b型流感嗜血杆菌发酵工艺的优化  

Optimization of fermentation procedure for Haemophilus influenzae type b

作　　者：李贵凡[1,2] 李秀梅[1] 魏文进[2] 路福平[1] 

机构地区：[1]天津科技大学生物工程学院教育部工业发酵微生物重点实验室,天津300457 [2]北京民海生物科技有限公司,北京102600

出　　处：《中国生物制品学杂志》2012年第6期763-766,共4页Chinese Journal of Biologicals

摘　　要：目的优化b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)的发酵工艺,以提高Hib荚膜多糖的产率。方法通过考察初始pH值和葡萄糖浓度对培养液菌体浓度(A550值)及多糖产率的影响,优化摇瓶培养条件;通过考察溶氧百分比、恒定pH值及补加葡萄糖对Hib生长和多糖产率的影响,优化20 L发酵罐的培养条件。以优化的发酵工艺连续进行3批Hib发酵,考察优化发酵工艺的稳定性。结果优化的摇瓶培养条件为:初始pH值为7.2,初始葡萄糖浓度为6 g/L。优化的发酵工艺为:溶氧百分比控制在20%,控制pH值恒定为7.2,于接种后5 h开始补加200 g/L葡萄糖,速率为3 ml/min。与优化前相比,Hib对数生长期由8 h延长至10 h左右,菌体终浓度(A550值)由1.5提高至3.4,Hib荚膜多糖产率由63 mg/L提高至96 mg/L。以优化的发酵工艺培养的3批Hib培养物菌体终浓度(A550值)在3.3～3.7之间,多糖产率在92～98 mg/L之间,稳定性较好。结论优化的发酵工艺可延长Hib对数生长期、促进菌体生长、提高荚膜多糖产率。Objective To optimize the procedure for fermentation of Haemophilus influenzae type b （Hib） and increase the yield of Hib capsular polysaccharide. Methods The effects of original pH value and glucose concentration on bacterial concentration （A 550 value） and polysaccharide yield of medium were investigated, based on which the condition for culture in shake flask was optimized. The effects of DO, constant pH value and glucose supplement on growth of Hib and yield of polysaccharide were investigated, based on which the condition for culture in 20 L fermenter was optimized. Three consecutive batches of Hib were fermented to evaluate the stability of the optimized procedure. Results The original pH value and glucose concentration for culture in shake-flask were optimized as 7. 2 and 6 g/L respectively. The fermentation procedure was optimized as follows： the DO and pH value were con- trolled at 20% and 7.2 respectively, while 200 g / L glucose was supplemented 5 h after inoculation, at a rate of 3 ml / min. Com- pared with those by the procedure before optimization, the logarithmic growth phase of Hib fermented by the optimized procedure increased from 8 h to about 10 h, while the final concentration （A550 value） of bacteria from 1. 5 to 3. 4, the yield of Hib capsular polysaecharide from 63 to 96 mg/ L. The final concentrations （A 550 value） of bacteria and yield of Hib capsular polysaccharide of three batches of Hib fermented were 3.3 - 3. 7 and 92 - 98 mg/ L respectively, indicating high stability of the optimized procedure. Conclusion By the optimized fermentation procedure, the logarithmic growth phase of Hib was prolonged, the growth of bacteria was promoted, and the yield of Hib capsular polysaccharide increased.

关 键 词：B型流感嗜血杆菌 发酵 优化 

分 类 号：R378.41[医药卫生—病原生物学] R392-33[医药卫生—基础医学]