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dc.contributor.CRUESPUniversidade Estadual de Campinaspt_BR
dc.typeArtigo de periódicopt_BR
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 methodologypt_BR
dc.contributor.authorLarentis, ALpt_BR
dc.contributor.authorMonteiro, JFpt_BR
dc.contributor.authorNicolau, Qpt_BR
dc.contributor.authorArgondizzo, APCpt_BR
dc.contributor.authorGaller, Rpt_BR
dc.contributor.authorRodrigues, MIpt_BR
dc.contributor.authorMedeiros, MApt_BR
unicamp.authorLarentis, Ariane Leites Monteiro, Julia Fabiana Correa Argondizzo, Ana Paula Galler, Ricardo Medeiros, Marco Alberto Fundacao Oswaldo Cruz Fiocruz, VDTEC Vice Diretoria Desenvolvimento Tecnol, Lab Tecnol Recombinante LATER, BR-21040360 Rio De Janeiro, RJ, Brazilpt_BR
unicamp.authorRodrigues, Maria Isabel Cidade Univ Zeferino Vaz, Univ Estadual Campinas UNICAMP, Fac Engn Alimentos, BR-13083970 Campinas, SP, Brazilpt_BR
unicamp.authorRodrigues, Maria Isabel PROTIMIZA, Consulting & Training Expt Design & Proc Optimiza, Campinas, SP, Brazilpt_BR
dc.subjectDesign of experiments (DoE)pt_BR
dc.subjectResponse surface methodology (RSM)pt_BR
dc.subjectStatistical experimental designpt_BR
dc.subject.wosStatistical Experimental-designpt_BR
dc.subject.wosActivating Protein-productionpt_BR
dc.subject.wosCell-density Culturept_BR
dc.subject.wosIntranasal Immunizationpt_BR
dc.subject.wosProtective Immunitypt_BR
dc.subject.wosPhytase Productionpt_BR
dc.subject.wosFusion Proteinpt_BR
dc.description.abstractPsaA, 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
dc.relation.ispartofJournal Of Industrial Microbiology & Biotechnologypt_BR
dc.relation.ispartofabbreviationJ. Ind. Microbiol. Biotechnol.pt_BR
dc.publisherSpringer Heidelbergpt_BR
dc.identifier.citationJournal Of Industrial Microbiology & Biotechnology. Springer Heidelberg, v. 39, n. 6, n. 897, n. 908, 2012.pt_BR
dc.sourceWeb of Sciencept_BR
dc.description.sponsorshipFundacao Oswaldo Cruz (Fiocruz)pt_BR
dc.description.provenanceMade available in DSpace on 2014-08-01T18:37:47Z (GMT). No. of bitstreams: 0 Previous issue date: 2012en
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