响应面法优化假单胞菌产低温淀粉酶发酵培养基  

Medium optimization of cold-active amylase by Pseudomonas sp.using response surface methodology

作　　者：王继莲 任羽 李明源 WANG Ji-lian;REN Yu;LI Ming-yuan(School of Biological and Geographic Sciences,Kashi University,Key Laboratory of Ecology and Biological Resources in Yarkand Oasis of Education,Xinjiang Uygur Autonomous Region,Kashgar 844006,China)

机构地区：[1]喀什大学生命与地理科学学院叶尔羌绿洲生态与生物资源研究高校重点实验室,喀什844006

出　　处：《生物学杂志》2020年第4期101-105,共5页Journal of Biology

基　　金：新疆维吾尔自治区高校科研计划项目(XJEDU2016S072);新疆维吾尔自治区自然科学基金(2016D01B011)。

摘　　要：对前期分离的产低温淀粉酶菌株假单胞菌Pseudomonas sp.D5-2进行发酵培养基优化,以进一步提升其产酶能力。通过单因素实验筛选最佳碳源、氮源和无机盐种类,在此基础上利用响应面分析法,采用Box-Benhnken设计建立数学模型,分析显著性因素的最佳水平及交互作用,得到最佳发酵组分配伍。结果表明,Pseudomonas sp.D5-2发酵产酶的最适碳源、氮源及无机盐分别为乳糖、胰蛋白胨和硫酸铜。二次回归模型响应面分析显示,当乳糖浓度为19.715 g/L,胰蛋白胨浓度14.191 g/L,硫酸铜浓度2.417g/L时,预测最高酶活力为98.876 U/mL。在上述条件下,实际酶活力为99.2 U/mL,与模型理论值吻合度较高。利用Box-Benhnken响应面设计法优化Pseudomonas sp.D5-2产低温淀粉酶发酵培养基组分方便可行,为后续深入阐释低温酶催化机理和工业应用提供指导。The fermentation medium for the clutivation of cold-active amylase producing strain Pseudomonas sp.D5-2,which was isolated in previous studies,was optimized to improve its enzyme production capacity.The effect of medium factor was determined by single factor test.Then,base on surface methodology,a Box-Benhnken design was used to identify the optimal level and interaction of significant factors to obtain the best combination.The results revealed that lactose,peptone and Cu2+were important factors affecting fermentation.The optimal conditions for producing of the maximal amylase activity by strain pseudomones sp.were as follows:19.715 g/L lactose,14.191 g/L peptone and 2.417 g/L CuSO4.Under the conditions,the highest amylase activity was predicted to be 98.876 U/mL,and the actual value was 99.2 U/mL,which was highly consistent with the theoretical model.The Box-Benhnken response surface method is feasible to optimize the fermentation medium of D5-2,which can provide guidance for the explanation of catalytic mechanism and industrial application of cold-active enzyme.

关 键 词：低温淀粉酶 发酵培养基 响应面法 Box-Benhnken设计 

分 类 号：Q939.97[生物学—微生物学]