Switch in Cofactor Specificity of a Baeyer-Villiger Monooxygenase

Andy Beier, Sven Bordewick, Maika Genz, Sandy Schmidt, Tom Van Den Bergh, Christin Peters, Henk-Jan Joosten, Uwe T. Bornscheuer

Research output: Contribution to journalArticlepeer-review


Baeyer-Villiger monooxygenases (BVMOs) catalyze the oxidation of ketones to esters or lactones by using molecular oxygen and a cofactor. Type I BVMOs display a strong preference for NADPH. However, for industrial purposes NADH is the preferred cofactor, as it is ten times cheaper and more stable. Thus, we created a variant of the cyclohexanone monooxygenase from Acinetobacter sp. NCIMB 9871 (CHMOAcineto ); this used NADH 4200-fold better than NADPH. By combining structure analysis, sequence alignment, and literature data, 21 residues in proximity of the cofactor were identified and targeted for mutagenesis. Two combinatorial variants bearing three or four mutations showed higher conversions of cyclohexanone with NADH (79 %) compared to NADPH (58 %) as well as specificity. The structural reasons for this switch in cofactor specificity of a type I BVMO are especially a hydrogen-bond network coordinating the two hydroxy groups of NADH through direct interactions and bridging water molecules.

Original languageEnglish
Pages (from-to)2312-2315
Number of pages4
Issue number24
Publication statusPublished - 14 Dec 2016
Externally publishedYes


  • Acinetobacter
  • Binding Sites
  • Biocatalysis
  • Kinetics
  • Mixed Function Oxygenases
  • Molecular Dynamics Simulation
  • Mutagenesis, Site-Directed
  • NADP
  • Oxygenases
  • Protein Engineering
  • Protein Structure, Tertiary
  • Recombinant Proteins
  • Substrate Specificity
  • Journal Article

Cite this