Enhanced removal of phenol from biorefinery wastewater treatment using enzymatic and Fenton process.

The valorisation of biomass has been commonly carried out in biorefineries. The environmental concerns about these processes have not been intensely considered, demanding further investigations. Particularly, phenols are founded in high concentrations in biorefinery wastewater and are considered compounds of major concern. In this study, we evaluated the bioconversion of phenols by enzymatic treatment using the enzyme Horseradish peroxidase (HRP) and the Fenton process. The results showed an enzymatic phenol conversion of 97.5% at pH 7.0, enzyme activity of 0.8 U/mL and hydrogen peroxide concentration of 1.61 g/L. So as to enhance the treatment, we evaluate the Fenton reaction as a complementary process for further remaining phenol conversion. The best conditions for Fenton process were achieved using a hydrogen peroxide concentration and [H2O2]:[Fe] ratio of 3.90 g/L and 74, respectively, and the obtained phenol concentration in the treated wastewater was 0.11 mg/L. Chromatography analysis showed that 2-methoxyphenol was the majority compound in the original wastewater, which was subsequently precipitated by the enzymatic treatment. Furthermore, many physicochemical parameters were modified due to the treatment, such as biochemical oxygen demand, chemical oxygen demand and total organic carbon, with removal efficiencies of around 97, 49 and 46%, respectively. HRP combined with Fenton can be considered as an alternative methodology for the biorefinery wastewater treatment, especially regarding the phenols conversion.

Application of glycerol as carbon source for continuous drinking water denitrification using microorganism from natural biomass.

The contamination of water resources by nitrate is a global problem. Indeed, traditional treatment technologies are not able to remove this ion from water. Alternatively, biological denitrification is a useful technique for natural water nitrate removal. This study aimed to evaluate the use of glycerol as a carbon source for drinking water nitrate removal via denitrification in a reactor using microorganisms from natural biomass. The experiment was carried out in a continuous fixed bed reactor using immobilised microorganisms from the vegetal Phyllostachys aurea. The tests were started in batch mode to provide cells growth and further immobilisation on the support. Then, the treatment experiments were accomplished in an up-flow continuous reactor. Ethanol was used as the primary carbon source, and it was gradually replaced by glycerol. The C:N (carbon to nitrogen) ratio and the hydraulic residence time (HRT) were evaluated. It was possible to remove 98.14% of nitrate using a C:N ratio and HRT of 3:1 and 1.51 days, respectively. The results have demonstrated that glycerol is a potential carbon source for denitrification in a continuous reactor using immobilised cells from natural biomass.

Using natural biomass microorganisms for drinking water denitrification.

Among the methods that are studied to eliminate nitrate from drinking water, biological denitrification is an attractive strategy. Although several studies report the use of denitrifying bacteria for nitrate removal, they usually involve the use of sewage sludge as biomass to obtain the microbiota. In the present study, denitrifying bacteria was isolated from bamboo, and variable parameters were controlled focusing on optimal bacterial performance followed by physicochemical analysis of water adequacy. In this way, bamboo was used as a source of denitrifying microorganisms, using either Immobilized Microorganisms (IM) or Suspended Microorganisms (SM) for nitrate removal. Denitrification parameters optimization was carried out by analysis of denitrification at different pH values, temperature, nitrate concentrations, carbon sources as well as different C/N ratios. In addition, operational stability and denitrification kinetics were evaluated. Microorganisms present in the biomass responsible for denitrification were identified as Proteus mirabilis. The denitrified water was submitted to physicochemical treatment such as coagulation and flocculation to adjust to the parameters of color and turbidity to drinking water standards. Denitrification using IM occurred with 73% efficiency in the absence of an external carbon source. The use of SM provided superior denitrification efficiency using ethanol (96.46%), glucose (98.58%) or glycerol (98.5%) as carbon source. The evaluation of the operational stability allowed 12 cycles of biomass reuse using the IM and 9 cycles using the SM. After physical-chemical treatment, only SM denitrified water remained within drinking water standards parameters of color and turbidity.

Immobilization of laccase from Aspergillus oryzae on graphene nanosheets.

Laccase enzymes of Aspergillus oryzae were immobilized on graphene nanosheets by physical adsorption and covalent bonding. Morphological features of the graphene sheets were characterized via microscopy techniques. The immobilization by adsorption was carried out through contact between graphene and solution of laccase enzyme dissolved in deionized water. The adsorption process followed a Freundlich model, showing no tendency to saturation within the range of values used. The process of immobilization by covalent bonding was carried out by nitration of graphene, followed by reduction of sodium borohydride and crosslinking with glutaraldehyde. The process of immobilization by both techniques increased the pH range of activity of the laccase enzyme compared to the free enzyme and increased its operating temperature. On operational stability, the enzyme quickly loses its activity after the second reaction cycle when immobilized via physical adsorption, while the technique by covalent bonding retained around 80% activity after six cycles.

Substrate specificity and enzyme recycling using chitosan immobilized laccase.

The immobilization of laccase (Aspergillus sp.) on chitosan by cross-linking and its application in bioconversion of phenolic compounds in batch reactors were studied. Investigation was performed using laccase immobilized via chemical cross-linking due to the higher enzymatic operational stability of this method as compared to immobilization via physical adsorption. To assess the influence of different substrate functional groups on the enzyme's catalytic efficiency, substrate specificity was investigated using chitosan-immobilized laccase and eighteen different phenol derivatives. It was observed that 4-nitrophenol was not oxidized, while 2,5-xylenol, 2,6-xylenol, 2,3,5-trimethylphenol, syringaldazine, 2,6-dimetoxyphenol and ethylphenol showed reaction yields up 90% at 40 °C. The kinetic of process, enzyme recyclability and operational stability were studied. In batch reactors, it was not possible to reuse the enzyme when it was applied to syringaldazne bioconversion. However, when the enzyme was applied to bioconversion of 2,6-DMP, the activity was stable for eight reaction batches.