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Titolo:
COMPARISON OF TEMPERATURE-INDUCED AND ISOPROPYL-BETA-D-THIOGALACTO-PYRANOSIDE-INDUCED SYNTHESIS OF BASIC FIBROBLAST GROWTH-FACTOR IN HIGH-CELL-DENSITY CULTURES OF RECOMBINANT ESCHERICHIA-COLI
Autore:
SEEGER A; SCHNEPPE B; MCCARTHY JEG; DECKWER WD; RINAS U;
Indirizzi:
GBF NATL RES CTR BIOTECHNOL,DIV BIOCHEM ENGN,MASCHERODER WEG 1 D-38124 BRAUNSCHWEIG GERMANY GBF NATL RES CTR BIOTECHNOL,DIV BIOCHEM ENGN D-38124 BRAUNSCHWEIG GERMANY DEPT GENE EXPRESS BRAUNSCHWEIG GERMANY
Titolo Testata:
Enzyme and microbial technology
fascicolo: 10, volume: 17, anno: 1995,
pagine: 947 - 953
SICI:
0141-0229(1995)17:10<947:COTAI>2.0.ZU;2-4
Fonte:
ISI
Lingua:
ENG
Soggetto:
FED-BATCH CULTURES; CULTIVATION; EXPRESSION; PROTEINS; SYSTEM; GENES; GALACTOSIDASE;
Keywords:
BASIC FIBROBLAST GROWTH FACTOR; COMPARISON OF DIFFERENT EXPRESSION SYSTEMS; HIGH-CELL-DENSITY CULTIVATION; RECOMBINANT ESCHERICHIA COIL;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
23
Recensione:
Indirizzi per estratti:
Citazione:
A. Seeger et al., "COMPARISON OF TEMPERATURE-INDUCED AND ISOPROPYL-BETA-D-THIOGALACTO-PYRANOSIDE-INDUCED SYNTHESIS OF BASIC FIBROBLAST GROWTH-FACTOR IN HIGH-CELL-DENSITY CULTURES OF RECOMBINANT ESCHERICHIA-COLI", Enzyme and microbial technology, 17(10), 1995, pp. 947-953

Abstract

Two different expression systems were developed for expression of thecDNA encoding human basic fibroblast growth factor (bFGF) using Escherichia coli TG1 as host organism. The bFGF structural gene was cloned into two vectors differing only with respect to the promoter, which was either the bacteriophage lambda PRPL- or the E. coli lac-promoter. The resulting expression systems were studied in high-cell density cultures. Cells were grown in a fed-batch procedure with a predetermined exponential feeding rate based on mass balances and kinetic equations to ensure constant specific growth rates. Prior to induction, cells were grown at 30 degrees C. Product formation was induced by either a temperature shift from 30 to 42 degrees C or by the addition of isopropyl-beta-D-thiogalactopyranoside (IPTG). Under comparable culture conditions-induction of bFGF expression at 41 to 45 g l(-1) dry cell weight and the addition of feed medium with identical rates-bFGF accumulated to 4.9 and 1.1 g l(-1) using the temperature- and IPTG-inducible expression systems, respectively. The final biomass concentrations obtained using temperature- and IPTG-inducible expression systems were 61 and 135 g l(-1) dry cell weight, whereas the specific product concentrations were 80 and 8.1 mg bFGF g(-1) dry cell weight, respectively. When a temperature shift was used for product induction, 30% of bFGF was recovered as inclusion bodies in the insoluble cell fraction. IPTG-dependent induction yielded exclusively soluble bFGF.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 18/09/20 alle ore 17:04:59