Optimization of medium formulation and seed conditions for expression of mature PsaA (pneumococcal surface adhesin A) in Escherichia coli using a sequential experimental design strategy and response surface methodology

dc.creatorLarentis, AL
dc.creatorMonteiro, JF
dc.creatorNicolau, Q
dc.creatorArgondizzo, APC
dc.creatorGaller, R
dc.creatorRodrigues, MI
dc.creatorMedeiros, MA
dc.date2012
dc.dateJUN
dc.date2014-08-01T18:37:47Z
dc.date2015-11-26T18:06:01Z
dc.date2014-08-01T18:37:47Z
dc.date2015-11-26T18:06:01Z
dc.date.accessioned2018-03-29T00:48:14Z
dc.date.available2018-03-29T00:48:14Z
dc.identifierJournal Of Industrial Microbiology & Biotechnology. Springer Heidelberg, v. 39, n. 6, n. 897, n. 908, 2012.
dc.identifier1367-5435
dc.identifierWOS:000304402200010
dc.identifier10.1007/s10295-012-1099-8
dc.identifierhttp://www.repositorio.unicamp.br/jspui/handle/REPOSIP/81673
dc.identifierhttp://repositorio.unicamp.br/jspui/handle/REPOSIP/81673
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/1293260
dc.descriptionPsaA, a candidate antigen for a vaccine against pneumonia, is well-conserved in all Streptococcus pneumoniae serotypes. A sequence of two-level experimental designs was used to evaluate medium composition and seed conditions to optimize the expression of soluble mature PsaA in E. coli. A face-centered central composite design was first used to evaluate the effects of yeast extract (5 and 23.6 g/L), tryptone (0 and 10 g/L), and glucose (1 and 10 g/L), with replicate experiments at the central point (14.3 g/L yeast extract, 5 g/L tryptone, 5.5 g/L glucose). Next, a central composite design was used to analyze the influence of NaCl concentration (0, 5, and 10 g/L) compared with potassium salts (9.4 g/L K2HPO4/2.2 g/L KH2PO4), and seed growth (7 and 16 h). Tryptone had no significant effect and was removed from the medium. Yeast extract and glucose were optimized at their intermediate concentrations, resulting in an animal-derived material-free culture medium containing 15 g/L yeast extract, 8 g/L glucose, 50 mu g/mL kanamycin, and 0.4% glycerol, yielding 1 g/L rPsaA after 16 h induction at 25A degrees C in shake flasks at 200 rpm. All the seed age and salt conditions produced similar yields, indicating that no variation had a statistically significant effect on expression. Instead of growing the seed culture for 16 h (until saturation), the process can be conducted with 7 h seed growth until the exponential phase. These results enhanced the process productivity and reduced costs, with 5 g/L NaCl being used rather than potassium salts.
dc.description39
dc.description6
dc.description897
dc.description908
dc.descriptionFundacao Oswaldo Cruz (Fiocruz)
dc.languageen
dc.publisherSpringer Heidelberg
dc.publisherHeidelberg
dc.publisherAlemanha
dc.relationJournal Of Industrial Microbiology & Biotechnology
dc.relationJ. Ind. Microbiol. Biotechnol.
dc.rightsfechado
dc.rightshttp://www.springer.com/open+access/authors+rights?SGWID=0-176704-12-683201-0
dc.sourceWeb of Science
dc.subjectDesign of experiments (DoE)
dc.subjectModeling
dc.subjectOptimization
dc.subjectResponse surface methodology (RSM)
dc.subjectStatistical experimental design
dc.subjectStatistical Experimental-design
dc.subjectActivating Protein-production
dc.subjectCell-density Culture
dc.subjectStreptococcus-pneumoniae
dc.subjectIntranasal Immunization
dc.subjectProtective Immunity
dc.subjectPhytase Production
dc.subjectFusion Protein
dc.subjectVaccines
dc.subjectMice
dc.titleOptimization of medium formulation and seed conditions for expression of mature PsaA (pneumococcal surface adhesin A) in Escherichia coli using a sequential experimental design strategy and response surface methodology
dc.typeArtículos de revistas


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