INGEBI   02650
INSTITUTO DE INVESTIGACIONES EN INGENIERIA GENETICA Y BIOLOGIA MOLECULAR "DR. HECTOR N TORRES"
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Cloning and expression of the laccases from the basidiomycete Trametes trogii BAFC 463 in Pichia pastoris
Autor/es:
CAMPOS PAULA; LEVIN LAURA; FORCHIASSIN FLAVIA; MENTABERRY ALEJANDRO; WIRTH SONIA
Lugar:
Potrero de Funes
Reunión:
Congreso; XLVII Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2011
Institución organizadora:
Sociedad Argentina de Investigación en Bioquímica y Biología Molecular
Resumen:
Fungal laccases are polyphenoloxidases that show many industrial applications as in delignification, textile dye or stain decolorization, water or soil bioremediation. Lignin degradation activity in basidiomycetes is due to the combined action of different enzymes, including laccases, Mn peroxidases, and lignin peroxidases. The white-rot fungus Trametes trogii BAFC 463 has shown an outstanding laccase activity (148,6 U/ml) and dye bleaching ability. This activity is due to at least 2 different isoenzymes: lcc1 and lcc2. In order to clone the lcc1 and lcc2 genes and look for other potential laccase isoenzymes in T. trogii, we design degenerated oligonucleotides corresponding to the conserved copper binding regions. Four different laccase coding genes were amplified from total cDNA, and complete coding sequence obtained by 5´RACE and 3´RACE techniques. Two of these cDNAs correspond to the lcc1 and lcc2 genes previously reported for T. trogii 201 (GenBank CAC13040 and CAL23367), and the other 2 correspond to novel isoenzymes (named lcc3 and lcc4), related to lcc1 from Coriolopsis gallica (97% similarity) and lcc2 from Trametes pubescens(86% similarity), respectively. All the four laccase coding genes were expressed in the methylotrophic yeast Pichia pastoris in order to obtain a simple source of laccase and to perform the biochemical and kinetic characterization for each isoenzyme.