一株枯草芽孢杆菌的鉴定及液体发酵工艺优化  

作　　者：胡红伟 段明房 闫凌鹏 邢俊德[2] 麻啸涛 李自茹 陈茹茹 杨京娥 

机构地区：[1]山西大禹生物工程股份有限公司,山西芮城044600 [2]太原理工大学化学工程学院,山西太原030024

出　　处：《中国饲料》2017年第5期13-19,共7页China Feed

基　　金：山西省重点研发计划(201603D221026-3)

摘　　要：从鸡肠道筛选出一株菌,命名为DY032,经16S r DNA分子鉴定和系统发育分析,该菌种为枯草芽孢杆菌。在摇瓶发酵条件下,采用单因素试验,对菌株DY032的培养基配方进行优化,确定了影响发酵液菌体数和芽孢数的5个因素,分别为淀粉、鱼粉、豆饼粉、MnSO_4·H_2O和CaCO_3,其适宜浓度分别为20、10、10、0.05、0.05 g/L,进一步通过L_(16)(4~5)正交试验确定以上5因素的最佳配比。为放大培养,在最优培养基基础上,利用10 L发酵罐,采用L_9(3~4)正交试验设计,以芽孢数为指标,对菌株DY032的培养温度、pH值、搅拌转速、通气量4个条件进行优化。结果表明:菌株DY032培养基最优配方为:淀粉16 g/L,鱼粉10 g/L,豆饼粉10 g/L,Mn SO4·H2O 0.05 g/L,Ca CO30.03 g/L。最优发酵罐培养条件为:培养温度37℃,pH值6.5,转速200 r/min,通气量5 L/min,最终芽孢数可达51.95×108cfu/m L。A bacterium named DY032 screened from gut of chicken, designated strain DY032 , throngh the 16S rDNA sequence molecular identificaton and phylogenetic analysis,which was indicated Bacillus subtillis .Single factor test was used to optimize the fermentated medium of strain DY032.Based on the results of single factor test, five important factors that influence the viable count and spore were determined.Appropriate concentration of starch, fishmeal, soybean cake powder, MnSO4 ？ H O and CaC〇 3 were 20,10,10,0.05,0.05 g/L, respectively.To determine the best proportion of the above 5 factors,Lm （45 ）orthogonal test were used.Based on the the optimum medium, the culture temperature, pH, stirring speed and air flow rate during fermentation were optimized by the L9 （34 ）orthogonal test in 10 L bioreactor, spores were considered as index.The results showed that the optimum medium was as follows：starch 16 g/L , fishmeal 10 g/L , soybean cake powder 10 g/ L, MnSO4 , H2O0.05 g/ L, CaCO3 0.03 g/ L.The optimal culture conditions were culture temperature of 37℃, pH of 6.5,stirring speed of 200 r/min, and air flow rate of 5 L/min.Under these optimization conditions, the number of spore was up to 51.95×10^8 cfu/mL.

关 键 词：菌株DY032 16SrDNA 优化 

分 类 号：S816.3[农业科学—饲料科学]