Optimized oxidoreductases for medium and large scale industrial biotransformations
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126
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[ 2015 ]
Hofrichter M, Kellner H, Pecyna MJ, Ullrich R Fungal unspecific peroxygenases: heme-thiolate proteins that combine peroxidase and cytochrome p450 properties
Adv. Exp. Med. Biol., 851: 341-368
[ 2015 ]
Kracher D, Zahma K, Schulz C, Sygmund C, Gorton L, Ludwig R Inter-domain electron transfer in cellobiose dehydrogenase: modulation by pH and divalent cations
FEBS J., doi: 10.1111/febs.13310
[ 2015 ]
Linde D, Pogni R, Cañellas M, Lucas F, Guallar V, Baratto MC, Sinicropi A, Saez-Jimenez V, Coscolín C, Romero A, Medrano FJ, Ruiz-Dueñas FJ, Martínez AT Catalytic surface radical in dye-decolorizing peroxidase: A computational, spectroscopic and site-directed mutagenesis study
Biochem. J., 466: 253-262
[ 2015 ]
Linde D, Ruiz-Dueñas FJ, Fernandez-Fueyo E, Guallar V, Hammel KE, Pogni R, Martínez AT Basidiomycete DyPs: Genomic diversity, structural-functional aspects, reaction mechanism and environmental significance
Arch. Biochem. Biophys., 574: 66-74
[ 2015 ]
Maté D, Alcalde M Laccase engineering: From rational design to directed evolution
Biotechnol. Adv., 33: 25-40
[ 2015 ]
Molina-Espeja P, Ma S, Maté D, Ludwig R, Alcalde M Tandem-yeast expression system for engineering and producing unspecific peroxygenase
Enz. Microb. Technol., 73: 29-33
year2015
Engineering an enzymatic regeneration system for NAD(P)H oxidation
Pham NH, Hollmann F, Kracher D, Preims M, Haltrich D, Ludwig R
J. Mol. Cat. B, 120: 38-46
A recently proposed coenzyme regeneration system employing laccase and a number of various redox mediators for the oxidation of NAD(P)H was studied in detail by kinetic characterization of individual reaction steps. Reaction engineering by modeling was used to optimize the employed enzyme, coenzyme as well as redox mediator concentrations. Glucose dehydrogenase from Bacillus sp. served as a convenient model of synthetic enzymes that depend either on NAD+ or NADP+. The suitability of laccase from Trametes pubescens in combination with acetosyringone or syringaldazine as redox mediator was tested for the regeneration (oxidation) of both coenzymes. In a first step, pH profiles and catalytic constants of laccase for the redox mediators were determined. Then, second-order rate constants for the oxidation of NAD(P)H by the redox mediators were measured. In a third step, the rate equation for the entire enzymatic process was derived and used to build a MATLAB model. After verifying the agreement of predicted vs. experimental data, the model was used to calculate different scenarios employing varying concentrations of regeneration system components. The modeled processes were experimentally tested and the results compared to the predictions. It was found that the regeneration of NADH to its oxidized form was performed very efficiently, but that an excess of laccase activity leads to a high concentration of the oxidized form of the redox mediator – a phenoxy radical – which initiates coupling (dimerization or polymerization) and enzyme deactivation.
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[ industrialoxidoreductases ]. Optimized oxidoreductases for medium and large scale industrial biotransformations. This project has received funding from the European Union’s Seventh Framework Programme for research, technological development and demonstration under Grant Agreement nº: FP7-KBBE-2013-7-613549. © indox 2013. Developed by
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