两步控制策略优化生防菌B579芽孢生成条件  

Optimization of Sporulation Condition of Biocontriol Bacteria B579 by Two-step Control Strategy

作　　者：张艳春[1] 贾钧辉[1] 郑宇[1] 陈芳[1] 曾明洁[1] 骆健美[1] 王敏[1] 

机构地区：[1]天津科技大学生物工程学院工业微生物教育部重点实验室,天津300457

出　　处：《安徽农业科学》2011年第13期7685-7687,共3页Journal of Anhui Agricultural Sciences

基　　金：天津市应用基础与前沿技术项目(09JCZDJC19100);天津科技大学科学研究基金项目(20090403)

摘　　要：[目的]优化生防菌-枯草芽孢杆菌B579芽孢生成的发酵条件,提高芽孢浓度。[方法]采用2步控制策略,即发酵的第1阶段（菌体生长阶段,0~10 h）促进细胞生长,发酵的第2阶段（菌体对数生长后期,芽孢生成阶段,10~30 h）促进芽孢生成,分别考察了培养基初始葡萄糖浓度、发酵pH、第2阶段摇床转速、第2阶段发酵温度对芽孢生成的影响。通过单因素试验确定其水平,在单因素试验基础上进行正交试验优化,确定获得高浓度芽孢的最佳发酵条件。[结果]培养基初始葡萄糖浓度是影响芽孢生成的显著因素,芽孢生成的最佳发酵条件为培养基初始葡萄糖浓度5 g/L,发酵pH7.0,第1阶段的发酵温度37℃,摇床转速180 r/min,约10 h后进入第2阶段,发酵温度提高到40℃,摇床转速调节为200 r/min。发酵30 h,最终获得的芽孢浓度为（9.43±0.15）×108CFU/ml,芽孢生成率为90.99%,分别是优化前的6.70和2.43倍。[结论]利用两步控制发酵策略可提高生防菌B579芽孢浓度,建立的发酵工艺为生防菌剂的大规模生产奠定了基础。[Objective]The aim was to optimize the germination condition for the formation of biocontrol bacteria B579 gemma and to improve the concentration of gemma.[Method] Two-step control strategies were adopted.The first germination stage facilitate the growth of cells（bacterial growth period,0~10 d）.The second stage of germination facilitates the formation of gemma（later period of the growth of germmas,10~30 h）.The initial glucose concentration,germination pH value of culture medium,the rotating speed in the second period and the influence of germination temperautre in the second period to the formation of gemma were investigated.By dint of single factor experiment to determine its level,and orthogonal test opertimization based on single factors experiment,the best germination condition for high concentration gemma was determined.[Result] The initial glucose concentration of culture medium was the significant factor that influenced the formation of gemma.The optimized germination condition for the formaiton of gemma was 5 g/L glucose concentration,7.0 germination pH value.The germination temperature in the first stage was 37 ℃,rotating speed 180 r/min.After 10 h,it stepped into second stage,when germination temperature rose to 40 ℃,the rotating speed was 200 r/min and germination lasted for 30 h.The final concentration of gemma was（9.43±0.05）×108CFU/ml.The formation rate of gemma was 90.99%,which was 6.70 and 2.43 times before optimization.[Conclusion] The two step control germination stratagies could enhance the concentration of biocontrol bacteria B579 gemma.And the formed germination craft laid basis for the large scale production of biocontrol bacteria agents.

关 键 词：枯草芽孢杆菌 生防细菌 芽孢 两步控制策略 发酵 

分 类 号：S188[农业科学—农业基础科学]