title: Simple defined autoinduction medium for high-level recombinant protein production using T7-based Escherichia coli expression systems.
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.
authors.affiliation: Helmholtz Centre for Infection Research (SB), Braunschweig, Germany.
authors.forename: Zhaopeng
authors.initials: Z
authors.lastname: Li
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Kessler
authors.forename: Joop
authors.initials: J
authors.lastname: van den Heuvel
authors.forename: Ursula
authors.initials: U
authors.lastname: Rinas
mesh_terms: D001431:Bacteriological Techniques / Q000379:methods; D003470:Culture Media / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D015870:Gene Expression; D005822:Genetic Vectors; D017902:Podoviridae / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D015533:Transcriptional Activation
chemical_list: D003470:Culture Media; D011994:Recombinant Proteins