Simple defined autoinduction medium for high-level recombinant protein production using T7-based Escherichia coli expression systems
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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.
KeywordsEscherichia coli Recombinant protein production Autoinduction Defined medium
Special thanks are given to Wolf-Dieter Schubert, Torsten Lührs, and Konrad Büssow for providing recombinant E. coli strains for testing general medium applicability. Partial support of the FORSYS Partner Project “Dynamics and regulation of the metabolic balance in Escherichia coli” is greatly acknowledged. We also thank Antonio Villaverde for “smart” suggestions.
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