Purification of Alkaline Restriction Endonuclease from Alkalophilic Bacillus sp. 8-13

제한효소 생성능을 지닌 알칼리성 Bacillus sp. 8-13 균주로부터 알칼리성 제한효소의 정제

  • Bae, Moo (Department of Biology, College of Pharmarcy, Ewha Womans University) ;
  • Lee, Jee-Eun (Department of Biology, College of Pharmarcy, Ewha Womans University) ;
  • Park, Kyoung-Sook (Department of Biology, College of Pharmarcy, Ewha Womans University) ;
  • Lee, Kang-Man (Department of Industrial Pharmarcy, Ewha Womans University)
  • 배무 (이화여자대학교 자연과학대학 생물학과) ;
  • 이지은 (이화여자대학교 자연과학대학 생물학과) ;
  • 박경숙 (이화여자대학교 자연과학대학 생물학과) ;
  • 이강만 (이화여자대학교 약학대학 제약학과)
  • Published : 1992.06.01

Abstract

Twenty-four bacterial strains among alkalophillic bacteria isolated from soil samples were examined for the presence of type II restriction endonuclease in aerobic culture. One strain was found to contain specific enzyme to cleave lambda DNA. The characteristics of this microorganism is the ability to grow well in alkalophilic and high temperature condition, that is at pH 10.3 and $50^{\circ}C$. This strain was tentatively identified to Bacillus alkaloPhilus subsp. halodurans when morphological, physiological and biochemical characteristics were examined. The enzyme was purified from crude extract by streptomycin sulfate, ammonium sulfate precipitation, which was followed by DEAE-cellulose and phosphocellulose ion exchange column chromatography, and the subunit molecular weight was about, 32,000 daltons by polyacrylamide gel electrophoresis containing 0.1% SDS.