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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

Trizma^® base, ≥99.0% (T)

Sigma-Aldrich

Chlorotrimethylsilane, puriss., ≥99.0% (GC)

Supelco

Myristic acid, Pharmaceutical Secondary Standard; Certified Reference Material

Sigma-Aldrich

Trizma^® base, BioUltra, for molecular biology, ≥99.8% (T)

Sigma-Aldrich

Lauric acid, ≥98%, FCC, FG

Sigma-Aldrich

Myristic acid, natural, ≥98.5%, FG

Sigma-Aldrich

Diethyl ether

Sigma-Aldrich

Tris(hydroxymethyl)aminomethane, ACS reagent, ≥99.8%

Sigma-Aldrich

Myristic acid, ≥95%, FCC, FG

Sigma-Aldrich

Lauric acid, natural, ≥98%, FCC, FG

Sigma-Aldrich

Myristic acid, ≥98.0% (GC)

Sigma-Aldrich

Chlorotrimethylsilane, produced by Wacker Chemie AG, Burghausen, Germany, ≥99.0% (GC)

Supelco

Diethyl ether, analytical standard

Sigma-Aldrich

Myristic acid, Sigma Grade, ≥99%

Sigma-Aldrich

Trizma^® base, BioXtra, pH 10.5-12.0 (1 M in H₂O), ≥99.9% (titration)

Sigma-Aldrich

Trizma^® base, ≥99.9% (titration), crystalline

Sigma-Aldrich

Tromethamine, meets USP testing specifications

Supelco

Trizma^® base, reference material for titrimetry, certified by BAM, >99.5%

Supelco

Myristic acid, analytical standard

Sigma-Aldrich

Chlorotrimethylsilane, ≥98.0% (GC)

Sigma-Aldrich

Diethyl ether, suitable for HPLC, ≥99.9%, inhibitor-free

Sigma-Aldrich

Trizma^® base, BioPerformance Certified, meets EP, USP testing specifications, suitable for cell culture, ≥99.9% (titration)

Sigma-Aldrich

Chlorotrimethylsilane, purified by redistillation, ≥99%

Sigma-Aldrich

Trizma^® base, Primary Standard and Buffer, ≥99.9% (titration), crystalline

Sigma-Aldrich

Trizma^® base, puriss. p.a., ≥99.7% (T)

USP

Tromethamine, United States Pharmacopeia (USP) Reference Standard

Sigma-Aldrich

Diethyl ether, anhydrous, ACS reagent, ≥99.0%, contains BHT as inhibitor

Lauric acid, European Pharmacopoeia (EP) Reference Standard

Supelco

Tromethamine, pharmaceutical secondary standard, certified reference material

Trometamol, European Pharmacopoeia (EP) Reference Standard