Food Science and Preservation (한국식품저장유통학회지)

The Korean Society of Food Preservation (한국식품저장유통학회)
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Expression and Isolation of Limonoid UDP-glucosyltransferase, a Bitterness-reducing Enzyme, in E.coli

감귤의 고미제거 효소인 limonoid UDP-glucosyltransferase의 대장균 내에서의 발현과 이의 분리

  • K.Cho, So-Mi (Faculty of Biotechnology, College of Applied Life Science, Jeju National University) ;
  • Kim, Young-Mee (Department of Medicin, School of Applied Marine Science, Jeju National University) ;
  • Kim, Min-Young (Faculty of Biotechnology, College of Applied Life Science, Jeju National University) ;
  • Lee, Do-Seung (Faculty of Biotechnology, College of Applied Life Science, Jeju National University) ;
  • Kim, Jae-Hoon (Faculty of Biotechnology, College of Applied Life Science, Jeju National University) ;
  • Park, Se-Pill (Faculty of Biotechnology, College of Applied Life Science, Jeju National University) ;
  • Riu, Key-Zung (Faculty of Biotechnology, College of Applied Life Science, Jeju National University) ;
  • Lee, Dong-Sun (Faculty of Biotechnology, College of Applied Life Science, Jeju National University)
  • 김소미 (제주대학교 생명자원과학대학 생명공학부) ;
  • 김영미 (제주대학교 아열대열대생물 유전자 은행센터) ;
  • 김민영 (제주대학교 생명자원과학대학 생명공학부) ;
  • 이도승 (제주대학교 생명자원과학대학 생명공학부) ;
  • 김재훈 (제주대학교 생명자원과학대학 생명공학부) ;
  • 박세필 (제주대학교 생명자원과학대학 생명공학부) ;
  • 류기중 (제주대학교 생명자원과학대학 생명공학부) ;
  • 이동선 (제주대학교 생명자원과학대학 생명공학부)
  • Received : 2010.10.11
  • Accepted : 2011.02.11
  • Published : 2011.04.30
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Abstract

Limonoids are abundant as bitter taste in citrus fruit and other plants. Interestingly. limonoid UDP-glucosyltransferase (LUGT) effectively ameliorates the bitterness from limonoid. The high level of LUGT expression in Escherichia coli can result in the formation of insoluble aggregates known as inclusion bodies. We isolated the soluble LUGT protein when this inclusion body was renaturated with ${\beta}$-cyclidextrin treatment after protein denaturation by urea. Our present results suggest that the isolation of LUGT from inclusion body in cells leads to shed light to characterize the enzyme for food industry purposes.

Limonoid는 항바이러스 및 항균제로써의 치료적인 목적으로 널리 연구되고 있는 성분으로 감귤에서 풍부하게 존재한다. 그러나 성분 자체의 쓴맛으로 인하여 기호성이 저하되므로 이를 해결하는 노력이 필요하며 제품 개발이 요구되고 있는 실정이다. 이러한 쓴맛을 제거할 수 있는 훌륭한 효소로써 LUGT가 주목되고 있으며, 이에 효소의 특성화 연구를 위하여 분리 및 정제를 시도하였다. Plasmid vector인 pET30a(+)을 사용하여 대장균에서 효소 생성 을 위한 최적 발현조건을 검토한 결과, 0.5 mM IPTG 조건에서 $37^{\circ}C$에서 5시간동안 배양하는 것이 최적 과발현 조건이었다. 그러나 세포내 효소 발현은 불용성의 inclusion body로 존재하므로 단백질 변성제인 urea와 재생제인 ${\beta}$-cyclodextrin을 사용하여 순수분리가 가능하였다. 이러한 방법은 LUGT의 당단백질의 특성연구는 물론 이를 이용한 제주 감귤의 고부가가치를 위한 기초연구에 많은 도움을 줄 것으로 판단된다.

Keywords

References

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