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Master's Dissertation
DOI
10.11606/D.87.2011.tde-13092011-152106
Document
Author
Full name
Verônica Maria Rodege Gogola
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2011
Supervisor
Committee
Gonçalves, Viviane Maimoni (President)
Lucarini, Adriana Celia
Tonso, Aldo
Title in Portuguese
Cultivo contínuo de Streptococcus pneumoniae sorotipo 14: metabolismo e cinética de produção de polissacarídeo capsular.
Keywords in Portuguese
Streptococcus (metabolismo)
Biomassa
Glicose
Polissacarídeos bacterianos
Reatores bioquímicos
Vacinas
Abstract in Portuguese
O polissacarídeo capsular (PS) é o principal fator de virulência do S. pneumoniae e é a base para sua classificação em sorotipos. O objetivo deste trabalho foi estudar o metabolismo de S. pneumoniae sorotipo 14 em cultivo contínuo e, para tanto, foi desenvolvido um ELISA de captura para quantificação do PS produzido (PS14) e a melhor cepa produtora foi escolhida. Em cultivos descontínuos em biorreator foram observados rendimentos semelhantes para crescimento e PS14 entre meio complexo e meio quimicamente definido. No entanto, diferenças no requerimento de vitaminas foram verificadas entre cultivos em frasco e biorreator, relacionadas à disponibilidade de O2. Em cultivos contínuos observou-se que: a produção de PS14 foi associada ao crescimento celular, a glicose foi o substrato com maior impacto sobre a produção de PS14 e os fatores de conversão sobre glicose e biomassa foram maiores sob limitação de glicose/colina/glutamina; o que permite concluir que estratégias de produção de PS14 devem focar na alta densidade celular e na condição de limitação de nutrientes.
Title in English
Continuous cultivation of Streptococcus pneumoniae serotype 14: metabolism and kinetics of capsular polysaccharide production.
Keywords in English
Streptococcus (metabolism)
Bacterial polysaccharides
Biochemical reactors
Biomass
Glucose
Vaccines
Abstract in English
The capsular polysaccharide (PS) is the most important virulence factor of S. pneumoniae and is also the basis for its classification in serotypes. This study aimed to evaluate the metabolism of S. pneumoniae serotype 14 in continuous cultivation. In order to do this, a Sandwich ELISA was developed to quantify the PS (PS14) produced and the best producer strain was selected. Similar growth and PS14 yields were observed in batch cultures conducted with complex and chemically defined media. Differences in vitamin requirements were verified in flask and batch cultivation, associated to O2 availability. In continuous cultivation, the PS14 production was associated to cell growth, the glucose was the substrate with more impact on PS14 production and in limitation of glucose/choline/glutamine the yield factors per glucose consumed and per biomass produced increased. These results showed that to improve PS14 production is necessary to adopt strategies that focuses in high cell density in addition to conditions of substrate limitation.
 
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Publishing Date
2011-09-22
 
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