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[一种海洋微生物溶菌酶的纯化与特性分析]

[Purification and characterization of a lysozyme from a marine microorganism].

作者信息

Zou Yan-Li, Sun Mi, Wang Yue-Jun

机构信息

Laboratory of Marine Enzyme and Enzyme Engineering, Yellow Sea Fisheries Research Institute, Qingdao 266071, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2005 May;21(3):420-4.

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 (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 degrees C respectively, and the enzyme was stable at temperature below 50 degrees C and pH 5.0 - 10.0. 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 (0.1%) and KH2 PO4 (1.0%). The lysozyme had broad-spectrum against many bacteria, including a number of pathogens, which were resistant to egg-white lysozyme.

摘要

从一种海洋微生物中纯化出一种新型溶菌酶,并对其主要特性进行了研究。通过对培养液进行离心、使用中空纤维(截留分子量50kD)超滤以及使用中空纤维(截留分子量10kD)浓缩来制备无细胞上清液。粗溶菌酶通过CM-Sepharose FF阳离子交换和Sephadex G-100凝胶色谱纯化至电泳纯,纯化倍数为34.7倍,回收率为24.1%。该溶菌酶的相对分子量测定约为39kD。其对溶壁微球菌的最适pH和温度分别为pH 8.0和35℃,该酶在50℃以下及pH 5.0 - 10.0时稳定。Zn2+和Cu2+对溶菌酶活性有轻微增强作用,而Mn2+和Ag+有轻微抑制作用。该溶菌酶对许多常见化学试剂如EDTA(0.1%)和KH2PO4(1.0%)具有良好的兼容性。该溶菌酶对许多细菌具有广谱抗菌活性,包括一些对蛋清溶菌酶耐药的病原体。

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