丁酸梭菌MY-Z高密度发酵培养基及发酵工艺的优化

Optimization of high density fermentation medium and fermentationprocess of Clostridium butyricum MY-Z

[目的]研究丁酸梭菌的营养条件并优化发酵工艺,为与丁酸梭菌培养发酵相关的实验室研究及规模化生产工艺优化提供依据。[方法]通过单因素试验,比较8种常见廉价碳氮源对丁酸梭菌增菌的影响,对高密度发酵培养基和培养条件进行优化。[结果]丁酸梭菌MY-Z发酵最佳营养组分为:葡萄糖30.0g/L、酵母粉5.0g/L、鱼粉30.0g/L或肽粉30.0g/L、K_(2)HPO_(4)1.05g/L、MgSO_(4)·7H_(2)O1.21g/L、NaCl0.3g/L、CaCO_(3)1.5g/L、MnSO_(4)·H_(2)O0.012g/L、促生长因子0.2g/L;最佳发酵工艺为:37℃恒温培养30~36h,发酵过程中恒定pH为6.5,并在发酵10h左右进行总量为5%的连续变量补料。丁酸梭菌MY-Z在此发酵工艺下,发酵32h左右,芽孢数可达2.28×10^(9)CFU/mL。[结论]通过优化发酵工艺可有效提高发酵液菌数,从而降低生产成本,对丁酸梭菌的产业化推广与应用具有重要意义。

[Objectives]We investigated the nutritional condition for C.butyricum and optimized the fermentation process,so as to provide a basis for the laboratory research and optimization the large-scale production process related to the culture and fermentation of Clostridium butyricum.[Methods]The effects of 8 common cheap carbon and nitrogen sources on the growth of C.butyricum were compared through single factor experiment,and the high-density fermentation medium and culture conditions were optimized.[Results]The optimal nutrient components for C.butyricum MY-Z fermentation were 30.0 g/L glucose,5.0 g/L yeast powder,30.0 g/L fish meal(or 30.0 g/L peptide powder),1.05 g/L K_(2)HPO_(4),1.21 g/L MgSO_(4)·7 H_(2)O,0.3 g/L NaCl,1.5 g/L CaCO_(3),0.012 g/L MnSO_(4)·H_(2)O,and 0.2 g/L growth promoting factor.The optimal fermentation process was constant temperature culture at 37℃for 30-36 hours,constant pH 6.5 during fermentation,and continuous variable feeding with a total amount of 5%at about 10 th hour of fermentation.C.butyricum MY-Z ferments for about 32 hours under this fermentation process,and the number of spores could reach 2.28×10^(9)CFU/mL.[Conclusions]The experiment shows that the optimization of fermentation process effectively increased the number of bacteria in the fermentation liquid,thereby reducing the production cost,which is of great significance for the industrial promotion and application of C.butyricum.