【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.更多还原

【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+.更多还原