Ylamide gel electrophoresis; T50: The temperature at which the enzyme loses 50 of its initial activity following incubation for 10 minutes. Competing interests The authors declare that they have no competing interests. Authors’ contributions DMM carried out the production and purification of ChU-B mutant both in P. pastoris and S. cerevisiae. DGP collaborated inside the purification of ChU-B mutant expressed in S. cerevisiae. DMM and DGP did the biochemical characterization of ChU-B produced in each yeasts. RK performed the vector constructions for laccase expression in P. pastoris. DMM wrote the very first draft on the manuscript. RL co-supervised the project and revised the manuscript. MA coordinated the project, supervised its development and wrote the final manuscript, which was read and approved by all authors. Acknowledgements This study is based upon a function funded by European Union Projects (grant numbers NMP4-SL-2009-229255-3D-Nanobiodevice, FP7-KBBE-20104-26537-Peroxicats and Expense Action CM0701) plus a Spanish National Project (Evofacel, BIO2010-19697) and by the Austrian Science Fund (FWF, P25148-B20). We thank Prof. S. Shleev from MalmUniversity (Sweden) for carrying out the measurements from the laccase activity in human plasma and blood. D.M.M. is grateful towards the CSIC for any JAE fellowship. Received: 8 January 2013 Accepted: 24 April 2013 Published: 30 April 2013 References 1. Baldrian P: Fungal laccases-occurrence and properties. FEMS Microbiol Rev 2006, 30:21542. two. Singh G, Bhalla A, Kaur P, Capalash N, Sharma P: Laccase from prokaryotes: A new source for an old enzyme. Rev Environ Sci Biotechnol 2011, 10:30926. 3. Laufer Z, Beckett RP, Minibayeva FV, Luthje S, Bottger M: Diversity of laccases from lichens in suborder Peltigerineae. Bryologist 2009, 112:41826. four. Davies GJ, Ducros V: Laccase. In Handbook of metalloproteins. Hoboken: John Wiley Sons, Ltd; 2006:1359368.Azadirachtin 5.Donanemab Ca s AI, Camarero S: Laccases and their natural mediators: biotechnological tools for sustainable eco-friendly processes.PMID:23399686 Biotechnol Adv 2010, 28:69405. six. Alcalde M: Laccases: biological functions, molecular structure and industrial applications. In Industrial enzymes. Structure, function and applications. Edited by Polaina J, MacCabe AP. Dordrecht: Springer Netherlands; 2007:46176. 7. Christenson A, Dimcheva N, Ferapontova EE, Gorton L, Ruzgas T, Stoica L, Shleev S, Yaropolov AL, Haltrich D, Thorneley RNF, Aust SD: Direct electron transfer in between ligninolytic redox enzymes and electrodes. Electroanal 2004, 16:1074092. 8. Heller A: Miniature biofuel cells. Phys Chem Chem Phys 2004, 6:20916. 9. Xu F: Applications of oxidoreductases: recent progress. Ind Biotechnol 2005, 1:380. 10. Rodgers CJ, Blanford CF, Giddens SR, Skamnioti P, Armstrong FA, Gurr SJ: Designer laccases: a vogue for high-potential fungal enzymes Trends Biotechnol 2010, 28:632. 11. Piscitelli A, Pezzella C, Giardina P, Faraco V, Giovanni S: Heterologous laccase production and its part in industrial applications. Bioengineered 2010, 1:25262. 12. MatD, Garc -Ruiz E, Camarero S, Alcalde M: Directed evolution of fungal laccases. Curr Genomics 2011, 12:11322. 13. Romanos MA, Scorer CA, Clare JJ: Foreign gene expression in yeast – a review. Yeast 1992, 8:42388. 14. Cobb RE, Si T, Zhao H: Directed evolution: an evolving and enabling synthetic biology tool. Curr Opi Chem Biol 2012, 16:28591. 15. Gonzalez-Perez D, Garcia-Ruiz E, Alcalde M: Saccharomyces cerevisiae in directed evolution: an efficient tool t.