溶葡球菌酶前体及前体加工蛋白酶的生成控制和纯化

 为深入探讨溶葡球菌酶前体加工转化为成熟的溶葡球菌酶的机制,本文通过条件控制分别从Staphylococcus simulans的培养液中获得了溶葡球菌酶前体和它的加工蛋白酶,并分别通过HPLC和Affi-Gel 501亲和层析对它们进行了纯化,根据聚丙烯酰胺凝胶电泳和凝胶等电聚焦电泳,表明二者已基本上达到均一的程度。在此基础上,又进行了溶葡球菌酶前体的体外加工转化实验。

Biosynthesis Control and Purification of Prolysostaphin and its Processing Protease

Lysostaphin, an enzyme with molecular weight about 25000, secreted by Staphylococaus simulans, lyses all known Staphylococcus species. Lysostaphin gene codes for a proenzyme with molecular weight about 42000. The conversion of prolysostaphin is catalysed by processing protease. In order to get more prolysostaphin and its processing protease for inquiring into the mechanism of conversion of precursor, the present study was undertaken to isolate and to purify prolysostaphin and its processing protease. It was shown that both of them can be obtained in considerable quantity by controlling the preparation of inoculum, fermentational conditions and by choosing the right harvest timej and can be purified to homogeneity by HPLC and CM-Sephadex C-50, Affi-gel 501 chromatography, respectively. It was also shown that the processing protease can catalyze the conversion of prolysostaphin specifically without detectable degradation of lysostaphin.