title: Glucose-limited high cell density cultivations from small to pilot plant scale using an enzyme-controlled glucose delivery system.
abstract: The enzyme controlled substrate delivery cultivation technology EnBase(®) Flo allows a fed-batch-like growth in batch cultures. It has been previously shown that this technology can be applied in small cultivation vessels such as micro- and deep well plates and also shake flasks. In these scales high cell densities and improved protein production for Escherichia coli cultures were demonstrated. This current study aims to evaluate the scalability of the controlled glucose release technique to pilot scale bioreactors. Throughout all scales, that is, deep well plates, 3 L bioreactor and 150 L bioreactor cultivations, the growth was very similar and the model protein, a recombinant alcohol dehydrogenase (ADH) was produced with a high yield in soluble form. Moreover, EnBase Flo also was successfully used as a controlled starter culture in high cell density fed-batch cultivations with external glucose feeding. Here the external feeding pump was started after overnight cultivation with EnBase Flo. Final optical densities in these cultivations reached 120 (corresponding to about 40 g L(-1) dry cell weight) and a high expression level of ADH was obtained. The EnBase cultivation technology ensures a controlled initial cultivation under fed-batch mode without the need for a feeding pump. Because of the linear cell growth under glucose limitation it provides optimal and robust starting conditions for traditional external feed-based processes.
authors.affiliation: Laboratory of Bioprocess Engineering, Department of Biotechnology, Technische Universität Berlin, Ackerstraße 71-76, ACK-24, D-13355 Berlin, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Glazyrina
authors.forename: Mirja
authors.initials: M
authors.lastname: Krause
authors.forename: Stefan
authors.initials: S
authors.lastname: Junne
authors.forename: Florian
authors.initials: F
authors.lastname: Glauche
authors.forename: Dirk
authors.initials: D
authors.lastname: Storm
authors.forename: Dirk
authors.initials: D
authors.lastname: Strom
authors.forename: Peter
authors.initials: P
authors.lastname: Neubauer
mesh_terms: D061249:Batch Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D019149:Bioreactors / Q000382:microbiology*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D017022:Flow Injection Analysis / Q000295:instrumentation* / Q000379:methods; D005947:Glucose / Q000737:chemistry / Q000378:metabolism*; D005949:Glucose Oxidase / Q000378:metabolism*; D010865:Pilot Projects
chemical_list: D005949:Glucose Oxidase; D005947:Glucose