摘要
为获得产CpxP蛋白枯草芽孢杆菌ZY1液体发酵的最佳条件并制备微生态菌剂,采用单因素试验与响应面实验对ZY1进行发酵工艺优化研究。结果表明:摇瓶发酵培养基最佳碳源、氮源、无机盐分别为淀粉、豆粕、NaCl;最优培养基配方为淀粉2.12 g/L、豆粕5.90 g/L、NaCl 5.21 g/L,活菌数达到9.63×10^(8)CFU/mL,比培养基优化前提高了11.04倍;摇瓶发酵条件最佳初始pH值、温度、转速、接种量分别为7.0、37℃、200 r/min、2%,活菌数达1.05×10^(9)CFU/mL,比培养基优化后提高了9.0%;按10%接种量基于5 t发酵罐体系进行菌剂中试生产过程中,ZY1活菌数达到1.05×10^(11)CFU/mL,比摇瓶优化后提高了100倍;发酵结束时芽孢率达到90%,经喷雾干燥制备出菌剂10^(12)kg,成品检测活菌数为8.5×10^(9)CFU/g。研究结果可为进一步开展CpxP蛋白在动物细菌性腹泻病生物防治功能方面的研究提供基础材料。
In order to obtain the optimal conditions for liquid fermentation of Bacillus subtilis ZY1 producing CpxP protein and prepare microecological bactericides,the fermentation process of Bacillus subtilis ZY1 was optimized by single factor test and response surface test.The results show that the optimal carbon source,nitrogen source and inorganic salt of fermentation medium are starch,soybean meal and NaCl,respectively.The optimal medium formulation is starch at 2.12 g/L,soybean meal at 5.90 g/L,and NaCl at 5.21 g/L,with a viable bacteria count of 9.63×10^(8)CFU/mL,which is 11.04 times higher than that before the medium optimization.The optimum initial pH value,temperature,rotation speed and inoculation amount for shake flask fermentation are 7.0,37℃,200 r/min and 2%,respectively.The viable bacteria number is 1.05×10^(9)CFU/mL,which is 9.0%higher than that of optimized medium.During the pilot production process of the microecological bactericides based on 5 t fermentation tank system with 10%inoculated quantity,the count of viable ZY1bacteria reaches 1.05×10^(11)CFU/mL,which is 100 times higher than that after shaker optimization.When the spore rate reaches 90%at the end of fermentation,10^(12)kg bactericides are prepared by spray drying,and the count of viable bactericide is 8.5×10^(9)CFU/g.The research results can provide basic materials for further research on the biocontrol function of CpxP protein in animal bacterial diarrhea disease.
作者
董荣荣
张焕
于欣然
刘浩
周晓辉
DONG Rongrong;ZHANG Huan;YU Xinran;LIU Hao;ZHOU Xiaohui(School of Food and Biology,Hebei University of Science and Technology,Shijiazhuang,Hebei 050018,China;Department of Asset and Laboratory Management,Hebei University of Science and Technology,Shijiazhuang,Hebei 050018,China)
出处
《河北科技大学学报》
北大核心
2025年第1期100-107,共8页
Journal of Hebei University of Science and Technology
基金
国家药典委员会科研项目(2021Y04)
河北省自然科学基金生物医药联合基金重点项目(C2020208020)
河北省农业科技成果转化资金项目(202460101030040)。
关键词
蛋白质工程
CpxP蛋白
枯草芽孢杆菌
发酵优化
响应面
中试生产
protein engineering
CpxP protein
Bacillus subtilis
optimization of fermentation
response surface
pilot production
