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Microencapsulation is an alternative to increase the survival capacity of microorganisms, including Yarrowia lipolytica, a widely studied yeast that produces high-value metabolites, such as lipids, aromatic compounds, biomass, lipases, and organic acids. Thus, the present study sought to investigate the effectiveness of different wall materials and the influence of the addition of salts on the microencapsulation of Y. lipolytica, evaluating yield, relationship with cell stability, ability to survive during storage, and in vitro application of ruminant diets. The spray drying process was performed via atomization, testing 11 different compositions using maltodextrin (MD), modified starch (MS) and whey protein concentrate (WPC), Y. lipolytica (Y. lipo) cells, tripolyphosphate (TPP), and sodium erythorbate (SE). The data show a reduction in the water activity value in all treatments. The highest encapsulation yield was found in treatments using MD + TPP + Y. lipo (84.0%) and WPC + TPP + Y. lipo (81.6%). Microencapsulated particles showed a survival rate ranging from 71.61 to 99.83% after 24 h. The treatments WPC + Y. lipo, WPC + SE + Y. lipo, WPC + TPP + Y. lipo, and MD + SE + Y. lipo remained stable for up to 105 days under storage conditions. The treatment WPC + SE + Y. lipo (microencapsulated yeast) was applied in the diet of ruminants due to the greater stability of cell survival. The comparison between the WPC + SE + Y. lipo treatment, wall materials, and the non-microencapsulated yeast showed that the microencapsulated yeast obtained a higher soluble fraction, degradability potential, and release of nutrients.

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Oil emulsified in water is one of the most difficult mixtures to treat due to the good stability of emulsions, so there is a growing demand for more efficient methods for separating immiscible oil/water mixtures. In this context, the focus of this study was to obtain an adsorbent for the selective treatment of a simulated oily wastewater. To this aim, a modified hydrotalcite sample with hydrophobic and magnetic characteristics was prepared and characterized. Initially, the effect of sodium dodecyl sulfate (SDS) amount on the adsorbent characteristics was evaluated (266-800 mgSDS g-1LDH). The hydrophobic hydrotalcite (LDH-SDS) containing 533 mgSDS g-1LDH (LDH-SDS2) presented a higher interlayer space where the surfactant molecules were arranged perpendicular to the lamellae, allowing better access to the hydrotalcite pores and facilitating the selective adsorption of oil compounds. Moreover, the synergistic association of hydrophobic properties with super-wetting and effective adhesion oil to Fe3O4 favoured the selective adsorption of the simulated oily wastewater onto the hydrophobic and magnetic hydrotalcite (LDH-MSDS), facilitating the post-treatment separation. The kinetic analysis demonstrated that the adsorption equilibrium was attained in 120 min and the pseudo-second order model was the most suitable for predicting the removal of total organic carbon (TOC) from the simulated oily wastewater. The Langmuir model described very well the equilibrium experimental data, with a maximum adsorption capacity for TOC removal using LDH-MSDS of 659.9 mg g-1. Therefore, the modified hydrotalcite prepared in this study showed intrinsic characteristics that make it a promising adsorbent for the selective treatment of oily wastewaters.

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This work presents the immobilization in situ of commercial lipase from Candida antarctica B (CALB) by the sol-gel technique (xerogel) using silica from rice husk ash (RHA) as a source of silicon. It was used the Ionic Liquid (IL) 1-octyl-3-methylimidazolium bromide (C8MI.Br) as additive. The immobilized derivatives were characterized per SEM, XRD, and per method BET. The enzymatic activity of xerogels was evaluated with different tests, these being the reactional thermal analysis, immobilization yield, and operational and storage stability. The XDR showed that the obtained xerogels have halos in the region between 15 and 35° (2θ) what characterizes it as amorphous materials. The SEM analysis of xerogel shows irregular particles with dimensions less than 20 µm. The immobilized presented an esterification activity (EA) with 263.2 and 213.8 U/g, with and without IL, respectively, higher than the free enzyme (169.6 U/g). The immobilized, with and without IL, presented a significant improvement in the activity performance in relation to free enzyme for the three reactional temperatures (40, 60, and 80 °C) evaluated. The operational stability demonstrated that is possible to use xerogel without ionic liquid for 17 recycles and 21 recycles in IL presence. This methodology allows the preparation of new highly active and selective enzyme catalysts using the rice husk ash as a source of silicon, and the ionic liquid [C8MI]Br as additive. Furthermore, the new materials can provide greater viability in the processes, ensuring longer catalyst life.

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MCM-48 mesoporous support was synthesized with the ionic solid 1-tetradecyl-3-methylimidazolium chloride ([C14MI]Cl) as a structure-directing agent for in situ immobilization of Candida antarctica B (CALB). The MCM-48[C14MI]Cl support showed characteristics of mesoporous material of interest, with a pore size of 20.30 and 73.41 A for the support without and with the enzyme, respectively. The elongation of the carbonic chain of the ionic solid directly influenced the increase in the specific area and pore volume of the material. In addition, the decrease in the specific area and pore volume for support with the enzyme showed the effectiveness of immobilization in situ. It was possible to obtain the ideal levels for the best activities of esterification of the enzyme with optimization of a mathematical model. The optimized variables were 0.31 g of enzyme and 3.35% of ionic solid with a maximum esterification activity of 392.92 U/g and 688% of yield. The support showed residual activity above 50% when stored under refrigeration for 75 days. At 60 and 80 °C, the enzyme immobilized on the support retained more than 80 and 40% of its residual activity, respectively. In addition, the support presented the possibility of reuse for up to 10 cycles with residual activity of approximately 50%. The support synthesized in the present study presents a great industrial opportunity for the immobilization and use of the CALB enzyme.

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Sol-gel technique aiming enzymatic immobilization in situ with ionic liquids as additives is poorly studied. In this process, the addition of the enzyme is carried out in the synthesis of the support. The characteristics of ionic liquids, such as low vapor pressure, thermal stability, and non-flammability, make them strong candidates for use as immobilization additives. The objective of the present study was to immobilize the Candida antarctica B lipase by the sol-gel technique using ionic liquids as additives. The optimum points determined for ionic liquids 1-butyl-3-methylimidazolium chloride, 1-octyl-3-methylimidazolium bromide, and 1 hexadecyl-3-methylimimidazolium were 0.30, 0.27, and 0.22 g/mL of enzyme and 1.60, 1.52, and 1.52% of additive, respectively. The amount of enzyme and ionic liquids used in aerogel immobilization was the same as the optimized values in the xerogel immobilization process (for each ionic liquid). Ionic liquids proved to be good additives in the enzymatic immobilization process. Xerogel, regardless of the ionic liquid, presented a greater number of use cycles and better thermal stability compared to aerogel.

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MCM-41 and MCM-48 with niobium were successfully synthesized using 1-tetradecyl-3-methylimidazolium chloride ([C14MI]Cl) as a structure-directing agent. The best Si/Nb molar ratio was chosen (Si/Nb = 20) and the CALB enzyme was immobilized in situ in the synthesized Nb-MCM. SEM micrographs showed the formation of very regular spherical agglomerates with a diameter between 0.25 and 0.75 µm. The material presented a surface area of 954 and 704 m2/g and a pore volume of 0.321 and 0.286 cm3/g, for Nb-MCM-41 and Nb-MCM-48, respectively. Also, both materials showed a pore size of 2.261 nm. The number of recycles obtained for the CALB enzyme immobilized in Nb-MCM-41 and Nb-MCM-48 was 26 recycles with a residual activity of 49.62% and 16 recycles with a residual activity of 53.01%, respectively. For both materials, enzymatic activity remained stable for 5 months of storage at room temperature and refrigeration. The supports were able to catalyze the esterification reaction at 40, 60, and 80 °C, showing industrial application in reactions that require high temperatures. This methodology allows the preparation of new highly active and selective enzyme catalysts using niobium and [C14MI]Cl. Also, the new materials can provide greater viability in processes, ensuring a longer service life of catalysts. Graphical abstract.

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In this work was optimized the production of benzyl cinnamate by enzymatic catalysis using the immobilized lipase NS88011 and to evaluate its biological properties. The optimized condition for this system was 1:3 (acid:alcohol) molar ratio, 59 °C, biocatalyst concentration 4.4 mg.mL-1 for 32 h, with a yield of 97.6 %. The enzyme stability study showed that the enzyme remains active and yields above 60 % until the 13th cycle (416 h), presenting a promising half-life. In the determination of the antioxidant activity of the ester, an inhibitory concentration necessary to inhibit 50 % of the free radical 2,2-diphenyl-1-picryl-hydrazyl DPPH (IC50) of 149.8 mg.mL-1 was observed. For acute toxicity against bioindicator Artemia salina, lethal doses (LD50) of 0.07 and 436.7 µg.mL-1 were obtained for the ester and cinnamic acid, showing that benzyl cinnamate had higher toxicity, indicating potential cytotoxic activity against human tumors.

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This study aimed to evaluate the effects of adding probiotic culture (Bifidobacterium animalis subsp. Lactis Bb-12) and prebiotics (fructooligosaccharide - FOS) to yoghurt formulations stored at 4°C for 28 days, using an experimental design (independent variables: (0-3% of FOS and probiotic starter cultures 0-3%). The pH, acidity, fat, syneresis, protein, ºBrix, sugars, FOS and probiotic bacteria count were analyzed. The probiotic- and prebiotic-added yoghurt formulations showed lower acidity, syneresis and glucose than the control yoghurt and compared to formulations containing probiotic and prebiotic separately. The 3% probiotic and prebiotic formulation showed a lower loss of concentration of FOS, and after 28 days presented 1.5g of FOS per 100g (0.3% kestose, 0.7% nystose, 0.5% fructosyl-nystose). Furthermore, the addition of prebiotics exerted a protective effect on probiotic bacteria, enhancing their survival.

Este estudo teve como objetivo avaliar os efeitos da adição de cultura probiótica (Bifidobacterium animalis subsp. Lactis Bb-12) e prebióticos (fructooligosacarídeo - FOS) a formulações de iogurte armazenadas a 4°C por 28 dias, utilizando um planejamento experimental (variáveis independentes: (0-3% de FOS e cultura probiótica starter 0-3%). Foram analisados pH, acidez, gordura, sinérese, proteína, ºBrix, açúcares, FOS e contagem de bactérias probióticas. As formulações de iogurte adicionado de probiótico e prebiótico apresentaram menor acidez, sinérese e glicose quando comparados ao iogurte controle e também em comparação com as formulações contendo probiótico e prebiótico sozinhas. A formulação com 3% de probiótico e prebiótico apresentou menor perda de concentração de FOS e, após 28 dias, apresentou 1,5g de FOS por 100g (0,3% de kestose, 0,7% de nystose , 0,5% de fructosil-nistose). Além disso, a adição de prebióticos exerceu um efeito protetor sobre as bactérias probióticas e aumentou a sua sobrevivência.

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ABSTRACT: This study aimed to evaluate the effects of adding probiotic culture (Bifidobacterium animalis subsp. Lactis Bb-12) and prebiotics (fructooligosaccharide - FOS) to yoghurt formulations stored at 4°C for 28 days, using an experimental design (independent variables: (0-3% of FOS and probiotic starter cultures 0-3%). The pH, acidity, fat, syneresis, protein, ºBrix, sugars, FOS and probiotic bacteria count were analyzed. The probiotic- and prebiotic-added yoghurt formulations showed lower acidity, syneresis and glucose than the control yoghurt and compared to formulations containing probiotic and prebiotic separately. The 3% probiotic and prebiotic formulation showed a lower loss of concentration of FOS, and after 28 days presented 1.5g of FOS per 100g (0.3% kestose, 0.7% nystose, 0.5% fructosyl-nystose). Furthermore, the addition of prebiotics exerted a protective effect on probiotic bacteria, enhancing their survival.

RESUMO: Este estudo teve como objetivo avaliar os efeitos da adição de cultura probiótica (Bifidobacterium animalis subsp. Lactis Bb-12) e prebióticos (fructooligosacarídeo - FOS) a formulações de iogurte armazenadas a 4°C por 28 dias, utilizando um planejamento experimental (variáveis independentes: (0-3% de FOS e cultura probiótica starter 0-3%). Foram analisados pH, acidez, gordura, sinérese, proteína, ºBrix, açúcares, FOS e contagem de bactérias probióticas. As formulações de iogurte adicionado de probiótico e prebiótico apresentaram menor acidez, sinérese e glicose quando comparados ao iogurte controle e também em comparação com as formulações contendo probiótico e prebiótico sozinhas. A formulação com 3% de probiótico e prebiótico apresentou menor perda de concentração de FOS e, após 28 dias, apresentou 1,5g de FOS por 100g (0,3% de kestose, 0,7% de nystose , 0,5% de fructosil-nistose). Além disso, a adição de prebióticos exerceu um efeito protetor sobre as bactérias probióticas e aumentou a sua sobrevivência.

RESUMO

This study aimed to evaluate the effects of adding probiotic culture (Bifidobacterium animalis subsp. Lactis Bb-12) and prebiotics (fructooligosaccharide - FOS) to yoghurt formulations stored at 4°C for 28 days, using an experimental design (independent variables: (0-3% of FOS and probiotic starter cultures 0-3%). The pH, acidity, fat, syneresis, protein, ºBrix, sugars, FOS and probiotic bacteria count were analyzed. The probiotic- and prebiotic-added yoghurt formulations showed lower acidity, syneresis and glucose than the control yoghurt and compared to formulations containing probiotic and prebiotic separately. The 3% probiotic and prebiotic formulation showed a lower loss of concentration of FOS, and after 28 days presented 1.5g of FOS per 100g (0.3% kestose, 0.7% nystose, 0.5% fructosyl-nystose). Furthermore, the addition of prebiotics exerted a protective effect on probiotic bacteria, enhancing their survival.(AU)

Este estudo teve como objetivo avaliar os efeitos da adição de cultura probiótica (Bifidobacterium animalis subsp. Lactis Bb-12) e prebióticos (fructooligosacarídeo - FOS) a formulações de iogurte armazenadas a 4°C por 28 dias, utilizando um planejamento experimental (variáveis independentes: (0-3% de FOS e cultura probiótica starter 0-3%). Foram analisados pH, acidez, gordura, sinérese, proteína, ºBrix, açúcares, FOS e contagem de bactérias probióticas. As formulações de iogurte adicionado de probiótico e prebiótico apresentaram menor acidez, sinérese e glicose quando comparados ao iogurte controle e também em comparação com as formulações contendo probiótico e prebiótico sozinhas. A formulação com 3% de probiótico e prebiótico apresentou menor perda de concentração de FOS e, após 28 dias, apresentou 1,5g de FOS por 100g (0,3% de kestose, 0,7% de nystose , 0,5% de fructosil-nistose). Além disso, a adição de prebióticos exerceu um efeito protetor sobre as bactérias probióticas e aumentou a sua sobrevivência.(AU)

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