title: Characterization and structural analysis of the laccase I gene from the newly isolated ligninolytic basidiomycete PM1 (CECT 2971).
abstract: We have isolated and characterized the cDNA and genomic DNA coding for a phenoloxidase, laccase I, previously purified from culture supernatant of the newly isolated ligninolytic basidiomycete PM1 (CECT 2971). A cDNA library from basidiomycete PM1 was constructed, and laccase-encoding cDNAs were identified by screening with antiserum raised against the purified enzyme. The lac1 gene coding for the laccase was identified in a partial genomic library by using the isolated cDNA as a probe. Nucleotide sequence determination of the full-length cDNA revealed an open reading frame of 1,551 bp encoding a polypeptide of 517 amino acid residues with a putative signal peptide of 21 amino acid residues. Ten small introns interrupted the genomic DNA. A single 1.8-kb transcript mRNA was detected by Northern (RNA) blot analysis, and its 5' end maps to a position 51 bp upstream from the site of initiation of protein synthesis. Eukaryotic regulatory sequences, CAAT and TATA, were observed in the 5' flanking region, which also contains sequences similar to those of copper-regulated proteins. Comparative analysis of the predicted amino acid sequence showed that basidiomycete PM1 laccase I had great similarity to the laccases from Coriolus versicolor, Coriolus hirsutus, and Phlebia radiata.
authors.affiliation: Instituto de Microbiología-Bioquímica, Facultad de Biología, Consejo Superior de Investigaciones Científicas and Universidad de Salamanca, Spain.
authors.forename: P M
authors.initials: PM
authors.lastname: Coll
authors.forename: C
authors.initials: C
authors.lastname: Tabernero
authors.forename: R
authors.initials: R
authors.lastname: Santamaría
authors.forename: P
authors.initials: P
authors.lastname: Pérez
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001487:Basidiomycota / Q000201:enzymology* / Q000235:genetics; D002874:Chromosome Mapping; D005800:Genes, Fungal / Q000235:genetics*; D042845:Laccase; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
chemical_list: D010088:Oxidoreductases; D042845:Laccase