Partitioning of laccases derived from Cerrena unicolor and Pleurotus sapidus in polyethylene glycol – phosphate aqueous two–phase systems

Comparison of downstream processing methods in purification of highly active laccase

“…The repeated fed-batch mode applied for Cerrena unicolor cultivation proved to be very efficient towards the production of highly active laccase. The maximal activity of 6400 U/L was about four times higher than previously recorded for the conventional batch bioreactor cultivation [16] and for the shake flaks cultures of this fungus [25]. The achieved activity is also satisfactory in comparison to the values reported in the literature obtained for other fungal strains: 30.2 U/L for Phanerochaete chrysosporium during batch fermentation [31], 1403 U/L for Pleurotus ostreatus in packed-bed bioreactor [32] or 2123 U/L for Trametes versicolor in fluidized-bed bioreactor [33].…”

“…Aqueous two-phase extraction approach was used under the conditions described in previous study [16]. The obtained results concerning the phase volume, laccase activity and protein content, as well as calculated activity yields and purification folds are presented in Table 1.…”

“…The vessels consisted of mixing flasks combined with burettes of approximately 20 mL volume. Mixture components, in the form of stock solutions and culture liquid, were prepared according to procedure described by Blatkiewicz et al [16] and the following proportion was applied: 4.8 g of PEG 6000, 3.4 g KH 2 PO 4 and 6.8 g of culture liquid. The mixing was carried out in a thermostated incubator equipped with a magnetic stirrer at the temperature set to 25 °C.…”

“…Prinz et al [15] investigated Pleurotus sapidus laccase partitioning in PEG-phosphate system. Blatkiewicz et al [16] demonstrated that both PEG 400 and PEG 6000 systems allowed for a very strong concentration of Cerrena unicolor laccase, with yields exceeding 0.85 and 0.9, respectively. Interestingly, the enzyme concentrated in different phases, with PEG 400 in a polymer and in PEG 6000 in a salt phase.…”

Genomic Studies of White-Rot Fungus Cerrena unicolor SP02 Provide Insights into Food Safety Value-Added Utilization of Non-Food Lignocellulosic Biomass

“…White-rot fungi secrete a variety of intracellular and extracellular enzymes that degrade and utilize lignocellulose and naturally mineralize the lignin [3]. C. unicolor can produce high amounts of the enzyme without inductors [4], known as the potential bio-producer of industrially relevant enzymes such as laccase [5], manganese peroxidase [6], versatile peroxidase [7], cellobiose dehydrogenase [8], xylanase, and cellulase [9]. C. unicolor has a unique capacity to degrade recalcitrant resistant lignin polymers as well as a wide spectrum of aromatic pollutants that provide potential value in biomass utilization and organo-pollutant degradation [10,11].…”