title: The Pleurotus ostreatus laccase multi-gene family: isolation and heterologous expression of new family members.
abstract: This work was aimed at identifying and at characterizing new Pleurotus ostreatus laccases, in order to individuate the most suitable biocatalysts for specific applications. The existence of a laccase gene clustering was demonstrated in this basidiomycete fungus, and three new laccase genes were cloned, taking advantage of their closely related spatial organization on the fungus genome. cDNAs coding for two of the new laccases were isolated and expressed in the yeasts Saccharomyces cerevisiae and Kluyveromyces lactis, in order to optimize their production and to characterize the recombinant proteins. Analysis of the P. ostreatus laccase gene family allowed the identification of a "laccase subfamily" consisting of three genes. A peculiar intron-exon structure was revealed for the gene of one of the new laccases, along with a high instability of the recombinant enzyme due to lability of its copper ligand. This study allowed enlarging the assortment of P. ostreatus laccases and increasing knowledge to improve laccase production.
authors.affiliation: Department of Organic Chemistry and Biochemistry, University of Naples Federico II, Complesso Universitario Monte S. Angelo, via Cinthia, 4, 80126, Naples, Italy.
authors.forename: Cinzia
authors.initials: C
authors.lastname: Pezzella
authors.forename: Flavia
authors.initials: F
authors.lastname: Autore
authors.forename: Paola
authors.initials: P
authors.lastname: Giardina
authors.forename: Alessandra
authors.initials: A
authors.lastname: Piscitelli
authors.forename: Giovanni
authors.initials: G
authors.lastname: Sannia
authors.forename: Vincenza
authors.initials: V
authors.lastname: Faraco
mesh_terms: D000595:Amino Acid Sequence; D015139:Blotting, Southern; D022202:Chromosomes, Artificial, Bacterial; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D042845:Laccase / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D020076:Pleurotus / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
chemical_list: D018076:DNA, Complementary; D011994:Recombinant Proteins; D042845:Laccase