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In situ formation of H2O2 for P450 peroxygenases.

In situ formation of H2O2 for P450 peroxygenases.

Bioorganic & medicinal chemistry (2014-07-06)

Caroline E Paul, Ekaterina Churakova, Elmer Maurits, Marco Girhard, Vlada B Urlacher, Frank Hollmann

ABSTRACT

An in situ H2O2 generation approach to promote P450 peroxygenases catalysis was developed through the use of the nicotinamide cofactor analogue 1-benzyl-1,4-dihydronicotinamide (BNAH) and flavin mononucleotide (FMN). Final productivity could be enhanced due to higher enzyme stability at low H2O2 concentrations. The H2O2 generation represented the rate-limiting step, however it could be easily controlled by varying both FMN and BNAH concentrations. Further characterization can result in an optimized ratio of FMN/BNAH/O2/biocatalyst enabling high reaction rates while minimizing H2O2-related inactivation of the enzyme.

MATERIALS

Product Number

Brand

Product Description

Sigma-Aldrich

Ethyl acetate, ACS reagent, ≥99.5%

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Dodecanoic acid, 98%

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Dimethyl sulfoxide, meets EP testing specifications, meets USP testing specifications

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Dimethyl sulfoxide, Hybri-Max^™, sterile-filtered, BioReagent, suitable for hybridoma, ≥99.7%

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Sigma-Aldrich

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Dimethyl sulfoxide, European Pharmacopoeia (EP) Reference Standard

Sigma-Aldrich

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USP

Dimethyl sulfoxide, United States Pharmacopeia (USP) Reference Standard