羊源丁酸梭菌HDRyYB1发酵工艺的优化

Fermentation optimization of Clostridium butyricum HDRyYB1 isolated from sheep

摘要:

【目的】目前，国内外鲜有关于羊源丁酸梭菌的报道。本课题选用羊源丁酸梭菌HDRyYB1为研究对象，对其发酵工艺进行优化，为该菌株作为饲料添加剂应用于畜牧业生产奠定基础。【方法】采用Plackett-Burman (PB)试验设计法和响应面法分析并优化显著影响HDRyYB1菌株发酵液中芽胞数的培养基成分。【结果】发酵培养基中的面粉浓度、鱼粉浓度和米粉浓度显著影响发酵液中的芽胞数，优化后的发酵培养基组分(质量体积比)为：面粉3.72%、鱼粉0.90%、米粉3.96%、酵母粉0.60%、NaCl 0.19%、MgSO4·7H2O 0.19%、KH2PO4 0.01%、NaHCO3 0.01%、CaCO3 0.48%；培养参数为：37 °C，初始pH为7.2?7.4，瓶装量100/250，接种量3%。在此条件下，HDRyYB1菌株发酵完全(18 h)的芽胞数为1.478×108 CFU/mL，是优化前的2.7倍。【结论】HDRyYB1菌株发酵培养基得到了优化，优化后的培养基可用于后期的扩大发酵试验，验证其在实践生产中的应用价值。

关键词:丁酸梭菌，发酵工艺，响应面分析法，羊源

Abstract:

[Objective] There is almost no report about the sheep-derived Clostridium butyricum in the world at present. It is the study that optimized of liquid fermentation for sheep-derived Clostridium butyricum HDRyYB1. This study sets the foundation for Clostridium butyricum HDRyYB1 industrial product as feed additive in livestock. [Methods] We used the Plackett-Burman (PB) design and response surface methodology (RSM) though determing the spore number of Clostridium butyricum HDRyYB1 in this report. [Results] The results showed that the concentration of wheat flour, rice flour, fish meal had remarkable effect on the spore number. The optimum medium for incubating the Clostridium butyricum HDRyYB1 was composed of 3.72% wheat flour (w/v), 0.90% fish meal (w/v), 3.96% rice flour (w/v), 0.60% yeast powder (w/v), 0.19% NaCl (w/v), 0.19% MgSO4·7H2O (w/v), 0.01% KH2PO4 (w/v), 0.01% NaHCO3 (w/v), 0.48% CaCO3 (w/v), at 37 °C, pH 7.2?7.4, bottled in 100/250, inoculated quantity 3%. Under this condition, the spore number was 1.478×108 colony-forming units (CFU)/mL after fermented completely in 18 h, which was 2.7 times than before. [Conclusion] These results indicate that the fermentation medium has been optimized, it can be used in the expanded fermentation experiment to verify its application value in production practice.

Key words:Clostridium butyricum, Fermentation, RSM, Sheep-derived