Purification, kinetics and spectral characterisation of a new versatile peroxidase from a Bjerkandera sp. isolate

Abstract

From the extracellular fluid of a novel strain of Bjerkandera sp., it was isolated, purified and identified the main enzyme responsible for Remazol Brilliant Blue R dye decolourisation. Such an enzyme is able to oxidise manganese, as well as veratryl alcohol and 2,6dimethoxyphenol in manganese-independent reactions; hence, it can be included in the new group of versatile peroxidases. The molecular mass of said enzyme is ca. 45 kDa, and the N-terminal amino acid sequence obtained by Edman degradation is VAXPDGVNTA. The enzyme substrate range for oxidation of several phenolic and non-phenolic aromatic compounds was determined and the corresponding Michaelis-Menten kinetic constants calculated. Furthermore, spectrophotometric assays showing the Soret band and allowing observation of band shifts of the enzyme led to the conclusion that Bjerkandera strains may also synthesise at least two different versatile peroxidases, as happens with Pleurotus eryngii.

Recent developments in biodegradation of industrial pollutants by white rot fungi and their enzyme system

“…The deduced amino acid sequence of MnP2 isolated from P. ostreatus and VP (VAXPDGVNTA) from a novel strain of Bjerkandera sp. (MW 45 kDa) is more than 95% identical to that of a typical VP (VP-PS1) from P. eryngii (Moreira et al 2006). A homologous enzyme, MnP-GY, from recombinant strains P. ostreatus ATCC66376 (Kamitsuji et al 2004) and MnP2 from P. ostreatus TM2-10 (Tsukihara et al 2006) can directly oxidize veratryl alcohol and highmolecular weight compounds, such as Poly R-478 and RNaseA, although much less efficiently than it oxidized Mn 2+ in the absence of a mediator suggesting that the enzyme is a VP having properties of both LiP and MnP.…”

Reaction Kinetics of Versatile Peroxidase for the Degradation of Lignin Compounds

“…The RZ value for the crude VP was 0.3. At least a factor of ten lower, than RZ values for other purified VPs, ranging from 3.2 (Moreira et al, 2006) and 3.5 up to 4.0 (Garcia-Ruiz et al, 2012). However, the RZ value is no indicator for enzymatic activity necessarily (Dunford, 1991;SAC, 2012).…”

“…Whereas the reaction with guaiacol (VII) could be monitored, in principle, at 456 nm (Moreira et al, 2006) or at 470 nm as well, due to polymerization (tetraguaiacol (Chance and Maehly, 1955)) of its radical products, apparently phenoxy radicals (Lundell et al, 1993b), the reduction of E II to E 0 by component III results finally in veratraldehyde formation (Schoemaker, 1990;Schoemaker et al, 1994b). The latter may only an option under low oxygen conditions (Schoemaker et al, 1994b), since the existence of III is supposed to be shortlived by reacting with O 2 immediately (see discussion above).…”

Decolorization of Remazol Brilliant Blue (RBBR) and Poly R-478 dyes by Bjerkandera adusta CCBAS 930

“…These enzymes catalyze the oxidative depolymerization of lignin in ligninocellulose wood complexes [6], and also other aromatic compounds possessing a lignin-like structure such as synthetic dyes [7,8]. Among white-rot basidiomycetes growing on wood, the ability to biodegrade synthetic dyes was reported for Phanerochaete chrysosporium, Trametes (Coriolus) versicolor, Pleurotus ostreatus, P. eryngii, Phlebia radiata, and Bjerkandera adusta [9][10][11][12][13][14]. P. chrysosporium demonstrated the greatest effectiveness at decolorization and biodegradation of anthraquinonic, azo and triphenylmethans dyes [9,11,12].…”

“…P. chrysosporium demonstrated the greatest effectiveness at decolorization and biodegradation of anthraquinonic, azo and triphenylmethans dyes [9,11,12]. Decolorization and biodegradation abilities of different Bjerkandera spp., including B. adusta strains, were widely studied, and basically include azo, anthraquinonic, triphenylmethans and heterocyclic dyes [2,5,[13][14][15][16][17][18][19][20]. A new strain Bjerkandera adusta CCBAS 930 isolated from soil was described [21].…”