Lyophilization conditions for the storage of monooxygenases

Hugo L. van Beek, Nina Beyer, Dick B. Janssen, Marco W. Fraaije*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

9 Citations (Scopus)

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 degrees 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 degrees C. (C) 2015 Elsevier B.V. All rights reserved.

Original languageEnglish
Pages (from-to)41-44
Number of pages4
JournalJournal of Biotechnology
Volume203
DOIs
Publication statusPublished - 10-Jun-2015

Keywords

  • Biocatalyst
  • Cryoprotection
  • Freeze-drying
  • Lyophilization
  • Monooxygenase
  • Sucrose
  • BAEYER-VILLIGER-MONOOXYGENASES
  • CYCLOHEXANONE MONOOXYGENASE
  • CYCLOPENTADECANONE MONOOXYGENASE
  • ORGANIC-SOLVENTS
  • STABILITY
  • PROTEIN
  • DISCOVERY
  • SUGAR
  • BIOCATALYSTS
  • OXIDATIONS

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