Yellow laccase from the fungus Pleurotus ostreatus D1: Purification and characterization

Pozdnyakova, N. N.; Turkovskaya, O. V.; Yudina, E. N.; Rodakiewicz-Nowak, Ya.

doi:10.1134/s000368380601008x

Cited by 56 publications

(45 citation statements)

References 24 publications

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“…As a positive control for cellulase and xylanase production, a rich medium for basidiomycetes (RMB) supplemented with CM-cellulose (RMB-CMC) and xylan (RMB-Xyl) at a concentration of 2% (w/v) was used. The average composition as reported by Pozdnyakova et al (2006) was 10 g of glucose, 0.5 g of yeast extract, 10 g of peptone, 0.72 g of NH4NO3, 1.0 g of MgSO4, 0.5g of KCl, and 1 mL of trace element solution (1 g/L FeSO4, 2.8 g/L ZnSO4, and 3.3 g/L CaCl2) per 100 mL double-distilled water. Submerged fermentation of the residues was performed using 60-mM potassium phosphate buffer at pH 6.…”

Section: Enzyme Production In Submerged Mediummentioning

confidence: 99%

Selection and Characterization of a Native Pycnoporus sanguineus Strain as a Lignocellulolytic Extract Producer from Submerged Cultures of Various Agroindustrial Wastes

et al. 2015

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A native strain of Pycnoporus sanguineus (CS2) was selected because of its lignocellulolytic potential on a color-based agar plate screening system; pectinolytic and amylolytic capabilities were also assessed. The effects of different lignocellulosic substrates under submerged cultures of selected fungi on the enzymatic production of cellulases, xylanases, amylases, and laccases were evaluated. Seven different treatments, in which different combinations of wheat straw (WS), wheat bran (WB), orange peels (OP), grapefruit peels (GP), and apple peels (AP) were used, were established. Controls of 2% (w/v) carboxymethyl-cellulose (CMC) and xylan were used in a rich medium for basidiomycetes (RMB). The highest titers were achieved using OP-based cultures, with large titers of CMCases (33.5 U), avicelases (15.7 U), ß-D-glucosidase (72.9U), and xylanases (18.3 U). The best levels of amylase and laccase activity were obtained in the RMB plus CMC (RMB-CMC) (7.2 U) and in the medium OF/AP/GP (6.4 U), respectively. The relative molecular sizes of cellulase, xylanase, and amylase were 66.2, 56.5, and 90.8 kDa, respectively. Laccase and amylase had maximum activities at 60 °C whereas cellulase and hemicellulase had maximum activities at 70 °C. The optimal pH for cellulases, xylanases, and amylases was 5.0 in every case, and more than 95% activity was observed at pH 6. These results reveal some efficient operating parameters for the application of these enzymatic extracts as adjuvants to improve animal nutrition.

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Section: Enzyme Production In Submerged Mediummentioning

confidence: 99%

Selection and Characterization of a Native Pycnoporus sanguineus Strain as a Lignocellulolytic Extract Producer from Submerged Cultures of Various Agroindustrial Wastes

et al. 2015

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“…The determination of production curves for cellulases, xylanases, and ligninases was performed in a rich medium for basidiomycetes as reported by Pozdnyakova et al (2006). Its composition per liter was: 10 g glucose; 0.5 g yeast extract; 10 g peptone; 0.72 g NH4NO3; 1.0 g MgSO4; 0.5 g KCl; and 1 mL of trace solution (1 g/L FeSO4; 2.8 g/L ZnSO4; 3.3 g/L CaCl2).…”

Section: Identification Of Enzymatic Profiles In Liquid Mediamentioning

confidence: 99%

Native Macrofungi that Produce Lignin-Modifying Enzymes, Cellulases, and Xylanases with Potential Biotechnological Applications

et al. 2015

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With the aim of identifying and exploiting the mycological resources available in the Mexican Sierra Madre Oriental, the lignocellulolytic and pectinolytic potential of autochthonous fungi were evaluated in the present work. A solid media selection system was established in which 74 isolated strains were tested and compared to six international reference strains. The macrofungi Xylaria sp CS121, Inonotus sp CU7, Basidiomycete CH32, Basidiomycete CH23, Xylaria poitei, and Trametes maxima CU1 showed the highest cellulolytic and pectinolytic potential. The greatest lignolytic capability was exhibited by T. maxima CU1 and Pycnoporus sanguineus CS43. Under stirred submerged culture, T. maxima CU1 (cellulases, cellobiose dehydrogenase, manganese peroxidase (MnP), and laccase, with 200, 359, 51, and 267 U/L, respectively) and Xylaria sp CS121 (198 U/L of xylanases) were the highest enzymatic producers. Under stationary conditions, the best producers were Inonotus sp CU7 for cellulases, P. sanguineus CS43 for cellobiose dehydrogenase and laccase, and T. maxima CU1 for xylanases and MnP (242, 467, 35, 165, and 31 U/L, respectively). These results demonstrate the efficiency of enzymatic profiling as a tool for enzyme discovery with Mexican native fungi.

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“…The A280/A610 ratio represents the combined absorbances of tryptophan and aromatic residues at 280 nm, divided by the absorbance of T1 Cu at 610 nm. Generally, blue laccases exhibit an A280/A610 ratio of ~20, while that of the yellow laccases ranges from 90 to 150 (Leontievsky et al 1997a;Pozdnyakova et al 2006a). Thus, the PM1L-wt is a blue-laccase, while the OB-1 mutant can be considered as yellow laccase, based on its T1 Cu spectral features ( Table 1).…”

Section: The Yellow Ob-1 Mutantmentioning

confidence: 99%

“…Laccase-mediator systems have been exhaustively investigated, as they permit the oxidation of high redox potential compounds that cannot be oxidized by laccases alone (including lignin derivatives, recalcitrant dyes and PAHs) (Morozova et al 2007). Some fungal laccases produced in solid-phase cultures are yellow rather than the typical blue colour exhibited by the same laccases when produced in liquid cultures (Leontievsky et al 1997a(Leontievsky et al , 1997bPozdnyakova et al 2004Pozdnyakova et al , 2006a. In solid state fermentation of white-rot fungi using lignocellulosic materials (e.g., wheat straw containing lignin), a natural lignin-derived compound can modify the T1 Cu, switching from the oxidized resting state (Cu 2+ ) to the reduced state (Cu 1+ ), and thus quenching the Abs610T1Cu.…”

Section: Laccase Mediator Systemmentioning

confidence: 99%

“…In addition to blue laccases, several other laccases with distinct spectral characteristics have been described, such as yellow-brown laccases obtained from white-rot fungi under solid-state fermentation conditions. These laccases do not contain Abs610T1Cu, supposedly due to the presence of a putative natural mediator derived from lignin biodegradation located at the T1 Cu site (Leontievsky et al 1997a(Leontievsky et al , 1997bPozdnyakova et al 2004Pozdnyakova et al , 2006a). White laccases produced by different basidiomycete strains have also been described with a broad band at around 400 nm but no absorbance at 610 nm.…”

Section: Introductionmentioning

confidence: 99%

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Switching from blue to yellow: altering the spectral properties of a high redox potential laccase by directed evolution

Maté

García-Ruiz

Camarero

et al. 2012

Biocatalysis and Biotransformation

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During directed evolution to functionally express the high redox potential laccase from the PM1 basidiomycete in Saccharomyces cerevisiae (Mate et al. 2010), the characteristic maximum absorption at the T1 copper site (Abs610T1Cu) was quenched, switching the typical blue colour of the enzyme to yellow. To determine the molecular basis of this colour change, we characterized the original wild-type laccase and its evolved mutant. Peptide printing and Maldi-TOF analysis confirmed the absence of contaminating protein traces that could mask the Abs610T1Cu, while conservation of the redox potential at the T1 site was demonstrated by spectroelectrochemical redox titrations. Both wild-type and evolved laccases were capable of oxidizing a broad range of substrates (ABTS, guaiacol, DMP, synapic acid) and they displayed similar catalytic efficiencies. The laccase mutant could only oxidize high redox potential dyes (Poly R478, Reactive Black 5, Azure B) in the presence of exogenous mediators, indicating that the yellow enzyme behaves like a blue laccase. The main consequence of over-expressing the mutant laccase was the generation of a six-residue N-terminal acidic extension, which was associated with the failure of the STE13 protease in the Golgi compartment giving rise to alternative processing. Removal of the N-terminal tail had a negative effect on laccase stability, secretion and its kinetics, although the truncated mutant remained yellow. The results of CD spectra analysis suggested that polyproline helixes were formed during the directed evolution altering spectral properties. Moreover, introducing the A461T and S426N mutations in the T1 environment during the first cycles of laboratory evolution appeared to mediate the alterations to Abs610T1Cu by affecting its coordinating sphere. This laccase mutant is a valuable departure point for further protein engineering towards different fates.

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Yellow laccase from the fungus Pleurotus ostreatus D1: Purification and characterization

Cited by 56 publications

References 24 publications

Selection and Characterization of a Native Pycnoporus sanguineus Strain as a Lignocellulolytic Extract Producer from Submerged Cultures of Various Agroindustrial Wastes

Selection and Characterization of a Native Pycnoporus sanguineus Strain as a Lignocellulolytic Extract Producer from Submerged Cultures of Various Agroindustrial Wastes

Native Macrofungi that Produce Lignin-Modifying Enzymes, Cellulases, and Xylanases with Potential Biotechnological Applications

Switching from blue to yellow: altering the spectral properties of a high redox potential laccase by directed evolution

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