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

ZUSAMMENFASSUNG

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.

MATERIALIEN

Produktnummer

Marke

Produktbeschreibung

D2650

Sigma-Aldrich

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

472301

Sigma-Aldrich

Dimethylsulfoxid, ACS reagent, ≥99.9%

D8418

Sigma-Aldrich

Dimethylsulfoxid, for molecular biology

319902

Sigma-Aldrich

Ethylacetat, ACS reagent, ≥99.5%

34869

Sigma-Aldrich

Dimethylsulfoxid, suitable for HPLC, ≥99.7%

D2438

Sigma-Aldrich

Dimethylsulfoxid, sterile-filtered, BioPerformance Certified, meets EP, USP testing specifications, suitable for hybridoma

T1503

Sigma-Aldrich

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

34858

Sigma-Aldrich

Ethylacetat, suitable for HPLC, ≥99.7%

673811

Sigma-Aldrich

Diethylether, anhydrous, ACS reagent, ≥99.0%, contains BHT as inhibitor

D5879

Sigma-Aldrich

Dimethylsulfoxid, ReagentPlus^®, ≥99.5%

309966

Sigma-Aldrich

Diethylether, suitable for HPLC, ≥99.9%, inhibitor-free

650528

Sigma-Aldrich

Ethylacetat, HPLC Plus, for HPLC, GC, and residue analysis, 99.9%

T6066

Sigma-Aldrich

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

346136

Sigma-Aldrich

Diethylether, ACS reagent, anhydrous, ≥99.0%, contains BHT as inhibitor

D4540

Sigma-Aldrich

Dimethylsulfoxid, ≥99.5% (GC), suitable for plant cell culture

41640

Sigma-Aldrich

Dimethylsulfoxid, puriss. p.a., ACS reagent, ≥99.9% (GC)

276855

Sigma-Aldrich

Dimethylsulfoxid, anhydrous, ≥99.9%

92361

Sigma-Aldrich

Chlorotrimethylsilan, ≥98.0% (GC)

T6687

Sigma-Aldrich

Tromethamin, meets USP testing specifications

T1378

Sigma-Aldrich

Sigma 7-9^®, ≥99% (titration), crystalline

439169

Sigma-Aldrich

Ethylacetat, suitable for HPLC, ≥99.8%

93362

Sigma-Aldrich

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

32203

Sigma-Aldrich

Diethylether, contains BHT as inhibitor, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.8% (GC)

386529

Sigma-Aldrich

Chlorotrimethylsilan, purified by redistillation, ≥99%

41639

Sigma-Aldrich

Dimethylsulfoxid, BioUltra, for molecular biology, ≥99.5% (GC)

93352

Sigma-Aldrich

Trizma^® Base, ≥99.0% (T)

33211

Sigma-Aldrich

Ethylacetat, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.5% (GC)

27227

Sigma-Aldrich

Ethylacetat, puriss., meets analytical specification of Ph. Eur., BP, NF, ≥99.5% (GC)

W261408

Sigma-Aldrich

Laurinsäure, ≥98%, FCC, FG

T6791

Sigma-Aldrich

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