title: High-yield production of biologically active recombinant protein in shake flask culture by combination of enzyme-based glucose delivery and increased oxygen transfer.
abstract: This report describes the combined use of an enzyme-based glucose release system (EnBase®) and high-aeration shake flask (Ultra Yield Flask™). The benefit of this combination is demonstrated by over 100-fold improvement in the active yield of recombinant alcohol dehydrogenase expressed in E. coli. Compared to Terrific Broth and ZYM-5052 autoinduction medium, the EnBase system improved yield mainly through increased productivity per cell. Four-fold increase in oxygen transfer by the Ultra Yield Flask contributed to higher cell density with EnBase but not with the other tested media, and consequently the product yield per ml of EnBase culture was further improved.
authors.affiliation: BioSilta Oy, FI-90014 Oulu, Finland. kaisa.ukkonen@oulu.fi
authors.forename: Kaisa
authors.initials: K
authors.lastname: Ukkonen
authors.forename: Antti
authors.initials: A
authors.lastname: Vasala
authors.forename: Heikki
authors.initials: H
authors.lastname: Ojamo
authors.forename: Peter
authors.initials: P
authors.lastname: Neubauer
mesh_terms: D000426:Alcohol Dehydrogenase / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D061249:Batch Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D003470:Culture Media / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D005285:Fermentation; D005947:Glucose / Q000378:metabolism*; D007778:Lactobacillus / Q000201:enzymology; D010100:Oxygen / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D003470:Culture Media; D011994:Recombinant Proteins; D000426:Alcohol Dehydrogenase; D005947:Glucose; D010100:Oxygen