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- Title
Simple defined autoinduction medium for high-level recombinant protein production using T7-based Escherichia coli expression systems.
- Authors
Zhaopeng Li; Kessler, Wolfgang; Heuvel, Joop van den; 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 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 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.
- Publication
Applied Microbiology & Biotechnology, 2011, Vol 91, Issue 4, p1203
- ISSN
0175-7598
- Publication type
Academic Journal
- DOI
10.1007/s00253-011-3407-z