title: Characterization of cellobiose dehydrogenase and its FAD-domain from the ligninolytic basidiomycete Pycnoporus sanguineus.
abstract: Cellobiose dehydrogenase (CDH), an extracellular flavocytochrome produced by several wood-degrading fungi, was detected in the culture supernatant of the selective delignifier Pycnoporus sanguineus maintained on a cellulose-based liquid medium. Cellobiose dehydrogenase was purified as two active fractions: CDH1-FAD (flavin domain) (40.4 fold) with recovery of 10.9% and CDH1 (flavo-heme enzyme) (54.7 fold) with recovery of 9.8%. As determined by SDS-PAGE, the molecular mass of the purified enzyme was found to be 113.4kDa and its isoelectric point was 4.2, whereas these values for the FAD-domain were 82.7kDa and pI=6.7. The carbohydrate content of the purified enzymes was 9.2%. In this work, the cellobiose dehydrogenase gene cdh1 and its corresponding cDNA from fungus P. sanguineus were isolated, cloned, and characterized. The 2310bp full-length cDNA of cdh1 encoded a mature CDH protein containing 769 amino acids, which was preceded by a signal peptide of 19 amino acids. Moreover, both active fractions were characterized in terms of kinetics, temperature and pH optima, and antioxidant properties.
authors: Department of Biochemistry, Maria Curie-Skłodowska University, Akademicka 19 St., 20-033 Lublin, Poland.
authors: Justyna
authors: J
authors: Sulej
authors: Grzegorz
authors: G
authors: Janusz
authors: Monika
authors: M
authors: Osińska-Jaroszuk
authors: Paweł
authors: P
authors: Małek
authors: Andrzej
authors: A
authors: Mazur
authors: Iwona
authors: I
authors: Komaniecka
authors: Adam
authors: A
authors: Choma
authors: Jerzy
authors: J
authors: Rogalski
mesh_terms: D000975:Antioxidants / Q000737:chemistry / Q000378:metabolism; D002237:Carbohydrate Dehydrogenases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004271:DNA, Fungal / Q000235:genetics; D004795:Enzyme Stability; D005182:Flavin-Adenine Dinucleotide / Q000737:chemistry; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D005800:Genes, Fungal; D007526:Isoelectric Point; D007700:Kinetics; D008031:Lignin / Q000378:metabolism; D008970:Molecular Weight; D017434:Protein Structure, Tertiary; D055494:Pycnoporus / Q000201:enzymology* / Q000235:genetics; D013696:Temperature
chemical_list: D000975:Antioxidants; D004271:DNA, Fungal; D005656:Fungal Proteins; D005182:Flavin-Adenine Dinucleotide; D008031:Lignin; D002237:Carbohydrate Dehydrogenases; C019859:cellobiose-quinone oxidoreductase
keywords: Cellobiose dehydrogenase; Fungi; Gene; Pycnoporus