Characterization of Cellobiose Dehydrogenase from a Biotechnologically Important Cerrena unicolor Strain

Abstract

Cellobiose dehydrogenase (CDH), a secreted flavocytochrome produced by a number of wood-degrading fungi, was detected in the culture supernatant of a biotechnologically important strain of Cerrena unicolor grown in a modified cellulose-based liquid medium. The enzyme was purified as two active fractions: CuCDH-FAD (flavin domain) (1.51-fold) with recovery of 8.35 % and CuCDH (flavo-heme enzyme) (21.21-fold) with recovery of 73.41 %. As CDH from other wood-rotting fungi, the intact form of cellobiose dehydrogenase of C. unicolor is a monomeric protein containing one flavin and one heme b with molecular mass 97 kDa and pI = 4.55. The enzyme is glycosylated (8.2 %) mainly with mannose and glucosamine residues. Moreover, the cellobiose dehydrogenase gene cdh1 and its corresponding cDNA from the fungus C. unicolor were isolated, cloned, and characterized. The 2316-bp full-length cDNA of cdh1 encoded a mature CDH protein containing 771 amino acids preceded by a signal peptide consisting of 18 amino acids. Moreover, both active fractions were characterized in terms of kinetics, temperature and pH optima, and antioxidant properties.

The wood decay fungus Cerrena unicolor adjusts its metabolism to grow on various types of wood and light conditions

“…Moreover, strains of C . unicolor are also well described as potential bioproducers of industrially relevant enzymes such as extracellular laccase, manganese peroxidase, versatile peroxidase [10–13], cellobiose dehydrogenase [14], xylanase, and cellulase [15]. It has also been demonstrated recently that C .…”

Natural microbial polysaccharides as effective factors for modification of the catalytic properties of fungal cellobiose dehydrogenase

“…Janczarek ( 2011) and Osińska-Jaroszuk et al (Sigma Chemical Co., St. Louis, MO, USA) as electron acceptor according to the method proposed by Baminger et al (Baminger et al 2001) with modifications (Sulej et al 2015). The analysis was conducted in 48 well microplates using a Tecan Infinite M200 Pro (Tecan, Zürich, Switzerland) plate reader.…”

“…The antioxidant properties of CDH in the presence of PSs from the studied microorganisms was investigated using the DPPH assay according to an adapted spectrophotometry procedure described by Paduch et al (2008) with slight modifications. Lactose or cellobiose was added to the reaction mixture as substrates of the enzyme, as described previously (Sulej et al 2013(Sulej et al , 2015. Standards (Trolox and Vitamin C) that are well known for their strong antioxidant activity were used as a positive control.…”

Light-regulated synthesis of extra- and intracellular enzymes related to wood degradation by the white rot fungus Cerrena unicolor during solid-state fermentation on ash sawdust-based medium

“…Cerrena unicolor, a mossy maze polypore, is a globally distributed wood-degrading basidiomycete that causes extensive white rot (Enebak & Blanchette, 1989;Roody, 2003). C. unicolor strains are also well described as potential bioproducers of industrially relevant enzymes and other bioactive compounds of medical importance (Janusz et al, 2007;Hibi et al, 2012;Belova et al, 2014;Mizerska-Dudka et al, 2015;Sulej et al, 2015). A recent RNAseq-based transcriptomic analysis has provided new Vol.…”