title: Recombinant expression of Pleurotus ostreatus laccases in Kluyveromyces lactis and Saccharomyces cerevisiae.
abstract: Heterologous expression of Pleurotus ostreatus POXC and POXA1b laccases in two yeasts, Kluyveromyces lactis and Saccharomyces cerevisiae, was performed. Both transformed hosts secreted recombinant active laccases, although K. lactis was much more effective than S. cerevisiae. rPOXA1b transformants always had higher secreted activity than rPOXC transformants did. The lower tendency of K. lactis with respect to S. cerevisiae to hyperglycosylate recombinant proteins was confirmed. Recombinant laccases from K. lactis were purified and characterised. Specific activities of native and recombinant POXA1b are similar. On the other hand, rPOXC specific activity is much lower than that of the native protein, perhaps due to incomplete or incorrect folding. Both recombinant laccase signal peptides were correctly cleaved, with rPOXA1b protein having two C-terminal amino acids removed. The availability of the established recombinant expression system provides better understanding of laccase structure-function relationships and allows the development of new oxidative catalysts through molecular evolution techniques.
authors: Dipartimento di Chimica Organica e Biochimica, Università di Napoli Federico II, Complesso Universitario Monte S. Angelo, via Cinthia, 80126 Naples, Italy. sannia@unina.it
authors: Alessandra
authors: A
authors: Piscitelli
authors: Paola
authors: P
authors: Giardina
authors: Cristina
authors: C
authors: Mazzoni
authors: Giovanni
authors: G
authors: Sannia
mesh_terms: D000595:Amino Acid Sequence; D001709:Biotechnology / Q000379:methods; D007716:Kluyveromyces / Q000201:enzymology* / Q000235:genetics; D042845:Laccase / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D020076:Pleurotus / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D042845:Laccase