海洋微生物溶菌酶的纯化与性质研究  

Purification and Characterization of a Lysozyme from a Marine Microorganism

作　　者：邹艳丽[1] 孙谧[2] 王跃军[2] 

机构地区：[1]上海水产大学食品学院,上海200090 [2]中国水产科学院黄海水产研究所海洋酶与酶工程实验室,青岛266071

出　　处：《生物工程学报》2005年第3期420-424,共5页Chinese Journal of Biotechnology

基　　金：国家高技术研究发展计划资助项目 (No .2 0 0 3AA62 5 0 70 )~~

摘　　要：海洋微生物溶菌酶发酵上清液经超滤、CM_SepharoseFF阳离子交换层析和SephadexG_10 0凝胶过滤层析纯化得到电泳纯的溶菌酶,纯化倍数为34 7,活力回收为2 4 1%。对纯化溶菌酶性质研究表明,该酶分子量约为39kD ,对溶壁微球菌的最适作用温度为35℃,最适作用pH为8 0 ,在5 0℃以下和pH 5 0～10 0之间都有较好的稳定性,与常见金属离子和化学试剂有良好的配伍性,广谱杀菌,对多种致病菌也有较强的溶菌作用。A novel lysozyme was purified from a marine microorganism and its major characteristics were studied. Cell-free supernatant was prepared by centrifugation of culture broth, ultrafiltration using a hollow fiber (molecular weight cut off, 50kD) and concentration using a hollow fiber (molecular weight cut off, 10kD). The crude lysozyme was purified 34.7 fold to electrophoretic homogeneity with a recovery of 24.1% by CM-Sepharose FF cationic-exchange and Sephadex G-100 gel chromatography. The relative molecular weight of this lysozyme was determined as about 39 kD. The optimum pH and temperature towards Micrococcus lysodleikticus were pH 8.0 and 35℃ respectively, and the enzyme was stable at temperature below 50℃ and pH 5.0～10.0. The lysozyme activity was slightly enhanced by Zn 2+ and Cu 2+ and slightly inhibited by Mn 2+ and Ag + . The lysozyme showed good compatibility to many common chemical agents such as EDTA (0.1%) and KH 2 PO 4 (1.0%). The lysozyme had broad-spectrum against many bacteria, including a number of pathogens, which were resistant to egg-white lysozyme.

关 键 词：海洋微生物 溶菌酶 分离纯化 理化性质 

分 类 号：Q936[生物学—微生物学]