生物学杂志 ›› 2020, Vol. 37 ›› Issue (4): 101-.doi: 10.3969/j.issn.2095-1736.2020.04.101

• 技术方法 • 上一篇    下一篇

响应面法优化假单胞菌产低温淀粉酶发酵培养基

  

  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
  • 出版日期:2020-08-18 发布日期:2020-08-10
  • 作者简介:王继莲,硕士,副教授,主要从事低温微生物学方面的研究,E-mail:wjilian0710@sina.com
  • 基金资助:
    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)

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

  1. 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
  • Online:2020-08-18 Published:2020-08-10

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。

关键词: 低温淀粉酶, 发酵培养基, 响应面法, Box-Benhnken设计

Abstract: 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.

Key words: cold- active amylase, fermentation medium, response surface methodology, Box-Benhnken design

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