摘要
以菜用大豆为原料,直投式纳豆芽孢杆菌作为发酵剂,纳豆激酶活力作为评价指标,通过单因素和正交试验优化纳豆激酶发酵工艺条件;通过盐溶、硫酸铵分段盐析、二乙氨乙基(DEAE)阴离子交换柱分离纯化纳豆激酶,并采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)确定其分子质量,并进行酶学性质和体外溶栓效果分析。结果表明,菜用大豆发酵生产纳豆激酶的最佳条件为:每100 g蒸煮菜用大豆接入1.15×10^11 CFU/g直投发酵剂,发酵温度37℃、发酵时间36 h、后熟时间18 h时,此时纳豆激酶活力可达2326.60 IU/g。纳豆激酶的分子质量介于25~35 kDa之间,最适作用温度、pH值分别为37℃、8.0,当温度低于40℃、pH值在6.0~8.0时较稳定;体外溶栓实验表明纳豆激酶具有良好的体外溶栓效果。
Using vegetable soybean as the substrate,Directed-Vat-Set Bacillus natto as the starter,and nattokinase activity as evaluation index,the fermentation conditions of nattokinase were optimized by single-factor experiments and orthogonal experiments.Then the extracted nattokinase was isolated and purified by salting-in,salting-out with gradient ammonium sulfate solution and dicthylaminoethyl(DEAE)anion exchange column,and the molecular mass was determined through sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE).Finally,the enzymatic property and in vitro thrombolytic properties were also analyzed.The results shown that the optimal conditions for nattokinase production through vegetable soybean fermentation were obtined as follows:On 100 g vegetable soybean base,1.15×10^11 CFU/g Bacillus natto was inoculated as starter with fermentation temperature 37℃,fermentation time 36 h and post-ripening time 18 h.The purified nattokinase activity was up to 2236.60 IU/g with molecular mass between 25-35 kDa.The optimal temperature and pH for nattokinase activity was 37℃and 8.0,respectively.It showed good stability under 40℃and pH 6.0-8.0.The results of in vitro thrombolysis showed that nattokinase had good thrombolysis effect.
作者
李秀凉
李倩文
朱玉
韩晓云
孙庆申
宋永
LI Xiuliang;LI Qianwen;ZHU Yu;HAN Xiaoyun;SUN Qingshen;SONG Yong(Engineering Research Center of Agricultural Microbiology Technology,Ministry of Education,Heilongjiang University,Harbin 150500,China;Key Laboratory of Molecular Biology,College of Heilongjiang Province,School of Life Sciences,Heilongjiang University,Harbin 150080,China;Key Laboratory of Microbiology,College of Heilongjiang Province,School of Life Sciences,Heilongjiang University,Harbin 150080,China)
出处
《中国酿造》
CAS
北大核心
2020年第12期106-112,共7页
China Brewing
关键词
纳豆激酶
发酵
分离纯化
稳定性
体外溶栓
nattokinase
fermentation
separation and purification
stability
in vitro thrombolysis