响应面法优化重组枯草芽孢杆菌产脂肪氧合酶条件  

OPTIMIZATION OF CONDITIONS FOR LIPOXYGENASE PRODUCTION FROM RECOMBINANT Bacillus subtilis BY RESPONSE SURFACE METHODOLOGY

作　　者：章栋梁[1] 钟蔚[1] 张充[1] 吕凤霞[1] 别小妹[1] 陆兆新[1] 

机构地区：[1]南京农业大学食品科技学院,江苏南京210095

出　　处：《核农学报》2012年第2期324-329,共6页Journal of Nuclear Agricultural Sciences

基　　金：国家自然科学基金项目(31071605)

摘　　要：采用Plackett-Burman设计法,对影响重组枯草芽孢杆菌发酵产脂肪氧合酶的7个相关因子进行筛选,并建立枯草芽孢杆菌产脂肪氧合酶的最佳工艺。结果表明对重组菌发酵产酶具有显著影响的因子是乳糖、发酵时间和发酵温度。依据Plackett-Burman设计法的筛选结果,进一步采用响应面中心组合设计(Central Composite Design)对显著影响因子的最佳水平和交互作用进行研究,通过模型方程的3-D图和等高线图发现,当乳糖浓度为16.9g/L、发酵时间为88.49 h、发酵温度为29.71℃和装液量为61.31ml时,出现酶活最大预测值164.90 U/ml,验证试验证实了在上述条件下得出的酶活为167.32U/ml,与预测值高度吻合。优化后的培养基和培养条件使得重组脂肪氧合酶产量较优化前提高了2.37倍。Influences of seven related factors on productivity of lipoxygenase by recombinant Bacillus subtilis were studied by the method of Plackett-Burman design. As a result, the lactose, fermentation time and temperature were found to be the most important factors. Meanwhile, a three-level four-factor central composite design was applied to optimize the levels of the chosen variables and their interaction. By analysis of the 3-D plots and corresponding plots, while the lactose concentration was 16.9g/L, fermentation time 88.49h, temperature 29.71~C and culture volume was 61.31ml, enzyme activity reached the maximum value 164.90U/ml. Verification experiments under the above conditions gave the enzyme activity of 167.32U/ml, which showed high homology with predictive value. The yield of lipoxygenase was 2.37 times higher than before in the optimized fermentation conditions.

关 键 词：脂肪氧合酶 枯草芽孢杆菌 条件优化 Plackett.Burman设计 响应曲面法 

分 类 号：TQ920.1[轻工技术与工程—发酵工程]