title: An integrated scale-down plant for optimal recombinant enzyme production by Pichia pastoris.
abstract: An integrated bioprocess was created in a scale-down production plant by developing a two-stage enzyme production process with Pichia pastoris, containing a cell-breeding reactor and a production reactor in combination with a three-stage downstream process. To harvest the secreted enzymes, a disc separator and a cross-flow microfiltration clear the broth from the cells. Purification with hydrophobic interaction chromatography removes other proteins, concentrates the product, and prepares the enzyme solution for lyophilization. Fully automated and broad observable multi-stage parallel process courses have been developed using industrial process control systems and at-line measurements for enzyme concentration and enzyme activity. Optimal process conditions were found by application of Design of Experiments (DoE) for the production process.
authors.affiliation: Research Center of Bioprocess Engineering and Analytical Techniques, HAW - Hamburg University of Applied Sciences, Hamburg, Germany.
authors.forename: Kai
authors.initials: K
authors.lastname: Loegering
authors.forename: Christian
authors.initials: C
authors.lastname: Mueller
authors.forename: Jan-Patrick
authors.initials: JP
authors.lastname: Voss
authors.forename: Claudia
authors.initials: C
authors.lastname: Wagenfuehrer
authors.forename: Daniel
authors.initials: D
authors.lastname: Zahn
authors.forename: Hans-Peter
authors.initials: HP
authors.lastname: Bertelsen
authors.forename: Ulrich
authors.initials: U
authors.lastname: Scheffler
authors.forename: Reiner
authors.initials: R
authors.lastname: Luttmann
mesh_terms: D019149:Bioreactors; D010843:Pichia / Q000201:enzymology*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins