Monday, July 27, 2009
P24
Optimization for Capsular Polysaccharide Production by Streptococcus pneumoniae Type 3 and Conjugation of Flagellin
Sheng-De Jin, School of Biological Sciences and Technology, Chonnam National University, 300 Yongbong-dong, Gwangju, 500-757, South Korea, Seung-Jin Jung, Interdisciplinary Program of Graduate School for Bioenergy & Biomaterials, 300 Yongbong-dong, Gwangju, South Korea, Young-Min Kim, School of Biological Sciences and Technology, The second stage of BK21, Korea Ministry of Education, The Research Institute for, Chonnam National University, 300 Yongbong-dong, Gwangju, 500-757, South Korea, Hee-Kyoung Kang, The Research Institute for Catalysis, Chonnam National University, 300 Yongbong-dong, Gwangju, 500-757, South Korea, and Doman Kim, School of Biological Sciences and Technology, Interdisciplinary Program of Graduate School for Bioenergy & Biomaterials, Chonnam National University, 300 Yongbong-dong, Gwangju, 500-757, South Korea.
The capsular polysaccharides (CPS) of Streptococcus pneumoniae are essential components in virulence that are necessary to resist host phagocytic mechanisms. To overcome this problem, many researchers were focused on developing specific pneumococcal polysaccharide as vaccine. Among the 90 serotypes, the newly interested Streptococcus pnemoniae type 3 was studied in this study. Response surface methodology (RSM) was used to optimize the fermentation conditions for enhancing the capsular polysaccharide production by the effects of five-level-three-factors and their mutual interactions. A total of 20 experiments conducted in an 8 l bioreactor were designed to assess fermentation pH (X1), supplemented glucose concentration (X2), and stirring rate (X3). The predicted highest capsular polysaccharide production by the obtained optimization model equation was 256.14 mg/l at the following optimal conditions: pH was controlled at 7.5, the stirring rate was maintained at 180 rpm, and the concentration of supplemented glucose concentration was 1% (w/v). The validity of the response model was confirmed by the observation of good agreement between the predicted and experimental results. Finally, the maximum amount of CPS obtained was 255.03 ± 2.23 mg/l. To improve the immunogenicity of Type 3 CPS, flagellin b (FlaB) was conjugated to CPS.
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