Capsular polysaccharide production by Streptococcus pneumoniae serotype 1: from strain selection to fed-batch cultivation

Autor(es)

Marthos, Bruno Vitorio ^[1] ; Silva Ferri, Anne Leticia ^[1] ; de Figueiredo, Douglas Borges ^[1] ; Zangirolami, Teresa Cristina ^[2] ; Goncalves, Viviane Maimoni ^[1]

Afiliação do(s) autor(es)

^[1] Inst Butantan, Ctr Biotecnol, Lab Bioproc, BR-05503900 Sao Paulo, SP - Brazil

^[2] Univ Fed Sao Carlos, Dept Engn Quim, BR-13565905 Sao Carlos, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Applied Microbiology and Biotechnology; v. 99, n. 24, p. 10447-10456, DEC 2015.
• Citações Web of Science: 1

Resumo

Streptococcus pneumoniae is a human pathogen largely transmitted by aerosols. Vaccines are the main strategy against this pathogen, and the capsular polysaccharide (PS) is its major antigen. S. pneumoniae serotype 1 is associated with large outbreaks and epidemics of invasive diseases. The aims of this work were to screen serotype 1 strains to identify the best PS1 producer, evaluate three peptones for PS1 production, investigate the effects of culture medium components using a design of experiments (DoE), a statistic tool for optimization, and propose a new medium/cultivation strategy. After flask cultivation of nine strains, two that produced high PS1 and biomass values were chosen for further evaluation in the bioreactor, and ST595/01 was chosen as the best PS1 producer strain. Among the peptones tested (Casamino acids, Soytone, and Phytone), the highest PS1 production (298 mg/L) was reached with Phytone. Next, DoE (2(4-1)) was performed to evaluate the effects of yeast extract (YE), Phytone, L-asparagine (Asn), and L-glutamine (Gln), yielding the following results: Phytone presented positive effects (p < 0.05) for maximum production of biomass, PS1, acetate, and lactate; YE showed positive effects for biomass and acid production (p < 0.05); Gln exerted a minor positive effect on PS1 yield factor on glucose (p < 0.1); and Asn presented only an effect on acetate production (p < 0.1). Hence, a new culture medium was formulated based on Phytone, YE, and glucose, and batch and fed-batch cultivations were evaluated. The fed-batch cultivation showed almost 2 times the biomass and 2.5 times the PS1 production as the batch culture, and 8-10 times higher PS1 production than has been previously reported. (AU)

• Processo FAPESP: 11/01262-3 - Determinação da influência do meio de armazenamento sobre a viabilidade celular e a produção de polissacarídeo capsular de Streptococcus pneumoniae
• Beneficiário: Douglas Borges de Figueiredo
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica
• Processo FAPESP: 08/05207-4 - Vacina conjugada anti-pneumocócica: estudos sobre a viabilidade de uma vacina polissacarídeo capsular - PSPA
• Beneficiário: Martha Massako Tanizaki
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 11/02805-0 - Determinação dos aminoácidos essenciais e da influência das bases nitrogenadas em cultivo contínuo de Streptococcus pneumoniae sorotipo 14
• Beneficiário: Anne Leticia Silva Ferri
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 08/10364-1 - Impacto das estratégias de cultivo de Streptococcus pneumoniae sobre a produção do polissacarídeo capsular
• Beneficiário: Viviane Maimoni Gonçalves
• Modalidade de apoio: Auxílio à Pesquisa - Regular