海洋微生物溶菌酶的纯化与性质研究.pdfVIP

21 3 Vol.21 No. 3 2005 5 ChineseJournal of Biotechnology May 2005 Purification and Characterization of a Lysozyme from a Marine Microorganism 2 1* 1 邹艳丽, 孙 谧 , 王跃军 2 1* 1 ZOU Yan_Li , SUN Mi and WANG Yue_Jun 1 , 266071 2 , 200090 1Laboratory of Marine Enzyme and Enzyme Engineering, Yellow Sea Fisheries Research Institute, ingdao, 266071, China 2 College of Food Science, ShanghaiFisheries University , Shanghai 200090, China 海洋微生物溶菌酶发酵上清液经超滤CM_Sepharose 阳离子交换层析和Sephadex G_100 凝胶过滤层析纯化得到 泳纯的溶菌酶, 纯化倍数为347, 活力回收为241% 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80,在50 以下和 pH 50~ 100 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 海洋微生物, 溶菌酶, 分离纯化, 理化性质 Q55 A 2005) Abstract 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 ( molecularweight cut off, 50kD) and concentration using a hollow fiber ( molecular weight cut off, 10kD) . The crude lysozyme was purified 347 fold to electrophoretic homogeneity with a recovery of 241% by CM_Sepharose 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 temperaturetowards Micrococcus lysodleikticus were pH 80 and 35 respectively, and the enzyme was stable at temperature below 50 and pH 50~ 100. The lysozyme activity was slightly enhanced by Zn2+ and Cu2+ and slightly inhibited by Mn2+ + and Ag . The lysozyme showed good compatibility to many common chemical agents such as EDTA ( 01%) and KH PO