Isolation of two laccase genes from the white-rot fungus Pleurotus eryngii and heterologous expression of the pel3 encoded protein.
In this paper we report the cloning and nucleotide sequence analysis of two new laccase genes from the white-rot fungus Pleurotus eryngii, named pel3 and pel4. Comparison of the protein sequences deduced from these genes with laccases previously described in P. eryngii indicates that these genes codify for new laccases in this fungus. We described the expression of pel3 gene in two different Aspergillus niger strains. Both the laccase signal peptide and the glucoamylase preprosequence of A. niger were used to target the secretion of the active enzyme. The highest levels of laccase expression were obtained by combining the last construction with an A. niger strain deficient in extracellular proteases secretion. The characterization of catalytic properties of the recombinant enzyme, together with the setting-up of a heterologous expression system for pel3, will provide the basis to study the biotechnological applications of this enzyme.
Rodríguez
Enrique
E
Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, 28040 Madrid, Spain.
Ruiz-Dueñas
Francisco J
FJ
Kooistra
Rolf
R
Ram
Arthur
A
Martínez
Angel T
AT
Martínez
María Jesús
MJ
D000595:Amino Acid Sequence; D001234:Aspergillus niger / Q000201:enzymology / Q000235:genetics; D001483:Base Sequence; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D005087:Glucan 1,4-alpha-Glucosidase / Q000235:genetics / Q000378:metabolism; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D042845:Laccase / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D020076:Pleurotus / Q000235:genetics*; D016415:Sequence Alignment
D005656:Fungal Proteins; D007527:Isoenzymes; D042845:Laccase; D005087:Glucan 1,4-alpha-Glucosidase