Proteolytic and nematicidal potential of the compost colonized by Hypsizygus marmoreus.

Spent mushroom compost (SMC) is a residue generated in edible mushrooms production, such as Hypsizygus marmoreus. Its genome was recently sequenced, demonstrating cuticle-degrading protease genes. The present work aims to investigate the proteases from H. marmoreus spent mushroom compost (SMC) by verifying its action on nematode larvae. The extraction of the crude extract directly with water from H. marmoreus SMC proved to be efficient for proteases obtainment, with proteolytic activity of 195.36 ±â€¯18.38 U g-1 of compound. Moreover, the zymogram and SDS-PAGE indicated the presence of two proteases with estimated molecular weights of 30.2 and 33.7 kDa. Due to the protease activity present in H. marmoreus SMC extract, there was a significant reduction in the number of Panagrellus redivivus and L3 in treated group compared to control group (p < 0.01), with 52% and 26% of reduction, respectively. A0A151VWY3 mature protein is composed of 296 amino acid residues, exhibiting molecular weight and pI of 29.5 kDa and 6.72. A0A151WD28 mature protein is composed of 343 amino acid residues, exhibiting molecular weight and pI of 34.4 kDa and 8.04. In the present work it was demonstrated that SMC from H. marmoreus has easily extracted protease content, presenting two proteases, possibly with cuticle-degrading activity, which had significant nematicidal effect on P. redivivus and bovine infective larvae.

In vitro and in silico characterization of a novel dextranase from Pochonia chlamydosporia.

The objective of this study was to purify, characterize, and phylogenetically and structurally analyze the dextranase produced by the fungus Pochonia chlamydosporia. Dextranase produced by the fungus P. chlamydosporia was purified to homogeneity in two steps, with a yield of 152%, purification factor of 6.84 and specific activity of 358.63 U/mg. Its molecular weight was estimated by SDS-PAGE at 64 kDa. The enzyme presented higher activity at 50 °C and pH 5.0, using 100 mM citrate-phosphate buffer, was inhibited by Ag1+, Hg2+, Cu2+, Mg2+, and presented KM of 23.60 µM. Mature dextranase is composed of 585 amino acids residues, with a predicted molecular weight of 64.38 kDa and pI 5.96. This dextranase showed a strong phylogenetic similarity when compared to Trichoderma harzianum dextranase. Its structure consists of two domains: the first composed by 15 ß strands, and the second composed by a right-handed parallel ß-helix.

Minimum cocktail of cellulolytic multi-enzyme complexes obtained from white rot fungi via solid-state fermentation.

The objective of this work was to optimize the total cellulase activity of the crude extract cocktails from five white rot fungi produced by solid-state fermentation, by means of the central composite design. The white rot fungi Pleurotus ostreatus PLO 06, Pleurotus eryngii PLE 04, Trametes versicolor TRAM 01, Pycnosporus sanguineus PYC 02 and Phanerochaete chrysosporium PC were tested. For optimization process aiming at the maximum value of total cellulase activity (FPAse), the multi-enzyme cellulase complexes (crude extracts) of each fungus were mixed simultaneously in different proportions. There was increase in FPAse activity for the cocktails formed by the extracts of the five fungi together, compared to the extracts of each fungus alone. The model presented the minimum cocktail of enzymes for maximum total cellulase activity, with 100.00 µL PYC; 100.00 µL PC; 100.00 µL PLO06; 100.00 µL PLE04 and 200 µL TRAM01. The maximum value found was of 304.86 U/L. The result of the cocktails was very relevant, showing that there is an enzymatic complementation in the extracts that should be further studied. Concentrated extract cocktails should also be evaluated for biomass saccharification.

Non-phytate phosphorus requirement for broilers from 8 to 21 days of age under heat stress conditions.

Two trials were conducted to determine the non-phytate phosphorus (nPP) requirement for broiler under heat stress. In both trials, birds were distributed in a completely randomized 4 × 2 factorial design with four nPP concentrations: 0.25, 0.35, 0.45, and 0.55%, and two Ca supply techniques: Ca fixed at 0.899% (CaF) or varying along with nPP aiming a 2:1 Ca to nPP ratio (CaV). Both trials had eight pens/treatment, with nine and five birds/pen for exp. 1 and exp. 2, respectively. nPP concentration had no effect on feed intake (FI), body weight gain (BWG), feed conversion ratio (FCR), nor fat deposition ratio (FDR). nPP levels showed a linear effect on protein deposition ratio (PDR) only for CaF diets. The nPP levels had a significant effect, regardless the technique adopted, on tibia phosphorus (TibP), which varied quadratically, on tibia calcium (TibCa) that increased quadratically and linearly, respectively, on CaF and CaV diets, and on tibia ash (TibAsh) that showed a quadratic effect for both. No effect was observed on Ca to P ratio in the tibia (TibCa:TibP). The nPP levels showed a linear increase effect over phosphorus intake (PI), phosphorus excreted (PE), and phosphorus retained (PR), and a linear decrease effect on phosphorus retention coefficient (PRC). Therefore, the nPP requirement for broilers from 8 to 21 days of age that provided better performance and bone variables were 0.250 and 0.484%, respectively, for CaF diets and 0.250 and 0.511%, respectively, for CaV diets.