摘要
对枯草芽孢杆菌(Bacillus subtilis)CICC20034利用合成培养基液体发酵产脂肪酶的条件进行了优化。首先采用单因子实验筛选出最适诱导剂为三丁酸甘油酯,氮源为尿素,碳源为葡萄糖,无机盐为MgSO4。在此基础上,利用Plackett-Burman设计对影响产酶因素的效应进行评价,筛选出具有显著效应的三丁酸甘油酯、尿素、KH2PO4和培养基起始pH值4个最显著的因素。用最陡爬坡路径逼近最大产酶区域后,利用响应面中心组合设计对显著因素进行优化,获得最适合成培养基组分为:葡萄糖8g/L,尿素8.57g/L,三丁酸甘油酯2.62%,KH2PO42.59g/L,MgSO4.7H2O0.5g/L,TritonX-1000.5g/L,pH9.47。优化后的B.subtilis CICC 20034胞外脂肪酶活力达0.483U/ml,比初始酶活力0.072U/ml提高了6.7倍。
The fermentation conditions of lipase production by Bacillus subtilis CICC 20034 were optimized. Initially, the most suitable intruder tributyrin, nitrogen source urea, carbon glucose, and MgSO4 were selected aceording to single factorial experiments respectively. Based on the result, screening methodology Plackett-Burman design was used to evaluate the effects of twelve factors related to lipase production and four statistically significant factors tributyrin, urea, KH2PO4 and pH were selected. The path of steepest ascent was used to approach the optimal region of lipase production subsequently. Then, the optimal combined concentration for maximum enzyme activity were further optimized by response surface methodology and determined as follows： tributyrin 2. 62% , urea 8.57g/L, KH2PO42.59g/L and pH 9.47. The optimization of culture conditions of B. subtilis CICC 20034 led to a 6.7-fold increase in lipase production relative to initial result 0. 072 /ml, which indicate that single factor in combination with response surface methodology, is an effeetive method for optimization of lipase production conditions by B. subtilis CICC 20034.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2010年第8期100-105,共6页
China Biotechnology
基金
国家自然科学基金重点项目(20936002)
国家自然科学基金(20906049)资助项目
