Improved capsular polysaccharide production by Streptococcus pneumoniae serotype 14 using continuous cultivation

Gogola-Kolling, Verônica Maria Rodege; Zanardo, Rafaela Tais; Carmo, Talita Souza; Zampoli, Natália Dalfré; Figueiredo, Douglas Borges De; Gonçalves, Viviane Maimoni

doi:10.1016/j.bej.2014.07.007

Cited by 6 publications

(16 citation statements)

References 34 publications

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“…Means and standard deviations were obtained from three independent fed-batch experiments Table 6 Maximum specific growth rate and total amount of biomass, lactate, acetate, and PS1 produced in batch and fed-batch cultivations of S. pneumoniae serotype 1 using the new culture medium Calculations are based on final volume of 5 L carbon sources other than glucose. Acetate production could also be related to the higher biomass concentration obtained with this strain compared to other pneumococci (Liberman et al 2011;Leal et al 2011;Gogola-Kolling et al 2014), as acetate formation is energetically favorable, yielding three ATP molecules instead of two when lactic acid is the final product (Carvalho et al 2013). …”

Section: Discussionmentioning

confidence: 95%

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

Marthos

Ferri

Figueiredo

et al. 2015

Appl Microbiol Biotechnol

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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.

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Section: Discussionmentioning

confidence: 95%

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

Marthos

Ferri

Figueiredo

et al. 2015

Appl Microbiol Biotechnol

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“…; Gogola‐Kolling et al . ); however, cell wall polysaccharides are more difficult to separate from the biomass. Due to the constant diffuse of exocellular polysaccharides into the culture medium their separation is much easier and determines their bigger impact for biotechnological industry.…”

Section: Discussionmentioning

confidence: 99%

“…; Gogola‐Kolling et al . ). A main disadvantage of EPS production by continuous cultivation at mesophilic temperatures is the high viscosity of culture liquid due to accumulation of the synthesized EPS.…”

Section: Introductionmentioning

confidence: 97%

Continuous cultivation of a thermophilic bacteriumAeribacillus pallidus418 for production of an exopolysaccharide applicable in cosmetic creams

et al. 2015

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EPSs from thermophilic micro-organisms are of special interest due to the advantages of the thermophilic processes and nonpathogenic nature of the polymer molecules. However, their industrial application is hindered by the comparatively low biomass and correspondingly EPS yield. Suggested continuous approach for EPS could have an enormous economic potential for an industrial scale production of thermophilic EPSs.

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“…However, the relative purity was not reached after 60% ethanol precipitation and an additional chromatographic step was necessary (Figure 1). This could be a result of the greater contamination of the starting material with intercellular impurities due to pneumococcal autolysis that occurs in batch, but is avoided in continuous cultivation (Gogola-Kolling et al, 2014). Furthermore, complex medium also leads to a starting material with higher concentration of contaminants than defined medium.…”

Section: Purification Strategymentioning

confidence: 99%

DEVELOPMENT OF A NEW PROCESS FOR PURIFICATION OF CAPSULAR POLYSACCHARIDE FROM Streptococcus pneumoniae SEROTYPE 14

Zanardo

Ferri

Figueiredo

et al. 2016

Braz. J. Chem. Eng.

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-The main virulence factor of Streptococcus pneumoniae is the capsular polysaccharide (PS), which is the antigen of all current vaccines that are prepared with PS purified from serotypes prevalent in the population. In this work, three purification strategies were evaluated and a new process was developed for purification of serotype 14 PS (PS14), responsible for 39.8% of diseases in children of 0-6 years old in Brazil. The developed method consists of cell separation by tangential microfiltration, concentration of the microfiltrate by tangential ultrafiltration (50 kDa), diafiltration in the presence of sodium dodecyl sulfate using a 30 kDa ultrafiltration membrane, precipitation with 5% trichloroacetic acid, precipitation with 20% and 60% ethanol, and anion exchange chromatography. The required purity regarding nucleic acids (≤ 2%) and proteins (≤ 3%) was achieved, resulting in a relative purity of 439 mg PS14/mg nucleic acids and 146 mg PS14/mg proteins. The final polysaccharide recovery was 65%, which is higher than the recovery of the majority of processes described in the literature.

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Improved capsular polysaccharide production by Streptococcus pneumoniae serotype 14 using continuous cultivation

Cited by 6 publications

References 34 publications

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

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

Continuous cultivation of a thermophilic bacteriumAeribacillus pallidus418 for production of an exopolysaccharide applicable in cosmetic creams

DEVELOPMENT OF A NEW PROCESS FOR PURIFICATION OF CAPSULAR POLYSACCHARIDE FROM Streptococcus pneumoniae SEROTYPE 14

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