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Simple defined autoinduction medium for high-level recombinant protein production using T7-based Escherichia coli expression systems.
Li, Zhaopeng Kessler, Wolfgang van den Heuvel, Joop Rinas, Ursula
Li, Zhaopeng
Kessler, Wolfgang
van den Heuvel, Joop
Rinas, Ursula
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2011-08
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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.
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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.
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Article
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en
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1432-0614