Optimized oxidoreductases for medium and large scale industrial biotransformations
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Dr Marta Pérez-Boada
E-mail: MPBoada@cib.csic.es
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publications
Total records: 126
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[ 2014 ] Fernandez-Fueyo E, Castanera ER, Ruiz-Dueñas FJ, López-Lucendo MF, Ramírez L, Pisabarro AG, Martínez AT Ligninolytic peroxidase gene expression by Pleurotus ostreatus: Differential regulation in lignocellulose medium and effect of temperature and pH Fungal Gen. Biol., doi: 10.1016/j.fgb.2014.02.003
[ 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
year2014
Ligninolytic peroxidase genes in the oyster mushroom genome: heterologous expression, molecular structure, catalytic and stability properties, and lignin-degrading ability
Fernandez-Fueyo E, Ruiz-Dueñas FJ, Martínez MJ, Romero A, Hammel KE, Medrano FJ, Martínez AT
Biotechnol. Biofuels, 7: 2

Background
The genome of Pleurotus ostreatus, an important edible mushroom and a model ligninolytic organism of interest in lignocellulose biorefineries due to its ability to delignify agricultural wastes, was sequenced with the purpose of identifying and characterizing the enzymes responsible for lignin degradation.

Results
Heterologous expression of the class II peroxidase genes, followed by kinetic studies, enabled their functional classification. The resulting inventory revealed the absence of lignin peroxidases (LiPs) and the presence of three versatile peroxidases (VPs) and six manganese peroxidases (MnPs), the crystal structures of two of them (VP1 and MnP4) were solved at 1.0 to 1.1 Å showing significant structural differences. Gene expansion supports the importance of both peroxidase types in the white-rot lifestyle of this fungus. Using a lignin model dimer and synthetic lignin, we showed that VP is able to degrade lignin. Moreover, the dual Mn-mediated and Mn-independent activity of P. ostreatus MnPs justifies their inclusion in a new peroxidase subfamily. The availability of the whole POD repertoire enabled investigation, at a biochemical level, of the existence of duplicated genes. Differences between isoenzymes are not limited to their kinetic constants. Surprising differences in their activity T50 and residual activity at both acidic and alkaline pH were observed. Directed mutagenesis and spectroscopic/structural information were combined to explain the catalytic and stability properties of the most interesting isoenzymes, and their evolutionary history was analyzed in the context of over 200 basidiomycete peroxidase sequences.

Conclusions
The analysis of the P. ostreatus genome shows a lignin-degrading system where the role generally played by LiP has been assumed by VP. Moreover, it enabled the first characterization of the complete set of peroxidase isoenzymes in a basidiomycete, revealing strong differences in stability properties and providing enzymes of biotechnological interest.

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