重组粘质沙雷氏菌几丁质酶C的纯化及酶学性质  

Purification and Enzyme Properties of Recombinant Serratia marcescens Chitinase C

作　　者：刘嘉[1] 贺淹才[1] 施腾鑫[1] 李梓君[1] 王珍珠[1] 

机构地区：[1]华侨大学化工学院,福建泉州362021

出　　处：《华侨大学学报（自然科学版）》2010年第5期552-556,共5页Journal of Huaqiao University(Natural Science)

基　　金：福建省自然科学基金资助项目(C04010011)

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.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＋.

关 键 词：粘质沙雷氏菌 重组 几丁质酶C 分离纯化 酶学性质 

分 类 号：TQ920.6[轻工技术与工程—发酵工程] TQ929.2