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.
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)
出处
《生物学杂志》
CAS
CSCD
北大核心
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)。
