title: Lyophilization conditions for the storage of monooxygenases.
abstract: Cyclohexanone monooxygenase (CHMO) was used as a model enzyme to find suitable freeze-drying conditions for long-term storage of an isolated monooxygenase. CHMO is a Baeyer-Villiger monooxygenase (BVMO) known for its ability to catalyze a large number of oxidation reactions. With a focus on establishing the optimal formulation, additives were tested for enzyme stabilization during and after lyophilization. The results were successfully transferred to two other monooxygenases, namely the BVMO cyclopentadecanone monooxygenase (CPDMO) and a cytochrome P450 monooxygenase, P450 BM3. In the absence of a lyoprotectant, lyophilized P450 BM3 is almost completely inactivated, while the lyophilized BVMOs quickly lost activity when stored at 50°C. Lyophilization in the presence of 2% (w/v) sucrose was found to be the best formulation to preserve activity and protect against inactivation when stored as lyophilizate at 50°C.
authors: Molecular Enzymology Group, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands.
authors: Hugo L
authors: HL
authors: van Beek
authors: Molecular Enzymology Group, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands.
authors: Nina
authors: N
authors: Beyer
authors: Molecular Enzymology Group, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands.
authors: Dick B
authors: DB
authors: Janssen
authors: Molecular Enzymology Group, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands. Electronic address: M.W.Fraaije@rug.nl.
authors: Marco W
authors: MW
authors: Fraaije
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry*; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry*; D004356:Drug Storage; D005612:Freeze Drying; D009251:NADPH-Ferrihemoprotein Reductase / Q000737:chemistry*; D010105:Oxygenases / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D003577:Cytochrome P-450 Enzyme System; D010105:Oxygenases; C019961:cyclohexanone oxygenase; D009251:NADPH-Ferrihemoprotein Reductase; C052423:flavocytochrome P450 BM3 monoxygenases
keywords: Biocatalyst; Cryoprotection; Freeze-drying; Lyophilization; Monooxygenase; Sucrose