Simple defined autoinduction medium for high-level recombinant protein production using T7-based Escherichia coli expression systems.

2.50
Hdl Handle:
http://hdl.handle.net/10033/214650
Title:
Simple defined autoinduction medium for high-level recombinant protein production using T7-based Escherichia coli expression systems.
Authors:
Li, Zhaopeng; Kessler, Wolfgang; van den Heuvel, Joop; Rinas, Ursula
Abstract:
Protein production under the control of lac operon regulatory elements using autoinduction is based on diauxic growth of Escherichia coli on lactose after consumption of more preferred carbon substrates. A novel simple and cost-effective defined autoinduction medium using a mixture of glucose, glycerol, and lactose as carbon substrate and NH(4)(+) as sole nitrogen source without any supplementation of amino acids and vitamins was developed for T7-based E. coli expression systems. This medium was successfully employed in 96-well microtiter plates, test tubes, shake flasks, and 15-L bioreactor cultivations for production of different types of proteins achieving an average yield of 500 mg L(-1) product. Cell-specific protein concentrations and solubility were similar as during conventional isopropyl β-D-1-thiogalactopyranoside induction using Luria-Bertani broth. However, the final yield of target proteins was about four times higher, as a higher final biomass was achieved using this novel defined autoinduction broth.
Affiliation:
Helmholtz Centre for Infection Research (SB), Braunschweig, Germany.
Citation:
Simple defined autoinduction medium for high-level recombinant protein production using T7-based Escherichia coli expression systems. 2011, 91 (4):1203-13 Appl. Microbiol. Biotechnol.
Journal:
Applied microbiology and biotechnology
Issue Date:
Aug-2011
URI:
http://hdl.handle.net/10033/214650
DOI:
10.1007/s00253-011-3407-z
PubMed ID:
21698378
Type:
Article
Language:
en
ISSN:
1432-0614
Appears in Collections:
Publications of the RG Rekombinante Proteinexpression (RPEX)

Full metadata record

DC FieldValue Language
dc.contributor.authorLi, Zhaopengen
dc.contributor.authorKessler, Wolfgangen
dc.contributor.authorvan den Heuvel, Joopen
dc.contributor.authorRinas, Ursulaen
dc.date.accessioned2012-03-07T15:27:29Z-
dc.date.available2012-03-07T15:27:29Z-
dc.date.issued2011-08-
dc.identifier.citationSimple defined autoinduction medium for high-level recombinant protein production using T7-based Escherichia coli expression systems. 2011, 91 (4):1203-13 Appl. Microbiol. Biotechnol.en
dc.identifier.issn1432-0614-
dc.identifier.pmid21698378-
dc.identifier.doi10.1007/s00253-011-3407-z-
dc.identifier.urihttp://hdl.handle.net/10033/214650-
dc.description.abstractProtein production under the control of lac operon regulatory elements using autoinduction is based on diauxic growth of Escherichia coli on lactose after consumption of more preferred carbon substrates. A novel simple and cost-effective defined autoinduction medium using a mixture of glucose, glycerol, and lactose as carbon substrate and NH(4)(+) as sole nitrogen source without any supplementation of amino acids and vitamins was developed for T7-based E. coli expression systems. This medium was successfully employed in 96-well microtiter plates, test tubes, shake flasks, and 15-L bioreactor cultivations for production of different types of proteins achieving an average yield of 500 mg L(-1) product. Cell-specific protein concentrations and solubility were similar as during conventional isopropyl β-D-1-thiogalactopyranoside induction using Luria-Bertani broth. However, the final yield of target proteins was about four times higher, as a higher final biomass was achieved using this novel defined autoinduction broth.en
dc.language.isoenen
dc.subject.meshBacteriological Techniquesen
dc.subject.meshCulture Mediaen
dc.subject.meshEscherichia colien
dc.subject.meshGene Expressionen
dc.subject.meshGenetic Vectorsen
dc.subject.meshPodoviridaeen
dc.subject.meshProtein Engineeringen
dc.subject.meshRecombinant Proteinsen
dc.subject.meshTranscriptional Activationen
dc.titleSimple defined autoinduction medium for high-level recombinant protein production using T7-based Escherichia coli expression systems.en
dc.typeArticleen
dc.contributor.departmentHelmholtz Centre for Infection Research (SB), Braunschweig, Germany.en
dc.identifier.journalApplied microbiology and biotechnologyen
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