Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Optimization of Media Composition on the Production of Melanin Bleaching Enzyme from Peniophora sp. JS17

Peniophora sp. JS17 유래 멜라닌 탈색 효소 생산을 위한 배지 조성의 최적화

  • Son, Min-Jeong (Biomedical Engineering & Biotechnology Major, Division of Applied Bioengineering, Dong-Eui University) ;
  • Kim, Yeon-Hee (Biomedical Engineering & Biotechnology Major, Division of Applied Bioengineering, Dong-Eui University) ;
  • Nam, Soo-Wan (Biomedical Engineering & Biotechnology Major, Division of Applied Bioengineering, Dong-Eui University) ;
  • Jeon, Sung-Jong (Biomedical Engineering & Biotechnology Major, Division of Applied Bioengineering, Dong-Eui University)
  • 손민정 (동의대학교바이오응용공학부의생명공학전공) ;
  • 김연희 (동의대학교바이오응용공학부의생명공학전공) ;
  • 남수완 (동의대학교바이오응용공학부의생명공학전공) ;
  • 전숭종 (동의대학교바이오응용공학부의생명공학전공)
  • Received : 2018.09.14
  • Accepted : 2018.11.17
  • Published : 2019.06.28
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Abstract

Peniphora sp. JS17, isolated from forest old tree, produced extracellular enzymes that decolorized human hair melanin. The JS17 strain had laccase and manganese peroxidase activity while it did not has lignin peroxidase activity. Batch culture indicated that the melanin decolorization activity of JS17 strain originated from laccase. The culture conditions to maximize the production of melanin bleaching enzymes from Peniophora sp. JS17 mycelia were investigated. Among the tested media for the laccase production, minimal medium (2% glucose, 0.2% malt extract, 0.1% $KH_2PO_4$, 0.4% $MgSO_4{\cdot}7H_2O$) showed the highest activity of laccase. Then, to optimize the culture condition for the laccase activity, the influence of various carbon and nitrogen sources was investigated in minimal medium. Among various carbon and nitrogen sources, 2% xylose and 0.4% tryptone showed the highest production of laccase, respectively. The enzyme was purified using $(NH_4)_2SO_4$ precipitation and Hitrap Q sepharose column, and the purified enzyme showed two isoenzymatic bands with molecular masses of about 70 kDa by SDS-PAGE. The melanin decolorization activity was 77% and 55% within 48 h in the presence of 1-hydroxybenzotriazole (HBT) and syringaldehyde, respectively, whereas only about 9% melanin decolorized in case of no mediator.

삼림지역의 고목에서 분리한 Peniophora sp. JS17 균주는 사람 머리카락 유래 멜라닌을 탈색하는 세포 외 분비효소를 생산했다. JS17 균주는 laccase 및 manganese peroxidase 활성을 나타냈지만 lignin peroxidase 활성은 나타내지 않았다. 본 균주를 회분배양한 결과 멜라닌 탈색 활성은 laccase 활성으로부터 유래하는 것으로 확인되었다. Peniophora sp. JS17 균사체로부터 멜라닌 탈색 효소를 생산하기 위한 배지 조건을 조사하였다. 다양한 합성 배지 중에서 minimal medium (2% glucose, 0.2% malt extract, 0.1% $KH_2PO_4$, 0.4% $MgSO_4{\cdot}7H_2O$)이 가장 높은 laccase 활성을 나타내었다. Laccase 생산에 대한 배양 조건을 최적화하기 위하여 minimal medium의 조성 중에서 탄소원 및 질소원에 대한 영향을 조사하였다. 다양한 탄소원 및 질소원 중에서 각각 2% xylose 및 0.4% tryptone의 경우에 가장 높은 laccase 활성을 나타내었다. Ammonium sulfate 침전 및 Hitrap Q Sepharose column을 이용하여 정제한 효소는 SDS-PAGE에서 약 70 kDa의 분자량 부근에 2종류의 isozyme 형태로 존재 하였다. 멜라닌 탈색율은 HBT 및 syringaldehyde의 존재하에서 48시간 만에 각각 77% 및 55%를 나타내었고, mediator가 없는 조건에서는 9%의 멜라닌 탈색율을 나타내었다.

Keywords

References

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