Optimization of fermentation procedure for Haemophilus influenzae type bChinese Full Text
LI Gui-fan,LI Xiu-mei,WEI Wen-jin,LU Fu-ping(Key Laboratory of Industrial Fermentation of Microorganisms,Ministry of Education,College of Bioengineering,Tianjin University of Science and Technology,Tianjin 300457,China)
Abstract: 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（A550 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 controlled at 20% and 7.2 respectively,while 200 g / L glucose was supplemented 5 h after inoculation,at a rate of 3 ml / min.Compared 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 polysaccharide from 63 to 96 mg / L.The final concentrations（A550 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.
- DOI：
10.13200/j.cjb.2012.06.108.ligf.006
- Series：
- Subject：
- Classification Code：
R378.41
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