纳豆激酶发酵条件的优化.pdfVIP

※生物工程 食品科学 2009, Vol. 30, No. 01 151 董 超，李 楠，程辉彩，张根伟，史延茂 (河北省生物研究所，河北石家庄 050081) ：利用Plackett-Burman 方法对影响纳豆芽孢杆菌液体发酵的各因素进行评价，筛选出对产纳豆激酶有显著效 应的主要因素是豆饼粉浓度。在此基础上，用响应面分析法对发酵培养基中的豆饼粉浓度、CaCl2 浓度、葡萄糖 浓度进行优化。所得的最优培养基成分为葡萄糖 1% 、豆饼粉 3% 、KH2 PO4 0.2% 、K2 HPO4 ·3H2 O 0.26% 、 MgSO4 ·7H2O 0.1% 、CaCl2 0.12% 。拟合实验结果显示，纳豆激酶液体发酵液的酶活由初始培养条件的约1800U/ml 提高到优化后的2226.06U/ml 。 ：纳豆激酶；纳豆芽孢杆菌；Plackett-Burman 设计；响应面分析法 Optimization of Fermentation Conditions for Production of Nattokinase DONG Chao ，LI Nan ，CHENG Hui-cai ，ZHANG Gen-wei ，SHI Yan-mao (Hebei Institute of Biology, Shijiazhuang 050081, China) Abstract ：This study aimed to optimize the fermentation conditions for nattokinase production by Bacillus subtilis Natto with response surface methodology. Firstly, Plackett-Burman design was used to evaluate the effects of related factors on nattokinase production. It was found that soybean powder concentration is the most signficant positive factor, followed by concentrations of glucose and CaCl2 (negative factor). And then, the concentrations of soybean powder, glucose and CaCl2 were further optimized using response surface analysis. Results revealed that the optimal medium constituents for liquid fermentation are as follows: glucose 1%, soybean powder 3%, KH2PO4 0.2%, K2HPO4 ·3H2O 0.26%, MgSO4 ·7H2O 0.1% and CaCl2 0.12%. The optimized conditions allow nattokinase production to increase from 1800 U/ml to 2226.06 U/ml. Key words ：nattokinase ；Bacillus subtilis Natto ；Plackett-Burman design ；response surface methodology 中图分类号：Q946.5 文献标识码：A 文章编号：1002-6630(2009)01-0151-04 纳豆激酶是由纳豆芽孢杆菌(Bacillus subtilis Natto)产 素效应进行t 检验，选择可信度较高的因素作为重要因 生的一种具