No abstract

are multicopper oxidases of great importance in the industry due to their non-specificity and high oxidative potential. Laccases are useful to bleach synthetic dyes, oxidize phenolic compounds and degrade pesticides, among others. Hence, the objective of this work was to optimize low cost culture media for recombinant (rPOXA 1B) laccase production from Pleurotus ostreatus in Pichia pastoris. To this end, low cost nitrogen sources were studied, such as malt extract, isolated soy protein and milk serum. Following, two central composite designs (CCD) were performed. In CCD-1 different concentrations of glucose USP (0-13.35 gL -1 ), protein isolated soy protein (5-25 gL -1 ), malt extract (3.5-17.5 gL -1 ) and (NH 4 ) 2 SO 4 (1.3-6.5 gL -1 ) were evaluated. In CCD-2 only different concentrations of glucose USP (7.9-22 gL -1 ) and isolated soy protein (15.9-44.9 gL -1 ) were evaluated. CCD-2 results led to a One Factor Experimental design (OFED) to evaluate higher isolated soy protein (20-80 gL -1 ) concentrations. In all designs, (CCD-1, CCD-2 and OFED) CuSO 4 (0.16 gL -1 ) and chloramphenicol (0.1 gL -1 ) concentrations remained unchanged. For the OFED after sequential statistical optimization, an enzyme activity of 12,877.3 AE 481.2 UL À1 at 168 h was observed. rPOXA 1B activity increased 30.54 % in comparison with CCD-2 results. Final composition of optimized media was: 20 gL -1 glucose USP, 50 gL -1 isolated soy protein 90 % (w/w), 11.74 gL -1 malt extract, and 4.91 gL -1 (NH 4 ) 2 SO 4 . With this culture media, it was possible to reduce culture media costs by 89.84 % in comparison with improved culture media previously described by our group.

The purpose of this study was to demonstrate that methanol addition after glucose depletion has a positive effect on improving rPOXA 1B production under the control of pGap in P. pastoris. Four different culture media (A, B, C and D) were used to culture P. pastoris X33/pGapZαA-LaccPost-Stop (clone 1), containing a previously optimized POXA 1B synthetic gene coding for P. ostreatus laccase, which after glucose depletion was supplemented or not with methanol. Enzyme activity in culture media without methanol (A, B, C and D) was influenced by media components, presenting activity of 1254.30 ± 182.44, 1373.70 ± 182.44, 1343.50 ± 40.30 and 8771.61 ± 218.79 U L−1, respectively. In contrast, the same culture media (A, B, C and D) with methanol addition 24 h after glucose depletion attained activity of 4280.43 ± 148.82, 3339.02 ± 64.36, 3569.39 ± 68.38 and 14,868.06 ± 461.58 U L−1 at 192 h, respectively, representing an increase of approximately 3.9-, 2.4-, 3.3- and 1.6-fold compared with culture media without methanol. Methanol supplementation had a greater impact on volumetric enzyme activity in comparison with biomass production. We demonstrated what was theoretically and biochemically expected: recombinant protein production under pGap control by methanol supplementation after glucose depletion was successful, as a feasible laboratory production strategy of sequential carbon source addition, breaking the habit of utilizing pGap with glucose.

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The modified tannin by Mannich reaction was investigated for wastewater treatment. The removal of heavy metals, such as copper, chromium and mercury, in industrial wastewater was evaluated through the coagulation–flocculation technique, using modified Acacia tannin (MAT) as a coagulant agent. The successful tannin modification was evaluated by infrared spectopometry (FTIR), nuclear magnetic resonance (NMR); monitoring the removal of heavy metals was performed by atomic absorption (AA) and a direct mercury analyzer (DMA). Additionally, the parameters of water quality, total suspended solids (TSS), turbidity and chemical oxygen demand (COD) were assessed. Different doses of MAT were evaluated (375 ppm, 750 ppm, 1250 ppm and 1625 ppm) and three different levels of pH (4, 7 and 10). The highest percentages of removal obtained were copper 60%, chromium 87%, mercury 50%–80%, COD 88%, TSS 86% and turbidity 94%, which were achieved with the dose of 375 ppm of MAT at pH 10. The coagulation–flocculation process with the modified Acacia tannin is efficient for the removal of conventional parameters and for a significant removal of the metals studied.

The catalytic oxidation of aqueous crystal violet (CV) solutions was investigated using Ni and Fe catalysts supported over Mg–Al oxides synthesized by the autocombustion method. The influence of temperature, loading, and selectivity were studied in the catalytic wet air oxidation (CWAO) of CV. The kind of metal had an important contribution in the redox process as significant differences were observed between Fe, Ni, and their mixtures. The catalysts with only Fe as active phase were more efficient for the oxidation of CV under normal conditions (T = 25 °C and atmospheric pressure) compared to those containing Ni, revealing the influence of the transition metal on catalytic properties. It was found that iron-containing materials displayed enhanced textural properties. The synthesis of Fe/MgAl catalysts by the autocombustion method led to solids with excellent catalytic behavior, 100% CV degradation in eight hours of reaction, 68% selectivity to CO2, and significant reduction of chemical oxygen demand (COD).

Low-density polyethylene (LDPE) sheets (3.0 ± 0.1 cm) received sequential treatment, first by the action of direct-current low-pressure plasma (DC-LPP) with a 100% oxygen partial pressure, 3.0 × 10−2 mbar pressure, 600 V DC tension, 5.6 cm distance, 6-min treatment. Then, sheets were submitted to TiO2 photocatalysis at UV radiation at 254 nm (TiO2/UV) with a pH value of 4.5 ± 0.2 and a TiO2 concentration of 1 gL−1. We achieved a complementary effect on the transformation of LDPE films. With the first treatment, ablation was generated, which increased hydrophilicity. With the second treatment, the cavities appeared. The changes in the LDPE sheets’ hydrophobicity were measured using the static contact angle (SCA) technique. The photocatalytic degradation curve at 400 h revealed that the DC-LPP photocatalysis sequential process decreased SCA by 82°. This was achieved by the incorporation of polar groups, which increased hydrophilicity, roughness, and rigidity by 12 and 38%, respectively. These sequential processes could be employed for LDPE and other material biodegradation pretreatment.