본 연구는 Streptomyces subrutilus P5의 천연 Fe superoxide dismutase (FeSOD)와 유전자 재조합 기술로 생산된 6xHis-태그가 결합된 Fe superoxide dismutase (6xHis- FeSOD)의 활성을 비교하여 6xHis-태그의 효소에 대한 ...
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https://www.riss.kr/link?id=A102310260
박중호 (단국대학교) ; 김재헌 (단국대학교) ; Park, Joong-ho ; Kim, Jae-heon
2016
Korean
KCI등재,SCOPUS
학술저널
230-235(6쪽)
0
0
상세조회0
다운로드국문 초록 (Abstract)
본 연구는 Streptomyces subrutilus P5의 천연 Fe superoxide dismutase (FeSOD)와 유전자 재조합 기술로 생산된 6xHis-태그가 결합된 Fe superoxide dismutase (6xHis- FeSOD)의 활성을 비교하여 6xHis-태그의 효소에 대한 ...
본 연구는 Streptomyces subrutilus P5의 천연 Fe superoxide dismutase (FeSOD)와 유전자 재조합 기술로 생산된 6xHis-태그가 결합된 Fe superoxide dismutase (6xHis- FeSOD)의 활성을 비교하여 6xHis-태그의 효소에 대한 영향을 알아보기 위하여 수행되었다. 두 효소 모두 최적 pH는 7로 동일하였으나 6xHis-태그에 의해서 pH 범위는 축소되었다. 천연 효소는 pH 4-9의 범위에서 안정성을 보인 반면 6xHis-FeSOD는 pH 9에서 안정성이 상실되었다. 두 효소의 최적 온도는 차이가 없으나 열 안정성에 있어서는 천연 효소는 $40^{\circ}C$ 이하에서 720분까지 안정성을 유지하였으나 6xHis-FeSOD는 $20^{\circ}C$에서도 360분 이내에 활성을 잃는 것으로 나타났다. $H_2O_2$의 6xHis-FeSOD에 대한 저해는 0.5 mM에서 나타났다. 따라서 6xHis-FeSOD는 효소활성은 유지되더라도 열 안정성이 크게 감소되는 결과를 얻었다. 이것은 6xHis-태그가 활성부위 보다는 단백질 전체 구조에 더 많은 영향을 미친 결과라고 생각되었다.
다국어 초록 (Multilingual Abstract)
This study was carried out to analyze the differences in enzyme activity and stability between the native Fe superoxide dismutase (FeSOD) and the 6xHis-tagged superoxide dismutase (6xHis-FeSOD) of Streptomyces subrutilus P5. The optimum pHs for both n...
This study was carried out to analyze the differences in enzyme activity and stability between the native Fe superoxide dismutase (FeSOD) and the 6xHis-tagged superoxide dismutase (6xHis-FeSOD) of Streptomyces subrutilus P5. The optimum pHs for both native FeSOD and 6xHis-FeSOD were 7, while the pH range of the activity was narrower for the 6xHis-FeSOD. The native FeSOD was stable at pH 4-9, but the 6xHis-FeSOD lost its stability at pH > 9. The temperatures of the optimum activities were same for both types of enzymes. However, the heat stability of the 6xHis-FeSOD was clearly decreased; even at $20^{\circ}C$ the enzyme lost the activity after 360 min. In contrast, the native FeSOD was stable after 720 min at below $40^{\circ}C$. $H_2O_2$ inhibition was occurred already at 0.5 mM for the 6xHis-tagged enzyme. Therefore, from the results that the 6xHis-FeSOD retained the enzyme activity at pH 6-7 and $20-40^{\circ}C$, it can be assumed that the protein structure became destabilized under different storage conditions and sensitive to the enzyme inhibitor.
참고문헌 (Reference)
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6 Zhao, G., "Structures of the N-acetyltransferase domain of Xylella fastidiosa N-acetyl-L-glutamate synthase/kinase with and without a His tag bound to N-acetyl-L-glutamate. Acta Crystallogr" 71 : 86-95, 2015
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1 Sato, S., "pH-dependent stability and folding kinetics of a protein with an unusual alpha-beta topology: the C-terminal domain of the ribosomal protein L9" 318 : 571-582, 2002
2 Sayari, A., "The N-terminal His-tag affects the enantioselectivity of staphylococcal lipases: A monolayer study" 313 : 261-267, 2007
3 Miller, A. F, "Superoxide dismutases: active sites that save, but a protein that kills" 8 : 162-168, 2004
4 Beauchamp, C., "Superoxide dismutase: improved assays and an assay applicable to acrylamide gels" 44 : 276-287, 1971
5 Lin, C. T., "Subunit interaction enhances enzyme activity and stability of sweet potato cytosolic Cu/Zn-SOD purified by a His-tagged recombinant protein method" 28 : 303-311, 1995
6 Zhao, G., "Structures of the N-acetyltransferase domain of Xylella fastidiosa N-acetyl-L-glutamate synthase/kinase with and without a His tag bound to N-acetyl-L-glutamate. Acta Crystallogr" 71 : 86-95, 2015
7 Van Gelder, P., "Structural and functional characterization of a His-tagged PhoE pore protein of Escherichia coli" 229 : 869-875, 1996
8 Pitcher, D. G., "Rapid extraction of bacterial genomic DNA with guanidium thiocyanate" 8 : 151-156, 1989
9 Desai, K., "Purification and biochemical characterization of a SOD from the soluble fraction of the cyanobacterium, Spirulina platensis" 23 : 1661-1666, 2007
10 Farr, S.B., "Oxidative stress in Escherichia coli and Salmonella typhimurium" 55 : 561-568, 1991
11 김재헌, "Iron Containing Superoxide Dismutase of Streptomyces subrutilus P5 Increases Bacterial Heavy Metal Resistance by Sequestration" 한국미생물학회 50 (50): 179-184, 2014
12 Lupi, A., "Human recombinant prolidase from eukaryotic and prokaryotic sources" 273 : 5466-5478, 2006
13 Khan, F., "Histidine affinity tags affect MSP1(42)structural stability and immunodominance in mice" 7 : 133-147, 2012
14 Carson, M., "His-tag impact on structure. Acta Crystallogr" 63 : 295-301, 2007
15 Klose, J., "Hexahistidine tag position influences disulfide structure but not binding behavior of in vitro folded N-terminal domain of rat corticotropin-releasing factor receptor type 2a" 13 : 2470-2475, 2004
16 Halliwell, B., "Free Radicals in Biology and Medicine" Claredon Press 299-365, 1999
17 Panek, A., "Effects of the polyhistidine tag on kinetics and other properties of trehalose synthase from Deinococcus geothermalis" 60 : 163-166, 2013
18 Ames, B.N., "DNA and Free Radicals" Ellis-Horwlld 1-15, 1993
19 Chant, A., "Attachment of a histidine tag to the minimal zinc finger protein of the Aspergillus nidulans gene regulatory protein AreA causes a conformational change at the DNA-binding site" 39 : 152-159, 2005
20 Horng, J. C., "Analysis of the pH-dependent and stability of histidine point mutants allows characterization of the denatured state and transition state for protein folding" 345 : 163-173, 2004
21 So, N. W., "A lead-absorbing protein with SOD activity from Streptomyces subrutilus" 194 : 93-98, 2001
한반도 주변 해역으로부터 혐기성 미생물의 분리 및 분리 미생물의 특성 분석
새로운 Psychrobacter sp. ArcL13 유래 저온활성 지질분해효소 : 유전자 분리동정, 대장균에서의 발현, refolding 및 특성 연구
학술지 이력
연월일 | 이력구분 | 이력상세 | 등재구분 |
---|---|---|---|
2023 | 평가예정 | 해외DB학술지평가 신청대상 (해외등재 학술지 평가) | |
2020-01-01 | 평가 | 등재학술지 유지 (해외등재 학술지 평가) | |
2013-12-02 | 학술지명변경 | 외국어명 : The Korean Journal of Microbiology -> Korean Journal of Microbiology | |
2010-01-01 | 평가 | 등재학술지 유지 (등재유지) | |
2008-01-01 | 평가 | 등재학술지 유지 (등재유지) | |
2006-01-01 | 평가 | 등재학술지 유지 (등재유지) | |
2004-01-01 | 평가 | 등재학술지 유지 (등재유지) | |
2001-01-01 | 평가 | 등재학술지 선정 (등재후보2차) | |
1998-07-01 | 평가 | 등재후보학술지 선정 (신규평가) |
학술지 인용정보
기준연도 | WOS-KCI 통합IF(2년) | KCIF(2년) | KCIF(3년) |
---|---|---|---|
2016 | 0.21 | 0.21 | 0.21 |
KCIF(4년) | KCIF(5년) | 중심성지수(3년) | 즉시성지수 |
0.26 | 0.24 | 0.48 | 0.02 |