생명과학회지 (Journal of Life Science)

  • 제18권6호
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  • Pages.845-851
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  • 2008
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
권호 표지
DOI QR코드

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(β-lactam계 항생물질 저항성을 지닌 Bacillus sp. J105 균주로부터 분비되는 베타 락탐 분해효소의 정제 및 특성

Purification and Characterization of β-Lactamase Secreted from Bacillus sp. J105 Strain having β-Lectam Antibiotics Resistance.

  • 조경순 (부산광역시 보건환경연구원) ;
  • 강병원 (동아대학교 BK21 실버바이오 사업단) ;
  • 서민정 (동아대학교 생명공학과) ;
  • 이영춘 (동아대학교 생명공학과) ;
  • 이재헌 (동아대학교 유전공학과) ;
  • 주우홍 (창원대학교 생물학과) ;
  • 최영현 (동의대학교 한의학대학 생화학교실) ;
  • 임학섭 ((주)천년약속 바이오연구소) ;
  • 김정인 (인제대학교 식품생명과학부) ;
  • 서권일 (순천대학교 식품영양학과) ;
  • 정영기 (동아대학교 생명공학과, (주)천년약속 바이오연구소)
  • Cho, Kyeong-Soon (Public Health and Environment Institute of Pusan) ;
  • Kang, Byoung-Won (BK21 Center for Silver-Bio industrialization Project, Dong-A University) ;
  • Seo, Min-Jeong (Department of Biotechnology, Dong-A University) ;
  • Lee, Young-Choon (Department of Biotechnology, Dong-A University) ;
  • Lee, Jai-Heon (Department of Genetic Engineering, Dong-A University) ;
  • Joo, Woo-Hong (Department of Biology, Changwon National University) ;
  • Choi, Yung-Hyun (Department of Biochemistry, College of Oriental Medicine, Dong-eui University) ;
  • Lim, Hak-Seob (Department Bioinstitute, MILLENNIUM PROMISE CO., LTD.) ;
  • Kim, Jeong-In (School of Food and Life Science, Inje University) ;
  • Seo, Kwon-Il (Department of Food and Nutrition, Sunchon National University) ;
  • Jeong, Yong-Kee (Department of Biotechnology, Dong-A University, Department Bioinstitute, MILLENNIUM PROMISE CO., LTD.)
  • 발행 : 2008.06.30
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초록

Bacillus sp. J105 strain으로부터 유도된 ${\beta}-lactamase$는 ammonium sulfate 침전, 이온 교환 칼럼 크로마토그래피, 겔 여과 등의 과정을 거쳐 SDS-PAGE에서 단일 band로 정제하였다. 정제된 효소의 분자량은 31 kDa이었으며 등전점은 7.35이었다. 효소반응의 최적 pH와 온도는 각각 5와 $40^{\circ}C$이었다. 정제 단백질의 총 아미노산 조성의 분석 결과, Gly과 Ala이 각각 14.1과 13.3 mole%로 가장 많은 아미노산 잔기를 차지하고 있었다. 정제 효소의 ampicillin을 기질로 하였을 때의 Km값은 1.33 mM이었고 Vmax값은 0.36 mM/ml이었다.

${\beta}-Lactamase$, secreted from Bacillus sp. J105 strain was purified to a single band on SDS-PAGE by ammonium sulfate precipitation, ion exchange column chromatography and gel-filtration. The molecular weight of the purified enzyme was 31 kDa on SDS-PAGE and its isoelectric point was 7.35. Optimal pH and temperature for enzymatic reaction were 5 and $40^{\circ}C$, respectively. As a result of total amino acid composition analysis of the purified enzyme, Gly and Ala were occupied 14.1 and 13.3 mole %, respectively. Km and Vmax value of purified enzyme were 1.33 mM and 0.36 mM/ml using ampicillin as a substrate, respectively.

키워드

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