onr:"swepub:oai:DiVA.org:kth-25120"
Sökning: onr:"swepub:oai:DiVA.org:kth-25120" > Limiting factors in...
- 1 av 1
- Föregående post
- Nästa post
- Till träfflistan
Limiting factors in Escherichia coli fed-batch production of recombinant proteins
-
- Sandén, Anna Maria (författare)
- KTH,Bioteknologi
-
- Prytz, Ingela (författare)
- KTH,Bioteknologi
- Tubulekas, I. (författare)
- visa fler...
- Förberg, C. (författare)
- Le, H. (författare)
- Hektor, A. (författare)
- Neubauer, P. (författare)
- Pragai, Z. (författare)
- Harwood, C. (författare)
- Ward, A. (författare)
- Picon, A. (författare)
- de Mattos, J. T. (författare)
- Postma, P. (författare)
- Farewell, A. (författare)
- Nyström, T. (författare)
- Reeh, S. (författare)
- Pedersen, S. (författare)
- visa färre...
- (creator_code:org_t)
- 2002-11-22
- 2003
- Engelska.
- Ingår i: Biotechnology and Bioengineering. - : Wiley. - 0006-3592 .- 1097-0290. ; 81:2, s. 158-166
- Tidskriftsartikel (refereegranskat)
Abstract
Ämnesord
Stäng
- Fed-batch production of recombinant beta-galactosidase in E. coli was studied with respect to the specific growth rate at induction. The cultivations were designed to induce protein production by IPTG at a glucose feed rate corresponding to high (mu = 0.5 h(-1)) or low (mu = 0.1 h(-1)) specific growth rate. Protein production rate was approximately 100% higher at the higher specific growth rate, resulting in the accumulation of beta-galactosidase up to 30% of the total cell protein. Transcription analysis showed that beta-galactosidase-specific messenger RNA was immediately formed after induction (<5 min), but the amount was the same in both cases and was thus not the initial limiting factor. The content of ribosomes, as represented by rRNA, rapidly decreased with specific growth rate from a relative level of 100%, at the high specific growth rate, to 20% at the low specific growth rate. At high specific growth rate, ribosomes were additionally degraded upon induction due to the high production level. Translation therefore seemed to be the initial limiting factor of the protein synthesis capacity. The alarmone guanosine tetraphosphate increased at both high and low feed level inductions, indicating an induction-forced starvation of charged tRNA and/or glucose. The altered physiological status was also detected by the formation of acetic acid. However, the higher production rate resulted in high-level accumulation of acetic acid, which was absent at low feed rate production. Acetic acid production is thus coupled to the high product formation rate and is proposed to be due either to a precursor drain of Krebs cycle intermediates and a time lag before induction of the glyoxalate shunt, or to single amino acid overflow, since the model product is relatively poor in glycin and alanin. In conclusion, it is proposed that production at high specific growth rate becomes precursor-limited, while production at low specific growth rate is carbon- and/or energy-limited.
Ämnesord
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Nyckelord
- recombinant protein production
- fed-batch
- specific growth rate
- feed profile
- induction
- mRNA
- transcription
- translation
- acetic acid formation
- by-product formation
- ppGpp
- Biochemistry
- Biokemi
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
Hitta via bibliotek
- Biotechnology and Bioengineering (Sök värdpublikationen i LIBRIS)
Till lärosätets databas
- 1 av 1
- Föregående post
- Nästa post
- Till träfflistan
Hitta mer i SwePub
- Av författaren/redakt...
- Sandén, Anna Mar ...
- Prytz, Ingela
- Tubulekas, I.
- Förberg, C.
- Le, H.
- Hektor, A.
- visa fler...
- Neubauer, P.
- Pragai, Z.
- Harwood, C.
- Ward, A.
- Picon, A.
- de Mattos, J. T.
- Postma, P.
- Farewell, A.
- Nyström, T.
- Reeh, S.
- Pedersen, S.
- Larsson, Gen
- visa färre...
- Om ämnet
-
- NATURVETENSKAP
- NATURVETENSKAP
- och Biologi
- och Biokemi och mole ...
- Artiklar i publikationen
- Biotechnology an ...
- Av lärosätet
- Kungliga Tekniska Högskolan
Sök utanför SwePub
- Sök vidare i:
- Google Book Search
- Google Scholar
Kungliga biblioteket hanterar dina personuppgifter i enlighet med EU:s dataskyddsförordning (2018), GDPR. Läs mer om hur det funkar här.
Så här hanterar KB dina uppgifter vid användning av denna tjänst.
- Copyright © LIBRIS - Nationella bibliotekssystem
- LIBRIS.kb.se
