title: Prediction of inclusion body solubilization from shaken to stirred reactors.
abstract: Inclusion bodies (IBs) were solubilized in a µ-scale system using shaking microtiter plates or a stirred tank reactor in a laboratory setting. Characteristic dimensionless numbers for mixing, the Phase number Ph and Reynolds number Re did not correlate with the kinetics and equilibrium of protein solubilization. The solubilization kinetics was independent of the mixing system, stirring or shaking rate, shaking diameter, and energy input. Good agreement was observed between the solubilization kinetics and yield on the µ-scale and laboratory setting. We show that the IB solubilization process is controlled predominantly by pore diffusion. Thus, for the process it is sufficient to keep the IBs homogeneously suspended, and additional power input will not improve the process. The high-throughput system developed on the µ-scale can predict solubilization in stirred reactors up to a factor of 500 and can therefore be used to determine optimal solubilization conditions on laboratory and industrial scale.
authors.affiliation: Austrian Centre of Industrial Biotechnology, Muthgasse 11, 1190, Vienna, Austria.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Walther
authors.forename: Sabrina
authors.initials: S
authors.lastname: Mayer
authors.forename: Alexandru
authors.initials: A
authors.lastname: Trefilov
authors.forename: Gerhard
authors.initials: G
authors.lastname: Sekot
authors.forename: Rainer
authors.initials: R
authors.lastname: Hahn
authors.forename: Alois
authors.initials: A
authors.lastname: Jungbauer
authors.forename: Astrid
authors.initials: A
authors.lastname: Dürauer
mesh_terms: D019149:Bioreactors / Q000382:microbiology*; D001709:Biotechnology / Q000295:instrumentation / Q000379:methods; D004926:Escherichia coli / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods; D002479:Inclusion Bodies / Q000737:chemistry* / Q000378:metabolism*; D007700:Kinetics; D012995:Solubility
keywords: E. coli; microtiter plate; mixing techniques; scale up; solubilization; stirred tank reactor