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《Journal of Zhejiang University(Engineering Science)》 2004-10
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Screening of Bacillus subtilis natto and optimization of nattokinase fermentation

MEI Le-he~1, HU Shen~1, XU Jing~2, YAO Shan-jing~1, ZHENG Xiao-dong~1(1. Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou 310027, China; 2. Hangzhou Eastchina Pharmaceutical Company, Hangzhou 310011, China)  
Nattokinase, which is extracted from traditional Japanese food Natto is a fibrinolytic enzyme with strong and specific thrombolytic activity. A wild type Bacillus subtilis natto strain was screened from traditional Japanese food Natto with an improved medium-fibrinogen plate. The screening strategy included casein plate selecting and shaking culturing. The highest production of Nattokinase was 970IU/mL. The fermentation condition of nattokinase production by Bacillus subtilis natto was optimized with response surface methodology (RSM). In the fist optimization step, Plackett-Burman design was used to evaluate the influence of related factors. Tryptone influenced nattokinase production positively while Na_2HPO_4, temperature and age of inoculum influenced production negatively. The others had no significant influence on nattokinase production. The path of steepest ascent was used to approach the optimal region of the fermentation condition subsequently. The concentrations of tryptone and Na_2HPO_4 , and the age of inoculum were further optimized using central composite designs and response surface analysis. The optimized condition allowed nattokinase production to increase from 1005IU/mL to 1314IU/mL.
【Fund】: 国家自然科学基金资助项目(20176050);; 浙江省科技计划资助项目(2004C33036);; 浙江省回国人员基金资助项目;; 教育部回国人员基金资助项目;; 中德合作DAADCSC科技人员交流项目(PPP).
【CateGory Index】: TQ920.4
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