Korean Journal of Microbiology (미생물학회지)
- Volume 30 Issue 6
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- Pages.564-569
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- 1992
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- 0440-2413(pISSN)
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- 2383-9902(eISSN)
Purification and Some Properties of Glutamine Synthetase lsolated from Chlorobium limicola f. thiosulfatophilum NCIB 8327
Chlorobium limicola f. thiosulfatophilum NCIB 8327로부터 Glutamine Synthetase의 분리 및 특성분석
- Na, Jong-Uk (Department of Microbiology, College of Natural Sciences, Seoul National University) ;
- Kim, Ji-Yoon (Research Center for Molecular Microbiology, Seoul National University) ;
- Yoon, Hwan (Research Center for Molecular Microbiology, Seoul National University) ;
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Kang, Sa-Ouk
(Research Center for Molecular Microbiology, Seoul National University)
- 나종욱 (서울대학교 자연과학대학 미생물학과) ;
- 김지윤 (서울대학교 자연과학대학 미생물학과) ;
- 윤환 (서울대학교 분자미생물학연구센터) ;
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강사욱
(서울대학교 분자미생물학연구센터)
- Published : 1992.12.01
Abstract
A green sulfur bacterium, Chlorobium limicola f. thiosulfatophilum NCIB 8327, was grown in modified Pfennig's medium including glu1amate as a nitrogen source. Glutamine synthetase was isolated through a series of ultracentrifugation. DEAE-Sepharose CL-6B ion exchange chromatography. Sephacryl S-300 gel permeation chromatography, and preparative HPLC. The recovery and purification fold of the enzyme were 2% and 46.3. respectively. The isolated enzyme was homogeneous on UV-Visible spectrum and polyacrylamide gel electrophoretogram. The relative molecular mass of the native enzyme was estimated to be 280,000 by gel permeation chromatography. The enzyme consisted of ten subunits with relative similar molecular mass. 30.000. which was estimated by SDS-polyacrylamide gel electrophoresis. The optimal temperature and pH of the enzyme were
녹황색 세균인 Chlorobium limicola f. thiosulfatophilum NCIB 8327을 glutamate 가 질소원으로 포함된 변형 Pfennig 배지에서 배양한 다음 균체를 얻고, glutamine synthetase 효소를 초원심분리, DEAE-Sepharose CL-6B 이온교환 크로마토그래피, Sephacryl S-300 젤여과 크로마토그래피, 분취용 액체크로마토그래피의 과정을 통해 2% 수율에 46.3배의 정제배수로효소를 분리정제하였다. UV-VIS 흡수스펙트럼과 polycrylamide 젤 정기영동을 통해분리된 효소의 순수도는 확인하였다. Sephacryl S-3000 젤을 이용하여 얻은 효소의 분자량은 280 kDa 정도였으며, SDS-polyacrylamide 젤 전기영동 결과, 30 kDa 정도의 분자랴ㄹ을 갖는 단위체의 심합체로 추정된다. 이효소의 최적 온도와 pH 는 각각