Optimized oxidoreductases for medium and large scale industrial biotransformations
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Dr Marta Pérez-Boada
E-mail: MPBoada@cib.csic.es
Consejo Superior de Investigaciones Científicas (CSIC)
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publications
Total records: 126
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[ 2015 ] Viña-Gonzalez J, González-Pérez D, Ferreira P, Martínez AT, Alcalde M Focused directed evolution of aryl-alcohol oxidase in yeast using chimeric signal peptides Appl. Environ. Microbiol., 81: 6451-6462
[ 2015 ] Wang X, Ullrich R, Hofrichter M, Groves JT Heme-thiolate ferryl of aromatic peroxygenase is basic and reactive Proc. Natl. Acad. Sci. USA, 112: 3686-3691
[ 2014 ] Barrasa JM, Blanco MN, Esteve-Raventós F, Altés A, Checa J, Martínez AT, Ruiz-Dueñas FJ Wood and humus decay strategies by white-rot basidiomycetes correlate with two different dye decolorization and enzyme secretion patterns on agar plates Fungal Gen. Biol., doi: 10.1016/j.fgb.2014.03.007
[ 2014 ] Camarero S, Martínez MJ, Martínez AT Understanding lignin biodegradation for the improved utilization of plant biomass in modern biorefineries Biofuels, Bioprod. Bioref., 8: 615-625
[ 2014 ] Carro J, Ferreira P, Rodríguez L, Prieto A, Serrano A, Balcells B, Ardá A, Jiménez-Barbero J, Gutiérrez A, Ullrich R, Hofrichter M, Martínez AT 5-Hydroxymethylfurfural conversion by fungal aryl-alcohol oxidase and unspecific peroxygenase FEBS J., 282: 3218-3229
[ 2014 ] Fernandez-Fueyo E, Acebes S, Ruiz-Dueñas FJ, Martínez MJ, Romero A, Medrano FJ, Guallar V, Martínez AT Structural implications of the C-terminal tail in the catalytic and stability properties of manganese peroxidases from ligninolytic fungi Acta Crystal. D, 70: 3253-3265
year2014
Structural determinants of oxidative stabilization in an evolved versatile peroxidase
González-Pérez D, García-Ruiz E, Ruiz-Dueñas FJ, Martínez AT, Alcalde M
ACS-Catalysis, 4: 3891-3901

Versatile peroxidases (VP) are promiscuous biocatalysts with the highest fragility to hydroperoxides yet reported due to a complex molecular architecture, with three catalytic sites and several oxidation pathways. To improve the VP resistance to H2O2, an evolved version of this enzyme was subjected to a range of directed evolution and hybrid strategies in Saccharomyces cerevisiae. After five generations of random, saturation, and domain mutagenesis, together with in vivo DNA recombination, several structural determinants behind the oxidative destabilization of the enzyme were unmasked. To establish a balance between activity and stability, selected beneficial mutations were introduced into novel mutational environments by the in vivo exchange of sequence blocks, promoting epistatic interactions. The best variant of this process accumulated 8 mutations that increased the half-life of the protein from 3 (parental type) to 35 min in the presence of 3000 equiv of H2O2 and with a 6 °C upward shift in thermostability. Multiple structural alignment with other H2O2-tolerant heme peroxidases help to understand the possible roles played by the new mutations in the overall oxidative stabilization of these enzymes.

Official webpage of indox [ 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 garcíarincón