响应面法优化重组毕赤酵母菌表达β-甘露聚糖酶的诱导条件  

Optimization of Expression Conditions of β-mannanase Induced by Recombinant Pichia pastoris by Response Surface Methodology

作　　者：李慧玲[1,2] 刘祖艳[1] 赵敏[1] 

机构地区：[1]东北林业大学生命科学学院,黑龙江哈尔滨150040 [2]黑龙江中医药大学药学院,黑龙江哈尔滨150040

出　　处：《安徽农业科学》2014年第11期3153-3155,3198,共4页Journal of Anhui Agricultural Sciences

摘　　要：[目的]对重组毕赤酵母菌表达β-甘露聚糖酶的诱导条件进行优化。[方法]以重组毕赤酵母GS115为研究对象,通过单因素试验确定最佳产酶条件,并采用响应面试验确定3个因素的最佳水平。[结果]最佳产酶条件为:培养温度28℃,培养pH 6.5,摇床转速210 r/min,培养基装液量100 ml。响应面试验确定3个因素即装液量、甲醇添加量和pH最佳值分别为103.18 ml、1.77%和6.69;在此条件下,β-甘露聚糖酶的活力最大值为4 917.5 U/ml。[结论]验证试验证明模型预测值准确、可靠,为重组毕赤酵母菌的合理开发利用提供了依据。[Objective] To optimize the induction conditions of the recombinant Pichia pastoris β-mannanase.[Method] With recombinant Pichia pastoris GS115 as study object,the optimal enzyme producing conditions and three factors optimal level were determined by single factor experiment and response surface experiment respectively.[Result] The optimum enzyme producing conditions are：temperature 28 ℃,pH 6.5,rotating speed 210 r/rmin,volume of medium 100 ml.Response surface methodology was used to optimize the above three factors（volume of medium,methanol concentration,pH of medium）with the optimum value 103.18 ml,1.77％,6.69,respectively.The β-mannanase activity would reach the highest value（4 917.5 U/ml） under the optimized fermentation conditions.[Conclusion] The predicted values from the response surface methodology were proved to be correct and reliable by verification test,which will provide a reference for appropriate utilization and development of recombinant Pichia pastoris.

关 键 词：重组毕赤酵母 Β-甘露聚糖酶 响应面 

分 类 号：S182[农业科学—农业基础科学]