Then, 20 μL of this suspension was injected into the HPLC (Shimad

Then, 20 μL of this suspension was injected into the HPLC (Shimadzu, C18 reverse

phase and UV detection at 280 nm) and eluted with a gradient of acetonitrile and 0.175 (g/100 mL) orthophosphoric acid ( Makkar et al., 1997). We use phorbol-12-myristate 13-acetate (Sigma) as standard. The soluble protein and reducing sugar contents were determined according to the colorimetric methods described by Bradford (1976) and Miller (1959). For this analysis, 10 g of the crushed mushrooms was placed in Erlenmeyer flasks (125 mL) containing 25 mL sodium citrate buffer AG-014699 clinical trial (0.05 mol/L and pH 4.8). These flasks were kept in a shaker for 30 min at 150 rpm, and the extracts were filtered using Millipore membranes (Cavallazzi et al., 2004). This experiment was conducted using a completely randomized design with 5 replicates. The data were subjected to analysis

of variance, and the averages were compared by Tukey’s test (p < 0.05) using Saeg software (version 9.1, Federal University of Viçosa). The tannin concentrations Selumetinib datasheet observed in the jatropha seed cake (Fig. 1) were similar to those found in the fruit peel of J. curcas ( Makkar, Aderibigbe, & Becker, 1998). The greatest concentration of this compound was observed in the substrate with coffee husk and eucalypt bark ( Fig. 1). This may have been due to the presence of tannins in the eucalypt bark ( Vázquez, González-Alvarez, Santos, Freire, & Antorrena, 2009) and in the coffee husk ( Barcelos, Paiva, Pérez, Santos, & Cardoso, 2001). The thermal treatment Methamphetamine of the substrates decreased the tannin concentration by 46%

(Fig. 1). This result was similar to that observed in vegetables after cooking or autoclaving at 121 °C and 128 °C for different periods of time (Rehman & Shah, 2005). Regardless of the substrate, tannin degradation by P. ostreatus Plo 6 increased as a function of the incubation time, and the highest rate was observed between 15 and 30 d in the substrate with coffee husk and eucalypt bark ( Fig. 1). The high degradation rate of this compound was also observed in Pleurotus sp. cultivated in coffee husk for 60 d ( Fan et al., 2006) and this tannin degradation may be related to tannase activity (tannin acyl hydrolase, EC 3.1.1.20). The activity of this enzyme in the polyphenols degradation has been reported in Aspergillus and Penicillium ( Batra & Saxena, 2005). Thus, P. ostreatus can degrade the tannins in J. curcas seed cake and the other tested substrates. The acid phytic content in the jatropha seed cake (Fig. 2A) was lower than the percentage this acid found in the seed of J. curcas by Makkar et al. (1998). This result shows that a percentage this acid was in the oil. Although phytic acid is considered to be heat-stable (Deshpande & Damodaran, 1990), the sterilization of the substrates decreased in 20% the content of phytic acid (Fig. 2A).

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