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 journalArticleResearchpeer-review

Abstract

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
JournalChemBioChem
Volume17
Issue number24
DOIs
Publication statusPublished - 14 Dec 2016
Externally publishedYes

Fingerprint

Mixed Function Oxygenases
NAD
NADP
Switches
Bearings (structural)
Mutagenesis
Acinetobacter
Molecular oxygen
Sequence Alignment
Lactones
Ketones
Hydrogen
Hydrogen bonds
Esters
Oxygen
Oxidation
Mutation
Molecules
Water

Keywords

  • 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

Beier, A., Bordewick, S., Genz, M., Schmidt, S., Van Den Bergh, T., Peters, C., ... Bornscheuer, U. T. (2016). Switch in Cofactor Specificity of a Baeyer-Villiger Monooxygenase. ChemBioChem, 17(24), 2312-2315. https://doi.org/10.1002/cbic.201600484

Switch in Cofactor Specificity of a Baeyer-Villiger Monooxygenase. / Beier, Andy; Bordewick, Sven; Genz, Maika; Schmidt, Sandy; Van Den Bergh, Tom; Peters, Christin; Joosten, Henk-Jan; Bornscheuer, Uwe T.

In: ChemBioChem, Vol. 17, No. 24, 14.12.2016, p. 2312-2315.

Research output: Contribution to journalArticleResearchpeer-review

Beier, A, Bordewick, S, Genz, M, Schmidt, S, Van Den Bergh, T, Peters, C, Joosten, H-J & Bornscheuer, UT 2016, 'Switch in Cofactor Specificity of a Baeyer-Villiger Monooxygenase' ChemBioChem, vol. 17, no. 24, pp. 2312-2315. https://doi.org/10.1002/cbic.201600484
Beier A, Bordewick S, Genz M, Schmidt S, Van Den Bergh T, Peters C et al. Switch in Cofactor Specificity of a Baeyer-Villiger Monooxygenase. ChemBioChem. 2016 Dec 14;17(24):2312-2315. https://doi.org/10.1002/cbic.201600484
Beier, Andy ; Bordewick, Sven ; Genz, Maika ; Schmidt, Sandy ; Van Den Bergh, Tom ; Peters, Christin ; Joosten, Henk-Jan ; Bornscheuer, Uwe T. / Switch in Cofactor Specificity of a Baeyer-Villiger Monooxygenase. In: ChemBioChem. 2016 ; Vol. 17, No. 24. pp. 2312-2315.
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