全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

重组粘质沙雷氏菌几丁质酶C的纯化及酶学性质

DOI: 10.11830/ISSN.1000-5013.2010.05.0552, PP. 552-556

Keywords: 粘质沙雷氏菌,重组,几丁质酶C,分离纯化,酶学性质

Full-Text   Cite this paper   Add to My Lib

Abstract:

为从已构建的重组大肠杆菌pET-22b-chiC获得高纯的几丁质酶C,通过降低培养温度,提高粘质沙雷氏菌几丁质酶C的可溶性表达.表达产物经镍柱亲和层析(IMAC)和Phenyl-Sepharose疏水层析(HIC)分离纯化后,得到电泳纯的几丁质酶.酶学性质研究表明,纯化的几丁质酶C为单体蛋白,相对分子质量为51.8ku;最适pH值为5.0,最适温度为55℃,在55℃的条件下保温4h后仍有90%以上的酶活力.研究结果还表明,Cu2+,Hg2+,Co2+,Mg2+对酶活力均有明显抑制作用,而Fe2+,Zn2+,Sn2+,Ba2+对酶活力有一定促进作用,Mn2+对酶活力明显促进作用.

References

[1]  SUZUKI K, SUGAWARA N, SUZUKI M. Chitinase A, B, and C1 of Serratia marcescens 2170 produced by recombinant Escherichia coli:Enzymatic properties and synergism on chitin degradation [J]. Bioscience, Biotechnology, and Biochemistry, 2002(5):1075-1083.
[2]  BRURBERG M, SYNSTAD B, KLEMSDAL S S. Chitinases from Serratia marcescens [EB/OL]. http://davapc1.bioch.dundee.ac.uk/pdf/chirev.pdf, 2000.
[3]  GAL S W, CHOI Y J KIM Y C. Cloning of the 52-kDa chitinase gene from Serratia marcescens KCTC2172 and its proteolytic cleavage into an active 35-kDa enzyme [J]. FEMS Microbiology Letters, 1998(1):151-158.
[4]  贺淹才, 刘爱花, 张荣奎. 嗜麦芽窄食单胞菌产生的几丁质酶的特性 [J]. 华侨大学学报(自然科学版), 2008(2):245-249.
[5]  魏巍, 贺淹才, 方柏山. 粘质沙雷氏菌几丁质酶(ChiC)基因克隆及其生物信息学分析 [J]. 江西农业大学学报, 2006(3):444-448.doi:10.3969/j.issn.1000-2286.2006.03.027.
[6]  SUZUKI K, TAIYOJI M, SUGAWARA N. The third chitinase gene (chiC) of Serratia marcescens 2170 and the relationship of its product to other bacterial chitinases [J]. Biochemical Journal, 1999(3):587-596.doi:10.1042/0264-6021:3430587.
[7]  萨姆布鲁克?J, 拉塞尔 D W, 黄培堂. 分子克隆实验指南 [M]. 北京:科学出版社, 2002.
[8]  SYNSTAD B, VAAJE-KOLSTAD G, CEDERKVIST F. Expression and characterization of endochitinase C from Serratia marcescens BJL200 and its purification by a one-step general chitinase purification method [J]. Bioscience, Biotechnology, and Biochemistry, 2008(3):715-723.doi:10.1271/bbb.70594.
[9]  TSAFFRIR Z, ZVI S. Linearization of the bradford protein assay increases its sensitivity:Theoretical and experimental studies [J]. Analytical Biochemistry, 1996(2):302-308.
[10]  唐亚雄, 赵建, 丁诗华. 产气肠杆菌几丁质酶的分离纯化及性质研究 [J]. 微生物学报, 2001(1):82-86.doi:10.3321/j.issn:0001-6209.2001.01.015.
[11]  SYNSTAD B, GASEIDNES S, AALTEN D M F. Mutational and computational analysis of the role of conserved residues in the active site of a family 18 chitinase [J]. European Journal of Biochemistry, 2004(2):253-262.doi:10.1046/j.1432-1033.2003.03923.x.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133