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用脱氨再生几丁质亲和层析和羧甲基-纤维素CM52离子交换柱层析从芸苔(菜心)的茎叶中纯化了4种几丁质结合蛋白:CBPa、CBPb、CBPc和CBPd,SDS-PAGE显示CBPa和CBPb的分子量分别为29.04kDa和30.68kDa,凝胶过滤方法测定CBPa和CBPb的分子量分别为29.04kDa和31.2kDa,表明两者都是单体酶。4种CBP蛋白均有几丁质酶活性,其中CBPa和CBPb还有溶菌酶活性。EDTA对CBPa和CBPb溶菌酶的活性均无影响,各种类型的几丁质对CBPa和CBPb都有较强的吸附作用,PAS染色法分析表明CBPa和CBPb均为糖蛋白。
Four chitin binding proteins, CBPa, CBPb, CBPc, were purified from the stems and leaves of Brassica campestris L. with deamidation-regenerated chitin affinity chromatography and carboxymethyl-cellulose CM52 ion exchange column chromatography And CBPd. SDS-PAGE showed that the molecular weights of CBPa and CBPb were 29.04 kDa and 30.68 kDa, respectively. The molecular weights of CBPa and CBPb were 29.04 kDa and 31.2 kDa, respectively, by gel filtration assay, indicating that both were monomeric enzymes. All four CBP proteins had chitinase activity, of which CBPa and CBPb also had lysozyme activity. EDTA had no effect on the activity of CBPa and CBPb lysozyme, all kinds of chitin had strong adsorption on CBPa and CBPb, PAS staining analysis showed that both CBPa and CBPb were glycoproteins.