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首页> 外文期刊>Applied Microbiology and Biotechnology >Capsular polysaccharide production by Streptococcus pneumoniae serotype 1: from strain selection to fed-batch cultivation
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Capsular polysaccharide production by Streptococcus pneumoniae serotype 1: from strain selection to fed-batch cultivation

机译：肺炎链球菌血清型1产生的荚膜多糖：从菌株选择到分批分批培养

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摘要

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.
机译：肺炎链球菌是一种人类病原体，主要通过气溶胶传播。疫苗是抵抗这种病原体的主要策略，荚膜多糖（PS）是其主要抗原。肺炎链球菌血清型1与大流行和侵袭性疾病的流行有关。这项工作的目的是筛选1型血清型菌株，以确定最佳的PS1生产者，评估生产PS1的三种蛋白,，使用实验设计（DoE）（一种用于优化的统计工具）研究培养基成分的影响，并提出一个建议。新的培养基/栽培策略。在九种菌株的烧瓶培养之后，选择了两个产生高PS1和生物量值的菌株用于生物反应器中的进一步评估，并选择ST595 / 01作为最佳PS1生产菌株。在测试的蛋白（（酪蛋白氨基酸，大豆蛋白和植物蛋白）中，植物蛋白达到了最高的PS1产量（298 mg / L）。接下来，执行DoE（2（4-1））来评估酵母提取物（YE），植物提取物，L-天冬酰胺（Asn）和L-谷氨酰胺（Gln）的作用，产生以下结果：植物提取物表现出积极的作用（p <0.05），以最大程度地生产生物量，PS1，乙酸盐和乳酸盐; YE显示出对生物量和产酸的积极影响（p <0.05）; Gln对PS1产量因子对葡萄糖的影响较小（p <0.1）; Asn仅对乙酸盐产生有影响（p <0.1）。因此，基于Phytone，YE和葡萄糖配制了新的培养基，并对分批和补料分批培养进行了评估。分批补料培养显示的生物量几乎是分批培养的2倍，PS1产量是2.5倍，PS1产量比以前报道的高8-10倍。

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