The gene, protein and glycan structures of laccase from Pleurotus ostreatus.
A member of the laccase multigene family in Pleurotus ostreatus has been cloned and sequenced. The gene structure has been determined by comparison with the corresponding cDNA, synthesized by reverse transcription/PCR amplification. The gene encode a laccase isoenzyme of 533 amino acids which has already been purified and characterized [Palmieri, G., Giardina, P., Marzullo, L., Desiderio, B., Nitti, G., Cannio, R. & Sannia, G.(1993) Appl. Microbiol. Biotechnol. 39, 632-636]. More than 92% of the protein sequence, including the N and C termini, has been verified by fast-atom-bombardment mass spectrometry, thus confirming the correspondence between the gene and its protein product. The protein was N-glycosylated Asn444. Glycan analysis showed the presence of only a high-mannose structure containing varying numbers of mannose residues. The presence of O-linked oligosaccharides as well as other post-translational modification could be ruled out by the mass analysis.
Giardina
P
P
Dipartimento di Chimica Organica e Biologica, Università di Napoli Federico 11, Italy.
Aurilia
V
V
Cannio
R
R
Marzullo
L
L
Amoresano
A
A
Siciliano
R
R
Pucci
P
P
Sannia
G
G
D000595:Amino Acid Sequence; D001483:Base Sequence; D002236:Carbohydrate Conformation; D003001:Cloning, Molecular; D018076:DNA, Complementary; D006027:Glycosides / Q000737:chemistry; D006031:Glycosylation; D006868:Hydrolysis; D042845:Laccase; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D011124:Polyporaceae / Q000201:enzymology*; D011134:Polysaccharides / Q000737:chemistry*; D016339:Spectrometry, Mass, Fast Atom Bombardment
D018076:DNA, Complementary; D006027:Glycosides; D011134:Polysaccharides; D010088:Oxidoreductases; D042845:Laccase