Abstract
Laccases are multi-copper oxidoreductases (benzenediol:oxygen oxidoreductases, EC 1.10.3.2) able to oxidise a wide variety of phenolic and non-phenolic compounds. They are useful enzymes for a variety of applications, including bioremediation and craft pulp bio-bleaching as the most significant ones. There is a considerable interest to find new laccases through the exploration of biological diversity. Laccases have been found in plants, insects, and bacteria but predominantly in fungi: these enzymes have been documented in about 60 fungal strains. Microbial diversity constitutes a largely unexplored treasure chest with new laccases with a good potential for basic science and biotechnology. At present, due to our inability to cultivate most microbes, the only means of accessing the resources of the microbial world is to harvest genetic resources (“metagenomes”), which can further on be subjected to extensive screening programs. In this chapter, we provide an overview of screening methods to identify laccase-encoding genes from environmental resources.
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References
Thurston, C.F. (1994) The structure and function of fungal laccases. Microbiology 140, 19–26.
Sariaslani, F.S. (1989) Microbial enzymes for oxidation of organic molecules. Crit Rev Biotechnol 9, 171–257.
Bourbonnais, R. and Paice, M. (1990) Oxidation of non-phenolic substrates. An expanded role for laccase in lignin biodegradation. FEBS Lett 267, 99–102.
Eggert, C., Temp, U., Dean, J.F., and Eriksson, K.E. (1996) A fungal metabolite mediates degradation of non-phenolic lignin structures and synthetic lignin by laccase. FEBS Lett 391, 144–148.
Solomon, E.I., Sundaram, U.M., and Machonkin, T.E. (1996) Multicopper oxidases and oxygenases. Chem Rev 96, 2563–2605.
Camarero, S., Ibarra, D., Martínez, M.J., and Martínez, A.T. (2005) Lignin-derived compounds as efficient laccase mediators for decolorization of different types of recalcitrant dyes. Appl Environ Microbiol 71, 1775–1784.
Durán, N., Rosa, M.A., D’Annibale, A., and Gianfreda, L. (2002) Applications of laccases and tyrosinases (phenoloxidases) immobilized on different supports: a review. Enzyme Microb Tech 31, 907–931.
Wesenberg, D., Kyriakides, I., and Agathos, S.N. (2003) White-rot fungi and their enzymes for the treatment of industrial dye effluents. Biotechnol Adv 22, 161–187.
Arias, M.E., Arenas, M., Rodríguez, J., Soliveri, J., Ball, A.S., and Hernández, M. (2003) Kraft pulp biobleaching and mediated oxidation of a nonphenolic substrate by laccase from Streptomyces cyaneus CECT 3335. Appl Environ Microbiol 69, 1953–1958.
Bertrand, T., Jolivalt, C., Btiozzo, P., Caminade, E., Joly, N., Madzak, C., et al. (2002) Crystal structure of a four-copper laccase complexed with an arylamine: insights into substrate recognition and correlation with kinetics. Biochemistry 41, 7325–7333.
Gianfreda, L., Xu, F., and Bollag, J. (1999) Laccases: a useful group of oxidoreductive enzymes. Bioremediat J 3, 1–25.
Huttermann, A., Mai, C., and Kharazipour, A. (2001) Modification of lignin for the production of new compounded materials. Appl Microbiol Biotechnol 55, 387–394.
Mayer, A.M. and Staples, R.C. (2002) Laccase: new functions for an old enzyme. Phytochemistry 60, 551–565.
Parkinson, N.M., Conyers, C.M., Keen, J.N., MacNicoll, A.D., Smith, I., and Weaver, R.J. (2003) cDNAs encoding large venom proteins from the parasitoid Pimpla hypochondriaca was identified by random sequence analysis. Comp Biochem Physiol C-Toxicol Pharmacol 134, 513–520.
Enguita, F.J., Martins, L.O., Henriques, A.O., and Carrondo, M.A. (2003) Crystal structure of a bacterial endospore coat component – A laccase with enhanced thermostability properties. J Biol Chem 278, 19416–19425.
Alexandre, G. and Zhulin, I.B. (2000) Laccases are widspread in bacteria. Trends Biotechnol 18, 41–42.
Claus, H. (2003) Laccases and their occurrence in prokaryotes. Arch Microbiol 179, 145–150.
Claus, H. (2004) Laccases: structure, reactions, distribution. Micron 35, 93–96.
Vieites, J.M., Guazzaroni, M.E., Beloqui, A., Golyshin, P.N., and Ferrer, M. (2009) Metagenomics approaches in systems microbiology. FEMS Microbiol Rev 33, 236–255.
Beloqui, A., Pita, M., Yakimov, M.M., Polaina, J., Zumárraga, M., García-Arellano, H., et al. (2005) Novel polyphenol oxidase mined from a metagenome expresion library of bovine rumen: biochemical properties, structural analysis and phylogenetic relationships. J Biol Chem 281, 22933–22942.
Martins, L.O., Soares, C.M., Pereira, M.M., Teixeira, M., Costa, T., Jones, G.H., et al. (2002) Molecular and biochemical characterization of a highly stable bacterial laccase that occurs as a structural component of the Bacillus subtilis endospore coat. J Biol Chem 277, 18849–18859.
Ruijssenaars, H.J. and Hartmans, S. (2004) A cloned Bacillus halodurans multicopper oxidase exhibiting alkaline laccase activity. Appl Microbiol Biotechnol 65, 177–182.
Ducros, V., Brzozowski, A. M., Wilson, K. S., Brown, S. H., Ostergaard, P., Schneider, P., et al. (1998) Crystal structure of the type-2 Cu depleted laccase from Coprinus cinereus at 2.2 A resolution. Nature Struct Biol 5, 310–316.
Antorini, M., Herpoel-Gimbert, I., Choinowski, T., Sigoillot, J.C., Asther, M., Winterhalter, K., et al. (2002) Purification, crystallisation and X-ray diffraction study of fully functional laccases from two ligninolytic fungi. Biochim Biophys Acta 1594, 109–114.
Hakulinen, N., Kiiskinen, L.L., Kruus, K., Saloheimo, M., Paananen, A., Koivula, A., et al. (2002) Crystal structure of a laccase from Melanocarpus albomyces with an intact trinuclear copper site. Nature Struct Biol 9, 601–605.
McGuirl, M.A. and Dooley, D.M. (1999). Copper-containing oxidases. Curr Opin Chem Biol 3, 138–144.
Adler, E. and Eriksson, E. (1955) Guaiacylglycerol and its β-guaiacyl ether. Acta Chem Scand 9, 341–345.
Zumárraga, M., Bulter, T., Shleev, S., Polaina, J., Martínez-Arias, A., Plou, et al. (2007) In vitro evolution of a fungal laccase in high concentrations of organic cosolvents. Chem Biol 14, 1052–1064.
Ferrer, M., Beloqui, A., Vieites, J.M., Guazzaroni, M.E., Berger, I., and Aaroni, A. (2009) Interplay of metagenomics and in vitro compartmentalization. Microb Biotechnol 2, 31–39.
Ferrer, M., Beloqui, A., Timmis, K. N., and Golyshin P. N. (2009) Metagenomics for mining new genetic resources of microbial communities. J Mol Microbiol Biotechnol 16, 109–123.
Acknowledgements
This research was supported by the Spanish MEC BIO2006-11738, CSD2007-00005 and GEN2006-27750-C-4-E projects. A.B thanks the Spanish MEC for a FPU fellowship. P.N.G. was supported by Grant 0313751K from the Federal Ministry for Science and Education (BMBF) within the GenoMik-Plus initiative.
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Ferrer, M., Beloqui, A., Golyshin, P.N. (2010). Screening Metagenomic Libraries for Laccase Activities. In: Streit, W., Daniel, R. (eds) Metagenomics. Methods in Molecular Biology, vol 668. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-823-2_13
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DOI: https://doi.org/10.1007/978-1-60761-823-2_13
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