한국미생물·생명공학회지 (Microbiology and Biotechnology Letters)

  • 제36권4호
  • /
  • Pages.299-306
  • /
  • 2008
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  • 1598-642X(pISSN)
  • /
  • 2234-7305(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
권호 표지

비만 억제를 위한 췌장 리파아제 도메인에 대한 특이 난황항체의 개발

Development of Egg Yolk Antibody Specific to the Pancreatic Lipase Domain for Anti-Obesity

  • 우승은 ((주)단바이오텍 생명과학연구소) ;
  • 권진혁 ((주)단바이오텍 생명과학연구소) ;
  • 양시용 ((주)단바이오텍 생명과학연구소) ;
  • 박현주 (가톨릭대학교 생명공학) ;
  • 김형권 (가톨릭대학교 생명공학)
  • 발행 : 2008.12.28
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초록

사람 췌장 리파아제는 췌장에서 합성되어 소장으로 분비된 후, 음식물속의 지방을 가수분해하는 소화효소이다. 췌장 리파아제 단백질은 촉매활성도메인과 코리파아제 결합도메인의 2개의 도메인으로 구성되어 있다 본 연구에서는 리파아제 전체단백질과 촉매활성도메인 및 코리파아제 결함도메인 유전자를 PCR 방법으로 증폭하여 발현벡터에 넣은 후, 대장균에서 발현시켜 얻은 3가지 재조합 단백질을 산란계에 3차례 주사하여 면역반응을 유도하였다. 난황단백질을 분리하여 항체가를 측정한 결과, 코리파아제 결합도메인의 항원성이 가장 높은 것으로 밝혀졌다. 또한, 돼지 췌장 리파아제의 활성저해 실험에서도 코리파아제 결합도메인에 의해 만들어진 난황항체가 지방분해활성 저해작용이 가장 큰 것으로 밝혀졌다. 코리파아제 결합도메인 난황항체가 첨가된 지방을 실험동물에게 급여하고 혈중 주요 성분을 분석한 결과, 코리파아제 결합도메인에 의한 난황항체를 섭취한 경우에 혈중 중성지방의 수치가 대조군에 비해서 유의적으로 감소했음을 화인하였다. 결론적으로 코리파아제 결합도메인을 항원단백질로 사용하여 얻은 난황항체가 생체 내에서 췌장 리파아제의 활성을 억제할 수 있으며 비만억제제로의 개발 가능성이 있음을 보여주었다.

Human pancreatic lipase is a digestive enzyme which is synthesized in pancreas, secreted into small intestine, and there hydrolyze the fat in food. Pancreatic lipase protein composes of catalytic domain and colipase-binding domain. In this research, the gene segments corresponding to total protein, catalytic domain, and co lipase-binding domain were cloned by PCR method, inserted into an expression vector, and then used to transform Escherichia coli BL21 (DE3). The recombinant proteins produced were purified and injected intramuscularly three times into laying hens. The egg yolk antibodies (IgY) were obtained from the egg yolks and tested for their antibody titer. Among three IgY, the IgY against colipase-binding domain showed the highest antibody titer. All three IgY had inhibitory effects on the porcine pancreatic lipase. Among them, the IgY against colipase-binding domain showed the highest inhibition effects. The fat diet with corn oil and IgY was administrated to the experimental rats and their blood compositions were examined with time course. The triglyceride concentration of treated rats was decrease meaningfully when compared with those of control rats. This suggested that the IgY against colipase-binding domain antigen inhibited pancreatic lipase in vivo.

키워드

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