Title: Characterization of periplasmic Escherichia coli protein expression at high cell densities
Abstract: ELECTROPHORESISVolume 20, Issue 4-5 p. 790-797 Research Article Characterization of periplasmic Escherichia coli protein expression at high cell densities Bo Franzén, Bo Franzén Astra Arcus AB, Preclinical R&D, Södertälje, SwedenSearch for more papers by this authorSusanne Becker, Corresponding Author Susanne Becker [email protected] Process R&D, Biopharma Bulk, Global Supply Europe, Pharmacia & Upjohn, Stockholm, SwedenProcess R&D, Biopharma Bulk, Global Supply Europe, Pharmacia & Upjohn, S-11287 Stockholm, Sweden, Fax: +46-8695-4146===Search for more papers by this authorRiitta Mikkola, Riitta Mikkola Process R&D, Biopharma Bulk, Global Supply Europe, Pharmacia & Upjohn, Stockholm, SwedenSearch for more papers by this authorKenneth Tidblad, Kenneth Tidblad Process R&D, Biopharma Bulk, Global Supply Europe, Pharmacia & Upjohn, Stockholm, SwedenSearch for more papers by this authorAgneta Tjernberg, Agneta Tjernberg Rockefeller University, New York, NY, USASearch for more papers by this authorStaffan Birnbaum, Staffan Birnbaum Process R&D, Biopharma Bulk, Global Supply Europe, Pharmacia & Upjohn, Stockholm, SwedenSearch for more papers by this author Bo Franzén, Bo Franzén Astra Arcus AB, Preclinical R&D, Södertälje, SwedenSearch for more papers by this authorSusanne Becker, Corresponding Author Susanne Becker [email protected] Process R&D, Biopharma Bulk, Global Supply Europe, Pharmacia & Upjohn, Stockholm, SwedenProcess R&D, Biopharma Bulk, Global Supply Europe, Pharmacia & Upjohn, S-11287 Stockholm, Sweden, Fax: +46-8695-4146===Search for more papers by this authorRiitta Mikkola, Riitta Mikkola Process R&D, Biopharma Bulk, Global Supply Europe, Pharmacia & Upjohn, Stockholm, SwedenSearch for more papers by this authorKenneth Tidblad, Kenneth Tidblad Process R&D, Biopharma Bulk, Global Supply Europe, Pharmacia & Upjohn, Stockholm, SwedenSearch for more papers by this authorAgneta Tjernberg, Agneta Tjernberg Rockefeller University, New York, NY, USASearch for more papers by this authorStaffan Birnbaum, Staffan Birnbaum Process R&D, Biopharma Bulk, Global Supply Europe, Pharmacia & Upjohn, Stockholm, SwedenSearch for more papers by this author First published: 27 April 1999 https://doi.org/10.1002/(SICI)1522-2683(19990101)20:4/5<790::AID-ELPS790>3.0.CO;2-ACitations: 13AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Abstract We have used two-dimensional electrophoresis (2-DE) to analyze changes in protein expression profiles during a microbial cultivation process on an industrial scale. An Escherichia coli strain W3110 containing the gene for recombinant human growth hormone production was used. Samples were taken at time intervals ranging from fast to slow growth rate (late growth phase at high cell density/starvation) and 2-DE analysis combined with image analysis using the PDQuest software showed significant alterations in expression levels of a number of proteins. Twenty-four protein spots were identified using a combination of matching with SWISS-2DPAGE E. coli map, N-terminal sequence analysis and mass spectrometry matrix-assisted laser desorption/ionization (MALDI). Two of the most abundant proteins expressed at late growth phase (pI 5.4/28 kDa and pI 5.5/28 kDa) were subjected to N-terminal sequence analysis after electrotransfer of the proteins from a preparative 2-DE gel to polyvinylidene difluoride (PVDF) membrane. Sequence tags of five amino acids in combination with approximate pI and Mr identified both proteins as deoxyribose phosphate aldolase (gene name deoC). In addition, both spots were subjected to tryptic in-gel digestion and analyzed using MALDI. Peptide mass fingerprints from both spots showed similar MALDI spectra and 10 of 10 tryptic fragments confirmed the identity as deoC. The identification of the acidic variant of deoC on 2-DE gels and the observation of this variant as induced during late growth phase is novel. Citing Literature Volume20, Issue4-51 January 1999Pages 790-797 RelatedInformation
Publication Year: 1999
Publication Date: 1999-01-01
Language: en
Type: article
Indexed In: ['crossref']
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