Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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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 (Fermentation & Food Processing Division, Department of Agro-food Resource, NAAS, RDA) ;
  • Lee, You-Jung (Fermentation & Food Processing Division, Department of Agro-food Resource, NAAS, RDA) ;
  • Yeo, Soo-Hwan (Fermentation & Food Processing Division, Department of Agro-food Resource, NAAS, RDA) ;
  • Choi, Hye-Sun (Fermentation & Food Processing Division, Department of Agro-food Resource, NAAS, RDA) ;
  • Park, Hye-Young (Fermentation & Food Processing Division, Department of Agro-food Resource, NAAS, RDA) ;
  • Park, Heui-Dong (Department of Food Science & Technology, Agro Biotechnology Education Center, Kyungpook National University) ;
  • Baek, Seong-Yeol (Fermentation & Food Processing Division, Department of Agro-food Resource, NAAS, RDA)
  • 윤혜주 (농촌진흥청 국립농업과학원 농식품자원부 발효이용과) ;
  • 이유정 (농촌진흥청 국립농업과학원 농식품자원부 발효이용과) ;
  • 여수환 (농촌진흥청 국립농업과학원 농식품자원부 발효이용과) ;
  • 최혜선 (농촌진흥청 국립농업과학원 농식품자원부 발효이용과) ;
  • 박혜영 (농촌진흥청 국립농업과학원 농식품자원부 발효이용과) ;
  • 박희동 (경북대학교 농업생명과학대학 식품공학과) ;
  • 백성열 (농촌진흥청 국립농업과학원 농식품자원부 발효이용과)
  • Received : 2012.05.10
  • Accepted : 2012.06.13
  • Published : 2012.06.28
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Abstract

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.

경기도 일대에서 수집한 메주 시료에서 지질분해 활성을 나타내는 균주 Y124를 분리하여 동정한 결과 Yarrowia lipolytica와 100% 상동성을 보였다. 분리 균주가 생산하는 lipase의 조효소에 대한 일반적인 특성을 조사한 결과, 탄소원으로 olive oil을 단독으로 사용한 YPO 배지에서 8시간 배양하였을 때 lipase 활성이 가장 높게 나타났다. YPD 배지에서는 lipase 활성이 거의 없었으며, olive oil과 glucose를 모두 포함하는 YPDO 배지에서는 lipase 활성이 YPO 배지 보다 낮았다. 그리고 olive oil 농도에 따른 lipase 활성을 측정한 결과, olive oil 무첨가보다 0.7% 첨가하여 8시간 배양했을 때 lipase 활성이134 U/mL으로 가장 높게 나타나 lipase의 생산이 olive oil의 첨가에 의해 유도되는 것으로 생각된다. 생육온도에 따른 lipase 활성 측정한 결과, $30^{\circ}C$에 배양하였을 때 배양 8시간에 가장 높은 활성이 나타났고, $25^{\circ}C$와 $37^{\circ}C$에 배양하였을 때는 배양 12시간에 활성이 가장 높게 나타났으며, Y124균주의 lipase 활성 최적 온도는 $30^{\circ}C$로 나타났다. 그리고 lipase의 기질 친화도를 확인한 결과 Y124균주가 생산하는 lipase의 경우 p-nitrophenyl octanoate ($C_8$)에서 가장 높은 활성이 나타났다.

Keywords

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