期刊
PROCESS BIOCHEMISTRY
卷 47, 期 6, 页码 968-975出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.procbio.2012.03.006
关键词
Sclerotinia sclerotiorum; Yellow laccase; EPR spectroscopy; UV-vis spectroscopy
资金
- RSD (Romanian Ministry for Education and Research) [PCCE 140/2008]
- ACM [POSDRU/88/1.5/S/60185]
- ERASMUS
Reported here are the production, purification and characterization of a laccase from the phytophathogenic fungus Sclerotinia sclerotiorum. This laccase is identified by mass spectrometry with a sequence coverage of 74.9% (458/577 AA) revealing that the protein is identical or highly homologous to a predicted oxidoreductase from this species (A7EM18 in the Uniprot database); the closest homologous protein previously isolated from a fungus is the Melanocarpus albomyces, with only 35% identity. The UV-vis spectral features of this laccase classify it as a yellow one. The EPR spectrum nevertheless demonstrates resemblance to blue laccases - including the type 1 center not detectable in UV-vis spectra. The presence of type 3 coppers was proven by fluorescence spectrum and by 330 nm band in UV-vis. The purified laccase has an apparent molecular mass of 70 kDa and appears as a monomer. The values of K-M and k(cat) were determined for ABTS, 2,6-dimethoxyphenol, p-phenylenediamine and guaicol and are typical of a laccase. The optimal pH value is around 4 except for ABTS, for which activity is linearly increasing with acidity. The high laccase activity in liquid culture makes S. sclerotiorum a useful source of laccase for practical applications. (C) 2012 Elsevier Ltd. All rights reserved.
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