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This study aimed to produce and characterise microparticles produced from barley residue proteins (BRP) enriched with ß-carotene. The microparticles were obtained by freeze-drying five emulsion formulations with 0.5% w/w whey protein concentrate and different concentrations of maltodextrin and BRP (0, 1.5, 3.0, 4.5 and 6.0% w/w), with the dispersed phase consisting of corn oil enriched with ß-carotene. The mixtures were mechanically mixed and sonicated, the formed emulsions were freeze-drying. The microparticles obtained were tested for encapsulation efficiency, humidity, hygroscopicity, apparent density, scanning electron microscopy (SEM), accelerated stability and bioaccessibility. Microparticles produced with the emulsion containing 6% w/w BRP had lower moisture content (3.47 ± 0.05%), higher encapsulation efficiency (69.11 ± 3.36%), bioaccessibility value of 84.1% and greater ß-carotene protection against thermal degradation. SEM analysis showed that microparticles had sizes ranging from 74.4 to 244.8 µm. These results show that BRP are viable for the microencapsulation of bioactive compounds by freeze-drying.

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Enzyme immobilization has been reported as a promising approach to improving parameters such as thermal stability, pH and reusability. In this study, a polyacrylamide cryogel functionalized with L-phenylalanine was prepared to be used in the adsorption of ß-glucosidase from Thermoascus aurantiacus, aiming at its separation and also its immobilization on the cryogel matrix. The enzyme was produced by solid state fermentation. First, the adsorption was studied as a function of the pH and the resulting yield (Y, %) and purification factor (PF, dimensionless) were determined (1.57-5.13 and 64.19-91.20, respectively). The PF and yield from eluate samples obtained at pH 3.0 were the highest (5.13 and 91.20, respectively). Then, ß-glucosidase was immobilized on the hydrophobic cryogel and the recovery activities (%) were determined as a function of temperature and in the presence of different saline solutions. The values ranged from 14.45 to 45.97. As expected, salt type and ionic strength affected the activity remained in the immobilized ß-glucosidase. The average bioreactor activity was 39.9 U/g of dry cryogel and its operational stability was measured, with no decrease in activity being observed during seven cycles. Kinetic parameters of free and immobilized enzyme were determined according to different models.

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This study proposed the development of a monolithic supermacroporous affinity column for direct capture of lactoperoxidase, a glycoprotein present in milk, whey, and colostrum, with several applications due to its wide antimicrobial activity. A poly(acrylamide)-based cryogel was produced by radical co-polymerization of monomers in frozen aqueous solution and activated with p-aminobenzenesulfonamide as a ligand for specific interaction with the lactoperoxidase. The axial liquid dispersion coefficients at different liquid flow rates were determined by measuring residence time distributions using the tracer pulse-response method. The axial dispersion coefficient was low and the height equivalent to theoretical plate was not dependent on the flow velocity. The adsorptive capacity of affinity cryogel was studied as a function of flow velocity and the best condition was 0.9 cm/min. The response surface methodology was applied to optimize the capture of the enzyme, as a function of pH and salt concentration. Higher purification factor value was found at a salt concentration of 80 mmol/L and pH of 8.0 (p < 0.05). There was no influence of the variables under study on the yield (p > 0.05). The results indicated that affinity cryogel is a promising chromatography support for the use in high-throughput one-step purification of lactoperoxidase from whey.

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This study focused on the extraction, purification, and physicochemical characterization of γ-conglutin, a protein present in lupin seeds with properties of reducing blood glucose levels. Total protein was extracted with an alkaline-saline solvent, followed by isoelectric precipitation. Chromatographic purification of the precipitated fraction was performed using a cation exchange supermacroporous cryogel column. Electrophoresis of the eluted fraction from chromatography presented a single band of ∼48 kDa under non-reducing conditions (two bands of ∼30 and ∼17 kDa, under reducing conditions) confirming the success of the purification protocol. Liquid chromatography-tandem mass spectrometry analysis confirmed the identity of the protein as γ-conglutin. The purified γ-conglutin had an isoelectric point of 7.51, ß-sheets prevailing as a secondary structure, and denaturation temperature close to 68°C. The outcome of this work showed that γ-conglutin was obtained with a high degree of purity. The proposed purification protocol is simple and can be easily scaled up.

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Lectins are glycoproteins that bind to carbohydrates or glycoconjugates by specific interactions. The specificity of lectins to various carbohydrates is a determinant factor in the choice of ligand for the chromatographic matrix when using chromatography as a lectin purification technique. In this work, the immobilization of three different aminated carbohydrates on the surface of macroporous polymeric cryogels was evaluated. Carbohydrates were immobilized on cryogel surfaces via the glutaraldehyde method to create spacer arms, reducing steric hindrance. The immobilized N-acetyl-d-glucosamine and N-acetyl-d-mannosamine concentrations contained approximately 130mg of carbohydrate/g dehydrated cryogel, while the N-acetyl-d-galactosamine contained 105mg of carbohydrate/g dehydrated cryogel. Scanning electron microscopy showed that the physical structure and porosity of the chromatographic columns were not affected by the immobilization process, maintaining an elevated hydration capacity and the macroporous structure of the cryogels. Adsorption of concanavalin A on cryogels functionalized with N-acetyl-d-glucosamine (cryo-d-GlcNAc) was tested, as well as its reuse capability. After 5 cycles of use, cryo-d-GlcNAc was shown to be stable, with an adsorptive capacity of around 50mg/g. Carbohydrate immobilization in polyacrylamide cryogels was satisfactory, with promise for applications in lectin purification processes.

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In this study, a supermacroporous polyacrylamide cryogel was produced by cryo-polymerization and activated with Tris(hydroxymethyl)aminomethane (Tris-cryogel) to be applied as an affinity ligand for a one step purification of lysozyme (LYZ), directly from chicken egg white (EW). The Tris-cryogel presented interconnected pores with size varying in the range of 20-80µm and swelling capacity of 19.6±0.9g/g. The axial dispersion of the Tris-cryogel was analyzed at different flow velocities and mobile phase viscosities. It was verified that higher viscosity resulted in a higher degree of dispersion, causing the HETP values to increase from 0.04cm to 0.8cm. Adsorption isotherms were measured at 15°C and 35°C at pH 7.5. A Langmuir model was fitted to the equilibrium data, with a maximum adsorptive capacity of 285mg/g at 15°C and 363mg/g at 35°C. Thermodynamic analysis based on the Van't Hoff relationship showed that the process was spontaneous and enthalpically driven. Lysozyme was purified directly from egg white in a one step purification process at different pH values (7.5, 8.5 and 9.5). Independent of the pH, the specificity of Tris-cryogel for lysozyme adsorption was confirmed. At pH 7.5, yield and purification fold were higher (30% and 45). In addition, the effect of the dilution rate on egg white and flow velocity were also analyzed and it was shown that flow velocity did not affected purification and column efficiency, and that diluting the egg white increased yield to 70% with a purification fold of 23. Results show Tris-cryogel is a promising matrix for use in high throughput purification of lysozyme from egg white.

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O objetivo deste trabalho foi avaliar a qualidade físico-química e microbiológica de ricotas consumidasna região sudoeste da Bahia e verificar informações nutricionais contidas nos rótulos dos produtos,de acordo com os padrões de rotulagem exigidos pela legislação e sua conformidade com a composiçãonutricional real obtida por meio de análises laboratoriais. Foram adquiridas 18 amostras de 3 marcas de ricotas comercializadas na região sudoeste da Bahia. Os resultados obtidos foram submetidos à Análisede Variância (ANOVA) e as médias foram comparadas entre si por meio do teste de Tukey, ao nível de 5% de probabilidade. Os resultados da ANOVA indicaram que as marcas de ricotas avaliadas apresentaram diferenças significativas entre si (pO,05). Todas as marcas apresentaram não conformidade entre os valores nutricionais declarados e osvalores reais determinados. Os resultados das análises microbiológicas indicaram a presença de coliformes termotolerantes e totais em todas as marcas analisadas, porém, em valores abaixo do limite máximo estabelecido na legislação. Todas as marcas apresentaram contagem de Staphylococcus aureus acima do limite máximo estabelecido pela legislação, enquanto que, a presença de bolores e leveduras não foi detectada em nenhuma das amostras. Tais resultados indicam a necessidade de se estabelecer padrões de qualidade físico-química e microbiológica específicos para a ricota. (AU)

The aim of this work was to evaluate the physico-chemical and microbiological quality of ricottaconsumed in the southwestern region of Bahia, and also check whether the nutritional information on thelabels of the products were in accordance with the standards for labeling required by law and their conformity with the real nutritional composition obtained through laboratory tests. Has been acquired 18samples of 3 ricotta brands sold in the southwestern region of Bahia. The results were subjected to analysis of variance (ANOVA) and means were compared using the Tukey test at 5% probability. The results of the ANOVA showed a significant difference between brands of ricotta evaluated (p <0.05) for the pH values, ash, salt and protein. Ali the samples had not in conformity with the declared nutrient values and real values determined. The results of microbiological analysis indicated the presence of fecal coliform and total in all brands tested, but in amounts below the threshold set in legislation. Ali samples showed Staphylococcus aureus above the limit established by the legislation, while the presence of yeastsand molds was not detected in any sample. These results indicate the need to establish specific qualitystandards to the ricotta. (AU)

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