PMID:21698378
| Citation |
Li, Z, Kessler, W, van den Heuvel, J and Rinas, U (2011) Simple defined autoinduction medium for high-level recombinant protein production using T7-based Escherichia coli expression systems. Appl. Microbiol. Biotechnol. 91:1203-13 |
|---|---|
| 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. |
| Links |
PubMed Online version:10.1007/s00253-011-3407-z |
| Keywords |
Bacteriological Techniques/methods; Culture Media/chemistry; Escherichia coli/genetics; Escherichia coli/growth & development; Escherichia coli/metabolism; Gene Expression; Genetic Vectors; Podoviridae/genetics; Protein Engineering/methods; Recombinant Proteins/biosynthesis; Transcriptional Activation |
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