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)
Biological Research Centre (CIB)
Calle Ramiro de Maeztu 9, E-28040 Madrid, Spain
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publications
Total records: 113
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[ 2014 ] Fernandez-Fueyo E, Ruiz-Dueñas FJ, Martínez AT Engineering a fungal peroxidase that degrades lignin at very acidic pH Biotechnol. Biofuels, 7: 114
[ 2014 ] Fernandez-Fueyo E, Ruiz-Dueñas FJ, Martínez MJ, Romero A, Hammel KE, Medrano FJ, Martínez AT Ligninolytic peroxidase genes in the oyster mushroom genome: heterologous expression, molecular structure, catalytic and stability properties, and lignin-degrading ability Biotechnol. Biofuels, 7: 2
[ 2014 ] García-Ruiz E, Maté D, González-Pérez D, Molina-Espeja P, Camarero S, Martínez AT, Ballesteros A, Alcalde M Directed evolution of ligninolytic oxidoreductases: from functional expression to stabilization and beyond In "Cascade Biocatalysis. Integrating Stereoselective and Environmentally Friendly Reactions", First Edition. Edited by Sergio Riva and Wolf-Dieter Fessner. Wiley-VCH Verlag GmbH & Co
[ 2014 ] González-Pérez D, Alcalde M Assembly of evolved ligninolytic genes in Saccharomyces cerevisiae Bioengineered, 5: 254-263
[ 2014 ] González-Pérez D, García-Ruiz E, Ruiz-Dueñas FJ, Martínez AT, Alcalde M Structural determinants of oxidative stabilization in an evolved versatile peroxidase ACS-Catalysis, 4: 3891-3901
[ 2014 ] González-Pérez D, Molina-Espeja P, García-Ruiz E, Alcalde M Mutagenic Organized Recombination Process by Homologous In vivo Grouping (MORPHING) for directed enzyme evolution PlosOne, 9: 3
year2017
Mapping the Long-Range Electron Transfer Route in Ligninolytic Peroxidases
Acebes S, Ruiz-Dueñas FJ, Toubes M, Saez-Jimenez V, Pérez-Boada M, Lucas F, Martínez AT, Guallar V
J. Phys. Chem. B, 121: 3946-3954

Combining a computational analysis with site-directed mutagenesis, we have studied the long-range electron transfer pathway in versatile and lignin peroxidases, two enzymes of biotechnological interest that play a key role for fungal degradation of the bulky lignin molecule in plant biomass. The in silico study established two possible electron transfer routes starting at the surface tryptophan residue previously identified as responsible for oxidation of the bulky lignin polymer. Moreover, in both enzymes, a second buried tryptophan residue appears as a top electron transfer carrier, indicating the prevalence of one pathway. Site-directed mutagenesis of versatile peroxidase (from Pleurotus eryngii) allowed us to corroborate the computational analysis and the role played by the buried tryptophan (Trp244) and a neighbor phenylalanine residue (Phe198), together with the surface tryptophan, in the electron transfer. These three aromatic residues are highly conserved in all the sequences analyzed (up to a total of 169). The importance of the surface (Trp171) and buried (Trp251) tryptophan residues in lignin peroxidase has been also confirmed by directed mutagenesis of the Phanerochaete chrysosporium enzyme. Overall, the combined procedure identifies analogous electron transfer pathways in the long-range oxidation mechanism for both ligninolytic peroxidases, constituting a good example of how computational analysis avoids making extensive trial-error mutagenic experiments.

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