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CpG基序重组菌发酵培养基的优化

Optimization of the Fermentation Medium of the Strain with the CpG Motifs Recombinant Plasmid

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【作者】 李华周叶明强黄志丰邝哲师赵祥杰姜增固马苗鹏王候光徐臣超曹丁张玲华

【Author】 LI Hua-zhou1,YE Ming-qiang3,HUANG Zhi-feng2,KUANG Zhe-shi3,ZHAO Xiang-jie3,JIANG Zeng-gu1, MA Miao-peng2,WANG Hou-guang1,XU Chen-chao2,CAO Ding2,ZHANG Ling-hua2*(1.Guangzhou Fine Breed Swine Farm,Guangzhou 510540,China; 2.College of Life Science,South China Agricultural University,Guangzhou 510642,China; 3.Bio-Tech Research Institute,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,China)

【机构】 广州市良种猪场广东省农科院农业生物技术研究所华南农业大学生命科学学院

【摘要】 目的:为提高CpG基序重组菌的发酵水平,降低CpG重组质粒的生产成本,运用响应面法优化CpG重组菌发酵培养基。方法:利用Plackett-Burman试验筛选出影响发酵水平的3个最显著因素:糖蜜、玉米浆和工业蛋白胨。然后用响应面设计试验并优化得到最显著因素的最佳浓度,并进一步通过发酵试验验证优化后的培养基。结果:得到一组具有较高菌浓及质粒产量且价格低廉的发酵用培养基:糖蜜4.5g/L,玉米浆8.5g/L,工业蛋白胨8.5g/L,甘油10mL/L,KH2PO41.5g/L,K2HPO42.3g/L,MgSO4.7H2O 0.25g/L,在此培养条件下,OD600实测值为0.6771,理论值为0.6643,两者接近,与标准LB培养基相比,质粒产量提高了15%左右。结论:最终筛选到的培养基具有较高的性价比。

【Abstract】 Objective:In order to improve the production of the CpG motifs recombinant stain and lower the cost,the response surface method was applied to optimize the fermentation medium.Method:Firstly,Plackett-Burman design was undertaken to determine the three significant factors affecting strain production as follows:molasses,corn syrup and peptone.the response surface design was used to optimize the three critical factors and the optimum levels,furthermore the fermentation was testified the optimum medium.Result:The optimum fermentation medium was 4.5g/L molasses,8.5g/L corn syrup,8.5g/L peptone,10mL/L glycerol,1.5g/L KH2PO4,2.3g/L K2HPO4,0.25g/L MgSO4·7H2O.Under such conditions,the predicted OD600 was 0.664 3,which was in excellent agreement with the experimental result 0.6771,about 15% higher than the plasmid production under the initial LB medium.Conclusion:The final optimum medium was identified to make lower costs and higher yield of strain and plasmid yield.

【基金】 广州市科技支撑计划项目(“抗仔猪断奶应激腹泻CpG制剂关键技术研究及示范”,编号:2010Z1-E311);广东省产学研项目(“分泌型毕赤酵母发酵制备饲用抗菌肽PR39的研究及应用”,编号:2011B090300020)资助
  • 【分类号】TQ920.6
  • 【下载频次】106
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