地衣芽孢杆菌弹性蛋白酶纯化和性质研究  

Purification and Characteristics of Elastase Produced by Bacillus licheniformis

作　　者：傅明亮[1] 刘晓杰[1] 刘婧[1] 何国庆[1] 陈启和[1] 

机构地区：[1]浙江大学生物系统工程与食品科学学院,浙江杭州310029

出　　处：《食品科学》2011年第7期216-219,共4页Food Science

基　　金：浙江省自然科学基金项目(Y307405)

摘　　要：用地衣芽孢杆菌发酵液制备弹性蛋白酶粗酶液,采用硫酸铵分级沉淀和Sephadex凝胶柱层析的方法分离纯化弹性蛋白酶,并对弹性蛋白酶的酶学性质进行研究。结果表明:弹性蛋白酶粗酶液经40%～70%饱和度的硫酸铵纯化后比活力提高到120U/mg,经凝胶柱层析纯化后比活力可达到292U/mg,纯化倍数为12.6,SDS-PAGE法证实弹性蛋白酶分子质量为29.5kD。对酶学性质的研究结果表明:弹性蛋白酶最适反应温度为55℃,最适反应pH值为7.4,以刚果红-弹性蛋白为底物,米氏常数Km为9.67mg/mL。低浓度金属离子Ca2+和K+对酶活力有促进作用,而Mg2+、Mn2+、Zn2+和Al3+对酶活力则有抑制作用。The fermentation supernatant（rich in elastase） of Bacillus licheniformis ZJUEL31410,a strain isolated from soil,was used as a crude enzyme solution,from which elastase was separated by（NH4）2SO4 segmentation precipitation and purified by Sephadex gel column chromatography.The enzymatic characteristics of purified elastase were measured.The specific activity of elastase could be increased to 120 U/mg through（NH4）2SO4 precipitation with 40%-70% saturation degree and continually increased to 292 U/mg after Sephadex gel column purification and the purification fold was 12.6.SDS-PAGE studies demonstrated that the enzyme had a molecular weight of 29.5 kD.The optimum reaction temperature and pH were 55 ℃ and 7.4,respectively,and the Michaelis constant Km was 9.67 mg/mL with elastin-Congo red as the substrate.Low concentrations of Ca2＋and K＋ could enhance elastase activity,while Mg2 ＋,Mn2 ＋,Zn2 ＋and Al3 ＋ revealed an inhibition effect.

关 键 词：地衣芽孢杆菌ZJUEL31410 弹性蛋白酶 纯化 酶学性质 

分 类 号：TS201[轻工技术与工程—食品科学]