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生物反应器曝气条件在猪丹毒丝菌培养过程中调节生长和抗原表达。

Bioreactor aeration conditions modulate growth and antigen expression during Erysipelothrix rhusiopathiae cultivation.

作者信息

da Silva Adilson José, de Baptista-Neto Alvaro, do Carmo Cilento Maria, de Campos Giordano Roberto, Zangirolami Teresa Cristina

机构信息

Departamento de Engenharia Química, Universidade Federal de São Carlos, Rod Washington Luís, São Carlos, SP, Brazil.

出版信息

Appl Microbiol Biotechnol. 2008 May;79(1):23-31. doi: 10.1007/s00253-008-1399-0. Epub 2008 Mar 7.

Abstract

Erysipelothrix rhusiopathiae, the causative agent of swine erysipelas, was cultivated in a 5-L stirred and aerated bioreactor under different dissolved oxygen tensions (0%, 5%, and 30% of saturation) for evaluation of the influence of oxygen on cell growth as well as on the production of the main antigenic component of the vaccine against erysipelas, a 64-69 kDa protein (SpaA). The microorganism presented different growth profiles for different aeration conditions. However, at the end of the batch cultivations, similar cell concentrations were obtained under the studied conditions. In order to maximize biomass titers and antigen production, the microorganism was cultivated in fed-batch operation mode under aerobic conditions. Under this condition, there was a fivefold increase in biomass production in comparison to the results attained in batch cultivations. To follow up antigen expression, samples collected during batch cultivations were concentrated and treated with choline for antigen extraction. Antigen expression was then assessed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and by murine immunization tests. It was observed a direct influence of oxygen availability upon antigen expression, which is favored in the presence of oxygen. Analysis of the samples collected throughout the fed-batch process also revealed that antigen production is growth associated.

摘要

猪丹毒的病原体猪红斑丹毒丝菌在一个5升的搅拌式通气生物反应器中,于不同的溶解氧张力(饱和度的0%、5%和30%)下进行培养,以评估氧气对细胞生长以及对猪丹毒疫苗主要抗原成分(一种64 - 69 kDa的蛋白质(SpaA))生产的影响。该微生物在不同的通气条件下呈现出不同的生长曲线。然而,在分批培养结束时,在所研究的条件下获得了相似的细胞浓度。为了使生物量效价和抗原产量最大化,该微生物在有氧条件下以补料分批操作模式进行培养。在此条件下,与分批培养所获得的结果相比,生物量产量增加了五倍。为了跟踪抗原表达,在分批培养过程中收集的样品进行浓缩并用胆碱处理以提取抗原。然后通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳和小鼠免疫试验评估抗原表达。观察到氧气供应对抗原表达有直接影响,在有氧气的情况下抗原表达更有利。对补料分批过程中收集的样品进行分析还表明,抗原生产与生长相关。

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