title: Separation of active laccases from Pleurotus sapidus culture supernatant using aqueous two-phase systems in centrifugal partition chromatography.
abstract: For the production of bio active compounds, e.g., active enzymes or antibodies, a conserved purification process with a minimum loss of active compounds is necessary. In centrifugal partition chromatography (CPC), the separation effect is based on the different distribution of the components to be separated between two immiscible liquid phases. Thereby, one liquid phase is kept stationary in chambers by a centrifugal field and the mobile phase is pumped through via connecting ducts. Aqueous two phase systems (ATPS) are known to provide benign conditions for biochemical products and seem to be promising when used in CPC for purification tasks. However, it is not known if active biochemical compounds can "survive" the conditions in a CPC where strong shear forces can occur due to the two-phasic flow under centrifugal forces. Therefore, this aspect has been faced within this study by the separation of active laccases from a fermentation broth of Pleurotus sapidus. After selecting a suitable ATPS and operating conditions, the activity yield was calculated and the preservation of the active enzymes could be observed. Therefore, CPC could be shown as potentially suitable for the purification of bio-active compounds.
authors: Laboratory of Plant and Process Design, Department of Biochemical and Chemical Engineering, TU Dortmund University, D-44227 Dortmund, Germany.
authors: C
authors: C
authors: Schwienheer
authors: Laboratory of Fluid Separations, Department of Biochemical and Chemical Engineering, TU Dortmund University, D-44227 Dortmund, Germany.
authors: A
authors: A
authors: Prinz
authors: Laboratory of Fluid Separations, Department of Biochemical and Chemical Engineering, TU Dortmund University, D-44227 Dortmund, Germany.
authors: T
authors: T
authors: Zeiner
authors: Laboratory of Plant and Process Design, Department of Biochemical and Chemical Engineering, TU Dortmund University, D-44227 Dortmund, Germany. Electronic address: juliane.merz@bci.tu-dortmund.de.
authors: J
authors: J
authors: Merz
mesh_terms: D002498:Centrifugation; D002853:Chromatography, Liquid / Q000379:methods*; D042845:Laccase / Q000302:isolation & purification*; D020076:Pleurotus / Q000201:enzymology*
chemical_list: D042845:Laccase
keywords: Activity preservation; Aqueous two-phase systems (ATPS); Centrifugal partition chromatography (CPC); Downstream processing; Laccase from Pleurotus sapidus