title: Glycosylated yellow laccases of the basidiomycete Stropharia aeruginosa.
abstract: Here we describe the identification, purification and characterisation of glycosylated yellow laccase proteins from the basidiomycete fungus Stropharia aeruginosa. Biochemical characterisation of two yellow laccases, Yel1p and Yel3p, show that they are both secreted, monomeric, N-glycosylated proteins of molecular weight around 55kDa with substrate specificities typical of laccases, but lacking the absorption band at 612nm typical of the blue laccase proteins. Low coverage, high throughput 454 transcriptome sequencing in combination with inverse-PCR was used to identify cDNA sequences. One of the cDNA sequences has been assigned to the Yel1p protein on the basis of identity between the translated protein sequence and the peptide data from the purified protein, and the full length gene sequence has been obtained. Biochemical properties, substrate specificities and protein sequence data have been used to discuss the unusual spectroscopic properties of S. aeruginosa proteins in the context of recent theories about the differences between yellow and blue laccases.
authors.affiliation: Institute of Integrative Biology, University of Liverpool, Biosciences Building, Crown Street, Liverpool L69 7ZB, United Kingdom; School of Environment and Energy, Peking University Shenzhen Graduate School, Shenzhen 518055, China.
authors.forename: Maurycy
authors.initials: M
authors.lastname: Daroch
authors.affiliation: Institute of Integrative Biology, University of Liverpool, Biosciences Building, Crown Street, Liverpool L69 7ZB, United Kingdom.
authors.forename: Catharine A
authors.initials: CA
authors.lastname: Houghton
authors.affiliation: Institute of Integrative Biology, University of Liverpool, Biosciences Building, Crown Street, Liverpool L69 7ZB, United Kingdom.
authors.forename: Jonathan K
authors.initials: JK
authors.lastname: Moore
authors.affiliation: Institute of Integrative Biology, University of Liverpool, Biosciences Building, Crown Street, Liverpool L69 7ZB, United Kingdom.
authors.forename: Mark C
authors.initials: MC
authors.lastname: Wilkinson
authors.affiliation: Department of Chemistry, University of Liverpool, Crown Street, Liverpool L69 7ZD, United Kingdom.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Carnell
authors.affiliation: Institute of Integrative Biology, University of Liverpool, Biosciences Building, Crown Street, Liverpool L69 7ZB, United Kingdom.
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Bates
authors.affiliation: Institute of Ageing and Chronic Disease, Faculty of Health & Life Sciences, Department of Musculoskeletal Biology, University of Liverpool, Duncan Building, Daulby Street, Liverpool L69 3GA, United Kingdom. Electronic address: iwanejko@liverpool.ac.uk.
authors.forename: Lesley A
authors.initials: LA
authors.lastname: Iwanejko
mesh_terms: D000363:Agaricales / Q000201:enzymology* / Q000235:genetics; D000595:Amino Acid Sequence; D001483:Base Sequence; D002845:Chromatography; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D015723:Gene Library; D005800:Genes, Fungal; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D006031:Glycosylation; D042845:Laccase / Q000737:chemistry / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011499:Protein Processing, Post-Translational; D012331:RNA, Fungal / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013056:Spectrophotometry, Ultraviolet; D013379:Substrate Specificity
chemical_list: D018076:DNA, Complementary; D004271:DNA, Fungal; D005656:Fungal Proteins; D006023:Glycoproteins; D012331:RNA, Fungal; D042845:Laccase
keywords: 454 pyrosequencing; Dye decolorisation; Gene isolation; Inverse PCR; Laccase; Yellow laccase