摘要
用桔青霉菌株IM0 2 (PenicilliumcitrinumIM0 2 )发酵制得的核酸酶P1催化水解热变性DNA ,在研究温度、底物浓度、pH、酶加入量等因素对水解结果的影响的实验基础上,通过正交实验获得最佳的工艺条件:温度5 8℃,pH6 5 ,DNA质量浓度4 0g/L ,酶加入量4 % ,脱氧核苷酸的得率92 %。在此基础上推导出其催化米氏常数Km=5 818×10 -2 g/L。
Nuclease P1 obtained through fermentation by Penicillium citrinum was used for the enzymatic hydrolysis of the heat-denatured DNA, based on the investigation of the main effect factors, such as temperature, pH, the initial substrate concentration and the mass concentration of enzyme, the optimum hydrolytic con dition was obtained through orthogonal test, that was temperature 58 ℃, pH 6.5, the mass concentration of DNA 4.0 g/L, the concentration of enzyme 4% .The yield rate of DNA was 92%. Based on the calculation of experimental date, Michaelis constant was obtained as Km= 5. 818 × 10^-2 g/L.
出处
《生物加工过程》
CAS
CSCD
2005年第2期53-57,共5页
Chinese Journal of Bioprocess Engineering
基金
教育部跨世纪优秀人才培养基金
江苏省教育厅"3 3 3"工程基金项目
关键词
核酸酶P1
水解
热变性DNA
优化
米氏常数
nuclease P1
hydrolysis
heat-denatured DNA
optimization
Michaelis constant
