[1]刘嘉,贺淹才,施腾鑫,等.重组粘质沙雷氏菌几丁质酶C的纯化及酶学性质[J].华侨大学学报(自然科学版),2010,31(5):552-556.[doi:10.11830/ISSN.1000-5013.2010.05.0552]
　LIU Jia,HE Yan-cai,SHI Teng-xin,et al.Purification and Enzyme Properties of Recombinant Serratia marcescens Chitinase C[J].Journal of Huaqiao University(Natural Science),2010,31(5):552-556.[doi:10.11830/ISSN.1000-5013.2010.05.0552]
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重组粘质沙雷氏菌几丁质酶C的纯化及酶学性质()
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《华侨大学学报（自然科学版）》[ISSN:1000-5013/CN:35-1079/N]

卷:
第31卷
期数:
2010年第5期
页码:
552-556
栏目:
出版日期:
2010-09-20

文章信息/Info

Title:
Purification and Enzyme Properties of Recombinant Serratia marcescens Chitinase C
文章编号:
1000-5013(2010)05-0552-05
作者:
刘嘉; 贺淹才; 施腾鑫; 李梓君; 王珍珠
华侨大学化工学院
Author(s):
LIU Jia;  HE Yan-cai;  SHI Teng-xin;  LI Zi-jun;  WANG Zhen-zhu
College of Chemical Engineering, Huaqiao University, Quanzhou 362021, China
关键词:
粘质沙雷氏菌;  重组;  几丁质酶C;  分离纯化;  酶学性质
Keywords:
Serratia marcescens;  recombinant;  chitinase C;  isolation and purification;  enzyme properties
分类号:
Q78
DOI:
10.11830/ISSN.1000-5013.2010.05.0552
文献标志码:
A
摘要:
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4 h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.
Abstract:
To obtain high pure chitinase C from constructed recombinant E.coli pET-22b-chiC,we decreased culture temperature to increase soluble expression of Serratia marcescens chitinase C.The expressed product was purified to electrophoretic homogeneity by immobilized metal ion affinity chromatography(IMAC)and Phenyl-Sepharose hydrophobic interaction chromatography(HIC).The enzyme properties research indicated that the purified chitinase C was a monomeric protein with a molecular weight of 51.8 ku.The optimum pH was 5.0 and the optimum temperature was 55 ℃.And its activity still kept above 90% after 1 h at 55 ℃.Results also showed that the activity of chitinase was significantly inhibited by Cu2+,Hg2+,Co2+,Mg2+,while promoted by Fe2+,Zn2+,Sn2+,Ba2+ and also significantly promoted by Mn2+.

参考文献/References:

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备注/Memo

备注/Memo:
福建省自然科学基金资助项目(C04010011)
更新日期/Last Update: 2014-03-23