, respectively. Under the optimal culture conditions, maximum mycelial growth (12.4 g L-1) and antibacterial activity (7.5 mm zone of inhibition against MRSA KCCM 40510, and 5.0 mm zone of inhibition against B. cereus KCTC 3624) were observed in a 5 L stirred-tank fermenter. We also isolated the antimicrobial compound from an ethyl acetate fraction, and its chemical structure was identified as (S)-6-hydroxymellein (1) by ESI-MS, 1H-NMR, and 13C-NMR. Consequently, the extract from P. rubefaciens NNIBRFG5039 may be used in functional materials for antimicrobial-related applications.

본 연구에서는 methicillin resistant Staphylococcus aureus subsp. aureus (MRSA) KCCM 40510 및 Bacillus cereus KCTC 3624 균주에 대한 항균활성을 보유한 균류 Penicillium rubefaciens NNIBRFG5039를 경북 상주시 도남동 공기 중으로부터 분리·동정하였고, 배양조건에 따른 균사체 생육 및 항균활성을 비교하였다. 그 결과, P. rubefaciens NNIBRFG5039는 PDB 배지, 배양온도 30℃, 초기 pH 6.5로 배양하였을 때 항균활성이 가장 높게 나타나는 것을 확인하였다. 최적조건하에서 5L fermenter를 이용하여 배양시간에 따른 균사체 건조중량 및 항균활성을 비교한 결과, 배양 5일째에 생장 및 항균활성이 가장 높았다. P. rubefaciens NNIBRFG5039의 배양여액의 항균활성 스펙트럼을 조사한 결과, methicillin-resistant Staphylococcus aureus subsp. aureus CCARM 3089·3090·3091·3095와 KCCM 40510, Bacillus cereus KCTC 3624, B.subtilis KACC 10111, Filobasidium neoformans KCTC 7902, Enterococcus faecalis KCCM 11814에 대해서도 항균활성을 가지는 것을 확인하였다. 또한, P. rubefaciens NNIBRFG5039의 배양여액으로부터 항균활성 후보물질을 각종 크로마토그라피법으로 순수분리하고, NMR과 ESI-MS 등의 기기분석을 실시하여 구조를 (S)-6-hydroxymellein (1)으로 동정하였다.

Keywords

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