红球菌菌株腈水合酶提取方法优化  

Optimization methods of extraction of the nitrile hydratase from Rhodoccus sp. Tccc28001

作　　者：王世伟[1] 王卿惠[1] 翟丽萍[1] 刘军[1] 闫冀川 李小鹏[1] 

机构地区：[1]齐齐哈尔大学生命科学与农林学院,黑龙江齐齐哈尔161006

出　　处：《高师理科学刊》2016年第8期47-50,共4页Journal of Science of Teachers＇College and University

基　　金：黑龙江省教育厅科学技术研究项目(12541855)

摘　　要：研究了红球菌（Rhodoccus sp.Tccc28001）腈水合酶初步提取实验.对腈水合酶提取的菌体培养时间、细胞的超声破碎方法和硫酸铵分级分离方法进行了优化.结果表明,提取的最佳条件为：培养96 h收获菌体;在4℃下溶菌酶处理菌悬液40 h后进行超声波处理,400 W超声破碎60min,分2次进行,每次30 min（60个循环,15 s超声,15 s间歇）;然后使用40 mmol/L磷酸缓冲液,经20%~70%饱和度硫酸铵沉淀,能获得最高产率的腈水合酶.此研究为腈水合酶的层析纯化奠定了基础.The extraction of NHase from Rhodoccus sp.Tccc28001 has been optimised using different methods such as cell culture time,cell ultrasonic fragmentation,ammonium sulfate fractionation.The results showed that the optimum conditions of the NHase extraction ware as follows,the bacteria fermentation culture was harvested for 96 h.After the bacteria were hydrolyzed by lysozyme for 40 h（at 4 ℃）,the ultrasonic fragmentation was carried out at 400 W for 60 min and this segment was divided into two steps,each step needs 30 min（60 cycles,15 s ultrasonic,15 s interval）,40 mmol/L phosphate buffers was used in 20%~70% saturation ammonium sulfate precipitation for the maximum yield of the nitrile hydratase.

关 键 词：红球菌 腈水合酶 优化 

分 类 号：Q814.1[生物学—生物工程]