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中国农学通报

中国农学通报 ›› 2016, Vol. 32 ›› Issue (17): 25-34.doi: 10.11924/j.issn.1000-6850.casb16010057

所属专题： 水产渔业

• 生物技术科学 • 上一篇    下一篇

响应面分析法在海洋生防细菌变形斑沙雷氏菌(Serratia proteamaculans)增殖培养基优化中的应用

雷 蕾,张 莉,孟祥红   

  1. 中国海洋大学 食品科学与工程学院,中国海洋大学 食品科学与工程学院,中国海洋大学 食品科学与工程学院
  • 收稿日期:2016-01-12 修回日期:2016-06-02 接受日期:2016-03-22 出版日期:2016-06-20 发布日期:2016-06-20
  • 通讯作者: 张 莉
  • 基金资助:
    青岛市科技发展计划“用于果蔬病害防治的海洋生防酵母制剂的研制和应用”(12-1-3-34-nsh)。

Optimization of Medium Composition for Acquiring the Max Growth Biomass of a Marine Bio-control Bacteria Serratia proteamaculans by Response Surface Methodology

  • Received:2016-01-12 Revised:2016-06-02 Accepted:2016-03-22 Online:2016-06-20 Published:2016-06-20

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摘要： 为快速大量地得到生防菌Serratia proteamaculans NC-5，并且提高菌株的生防效果，本文对培养基组成进行优化并对各组成成分之间的相互作用进行了深入分析。先对其培养基组成成分包括碳源、氮源、二价金属离子及NaCl、KH2PO4、酵母粉进行单因素试验，再通过Plackeet-Burman实验设计方法可知发酵培养基对NC-5的生长具有显著影响的因素为麦芽糖、胰蛋白胨和CaCl2·2H2O。根据重要影响因素的效应大小设定爬坡方向及爬坡步长进行最陡爬坡试验。最后，采用Box-Behnken试验设计3因素3水平的响应面分析试验。结果显示麦芽糖、胰蛋白胨的最佳质量浓度和CaCl2·2H2O的最佳浓度组成分别为7.68 g/L、32.88 g/L、4.672 mmol/L。经过优化后的菌株NC-5的增殖培养基的组成成分为：麦芽糖7.68 g/L、胰蛋白胨32.88 g/L、KH2PO4 0.25%、NaCl 0.50%、4.672 mmol/L CaCl2·2H2O、MgSO4·7H2O 2 mmol/L、MnSO4·1H2O 1 mmol/L、酵母粉1 g/L。采用以上最佳培养基进行培养，培养后菌株NC-5的吸光值OD600为0.821（比色时菌液稀释4倍），基本与预测的吸光值0.81652接近，并是基础培养基培养菌株后菌悬液吸光值0.675（比色时菌液稀释2倍）的2.432倍，即发酵液菌体的细胞数量提高了184.444%。以上实验说明响应面法优化得到的函数模型与实际数据较为拟合，经过优化后的培养基组成可为该生防菌的工业生产提供理论基础。

关键词: 虾夷扇贝, 虾夷扇贝, 壳性状, 体腔液, 超氧化物歧化酶, 过氧化氢酶, 过氧化物酶

Abstract: To acquire the max growth biomass and improve the biocontrol effect of Serratia proteamaculans NC-5, the culture medium composition was optimized and the interactions among the compositions were analyzed in the paper. At first, single factor experiments were conducted to acquire the best concentration of carbon, nitrogen, bivalent metallic cations, NaCl, KH2PO4 and yeast powder. Then maltose, tryptone and CaCl2·2H2O were found to significantly influence the growth of S. proteamaculans NC-5 by Plackeet-Burman Design. Steepest ascent experiment was designed by the effect proportion of the significant influencing factors. Finally, Box-Behnken Design was used to design the 3 factors and 3 levels response surface analysis. The results showed that the best concentration of maltose, tryptone and CaCl2·2H2O was 7.68 g/L, 32.88 g/L, and 4.672 mmol/L, respectively. The optimum medium was composed of 7.68 g/L maltose, 32.88 g/L tripton, 0.25% KH2PO4, 0.5% NaCl, 4.672 mmol/L CaCl2·2H2O, 2 mmol/L MgSO4·7H2O, 1 mmol/L MnSO4·1H2O and 1 g/L yeast powder after optimization of the composition of culture medium. After cultivating with the above culture medium, OD600 was 0.821 (dilution 4 times), which was closed to the prediction (0.81652), and was 2.432 times as much as that of basic medium (0.675, dilution 2 times), and cell count of fermentation broth increased by 184.444%. The results indicated that the mathematical model was fitted with the experimental data and the medium composition could be used for industrial production.