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Ficin fully immobilized on Asp-agarose beads at pH 7 but not on an aminated support. This made enzyme adsorption plus glutaraldehyde modification non-viable for this enzyme. Modifying glyoxyl-agarose beads with mixtures of Asp and 1,6-hexamethylenediamine (HA) at different ratios, mixed anion/cation exchanger supports were built. Only if HA greatly exceed Asp in the support, immobilization did not work. While only using the Asp-agarose support immobilized enzyme molecules were only ionically adsorbed after glutaraldehyde treatment (visualized in SDS-PAGE analysis), the mixed supports gave covalent immobilization. The glutaraldehyde modification of these biocatalysts permitted to establish covalent bonds with the support, and this was more effective when using higher amounts of HA in the support. When around 60 % of the groups in the support were HA, the treatment with glutaraldehyde fully suppressed enzyme release from the support after boiling in SDS. The glutaraldehyde treated biocatalysts were more stable than just the adsorbed enzymes or the enzyme adsorbed only on Asp supports and then treated with glutaraldehyde (the optimal biocatalyst retained 90 % of the initial activity while the just adsorbed ficin retained 50 % of the initial activity). This strategy can be utilized to immobilize other proteins with high isoelectric points following this immobilization strategy.
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The present study aims to analyze the interaction between Rhodotorula toruloides and magnetic nanoparticles and evaluate their effect on carotenoid production. The manganese ferrite nanoparticles were synthesized without chitosan (MnFe2O4) and chitosan coating (MnFe2O4-CS) by the co-precipitation method assisted by hydrothermal treatment. XRD (X-ray diffraction), Magnetometry, Dynamic Light Scattering (DLS) and FTIR (Fourier-Transform Infrared Spectroscopy), are used to characterize the magnetic nanoparticles. The crystallite size of MnFe2O4 was 16 nm for MnFe2O4 and 20 nm for MnFe2O4-CS. The magnetic saturation of MnFe2O4-CS was lower (39.6 ± 0.6 emu/g) than the same MnFe2O4 nanoparticles (42.7 ± 0.3 emu/g), which was attributed to the chitosan fraction presence. The MnFe2O4-CS FTIR spectra revealed the presence of the characteristic chitosan bands. DLS demonstrated that the average hydrodynamic diameters were 344 nm for MnFe2O4 and 167 nm for MnFe2O4-CS. A kinetic study of cell immobilization performed with their precipitation with a magnet demonstrated that interaction between magnetic nanoparticles and R. toruloides was characterized by an equilibrium time of 2 h. The adsorption isotherm models (Langmuir and Freundlich) were fitted to the experimental values. The trypan blue assay was used for cell viability assessment. The carotenoid production increased to 256.2 ± 6.1 µg/g dry mass at 2.0 mg/mL MnFe2O4-CS. The use of MnFe2O4-CS to stimulate carotenoid yeast production and the magnetic separation of biomass are promising nanobiotechnological alternatives. Magnetic cell immobilization is a perspective technique for obtaining cell metabolites.
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Carotenoides , Quitosana , Compostos Férricos , Compostos de Manganês , Rhodotorula , Rhodotorula/metabolismo , Quitosana/química , Compostos de Manganês/química , Compostos Férricos/química , Carotenoides/química , Nanopartículas de Magnetita/química , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
Brazil has one of the greatest biodiversities on the planet, where various crops play a strategic role in the country's economy. Among the highly appreciated biomasses is babassu, whose oil extraction generates residual babassu mesocarp (BM), which still needs new strategies for valorization. This work aimed to use BM as a support for the immobilization of Thermomyces lanuginosus lipase (TLL) in an 8.83â mL packed-bed reactor, followed by its application as a biocatalyst for the synthesis of hexyl laurate in an integrated process. Initially, the percolation of a solution containing 5â mg of TLL at 25 °C and flows ranging from 1.767 to 0.074â mL min-1 was investigated, where at the lowest flow rate tested (residence time of 2â h), it was possible to obtain an immobilized derivative with hydrolytic activity of 504.7 U g-1 and 31.7 % of recovered activity. Subsequent studies of treatment with n-hexane, as well as the effect of temperature on the immobilization process, were able to improve the activities of the final biocatalyst BM-TLLF, achieving a final hydrolysis activity of 7023â U g-1 and esterification activity of 430â U â g-1 against 142â U g-1 and 113.5â U g-1 respectively presented by the commercial TLIM biocatalyst. Desorption studies showed that the TL IM has 18â mg of protein per gram of support, compared to 4.92â mg presented by BM-TLL. Both biocatalysts were applied to synthesize hexyl laurate, achieving 98 % conversion at 40 °C within 2â h. Notably, BM-TLLF displayed exceptional recyclability, maintaining catalytic efficiency over 12 cycles. This reflects a productivity of 180â mg of product â h-1 U-1 of the enzyme, surpassing 46â mg h-1â U-1 obtained for TLIM. These results demonstrate the efficacy of continuous flow technology in creating a competitive and integrated process offering an exciting alternative for the valorization of residual lignocellulosic biomass.
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In light of the growing demand for novel biocatalysts and enzyme production methods, this study aimed to evaluate the potential of Aspergillus tubingensis for producing lipase under submerged culture investigating the influence of culture time and inducer treatment. Moreover, this study also investigated conditions for the immobilization of A. tubingensis lipase by physical adsorption on styrene-divinylbenzene beads (Diaion HP-20), for these conditions to be applied to an alternative immobilization system with a packed-bed reactor. Furthermore, A. tubingensis lipase and its immobilized derivative were characterized in terms of their optimal ranges of pH and temperature. A. tubingensis was shown to be a good producer of lipase, obviating the need for inducer addition. The enzyme extract had a hydrolytic activity of 23 U mL-1 and achieved better performance in the pH range of 7.5 to 9.0 and in the temperature range of 20 to 50 °C. The proposed immobilization system was effective, yielding an immobilized derivative with enhanced hydrolytic activity (35 U g-1), optimum activity over a broader pH range (5.6 to 8.4), and increased tolerance to high temperatures (40 to 60 â). This research represents a first step toward lipase production from A. tubingensis under a submerged culture and the development of an alternative immobilization system with a packed-bed reactor. The proposed system holds promise for saving time and resources in future industrial applications.
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Reatores Biológicos , Enzimas Imobilizadas , Lipase , Lipase/química , Lipase/metabolismo , Enzimas Imobilizadas/química , Enzimas Imobilizadas/metabolismo , Adsorção , Concentração de Íons de Hidrogênio , Aspergillus/enzimologia , Proteínas Fúngicas/química , TemperaturaRESUMO
Thermolysin (TLN) is a microbial highly-priced thermostable metallo-endoprotease with complementary substrate specificity to those of proteases widely used in science and industry for protein digestion and milk-clotting. This study is the first to immobilize TLN on aminated superparamagnetic nanoparticles (Fe3O4@silica-NH2) aiming for higher stability, recoverability, reusability, and applicability in proteolysis and as a microbial rennet-like milk-clotting enzyme. The nanobiocatalyst developed (Fe3O4@silica-TLN) displays hydrolytic activity on a synthetic TLN substrate and, apparently, was fully recovered from reaction media by magnetic decantation. More importantly, Fe3O4@silica-TLN retains TLN catalytic properties in the presence of calcium ions even after exposure to 60 °C for 48 h, storage at 4 °C for 80 days and room temperature for 42 days, use in proteolyses, and in milk-clotting for up to 11 cycles. Its proteolytic activity on bovine milk casein in 24 h furnished 84 peptides, of which 29 are potentially bioactive. Also, Fe3O4@silica-TLN catalyzed the digestion of bovine serum albumin. In conclusion, Fe3O4@silica-TLN showed to be a new, less autolytic, thermostable, non-toxic, magnetically-separable, and reusable nanobiocatalyst with highly attractive properties for both science (peptide/protein chemistry and structure, proteomic studies, and the search for new bioactive peptides) and food industry (cheese manufacture).
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Enzimas Imobilizadas , Leite , Proteólise , Dióxido de Silício , Termolisina , Dióxido de Silício/química , Animais , Leite/química , Enzimas Imobilizadas/química , Enzimas Imobilizadas/metabolismo , Termolisina/metabolismo , Termolisina/química , Biocatálise , Bovinos , Estabilidade Enzimática , Nanopartículas de Magnetita/químicaRESUMO
Corncob is an agro-residue rich in lignocellulosic material that can be used for the xylitol production, through its enzymatic conversion obtaining fermentable sugars and their subsequent fermentation. In light of the above, this study targeted the immobilization of Aspergillus labruscus xylanase and the use of the derivative to hydrolyze the corncob xylan for the obtainment of xylose, and its subsequent use for the production of xylitol. The extracellular xylanase was immobilized using different supports (sodium alginate, DEAE-Cellulose, DEAE-Sephadex and CM-Sephadex). Among all supports used, the best results were obtained with the DEAE-Cellulose derivative showing an efficiency of immobilization of 97-99%, yield of 93-95% and recovered activity of 81-100%. The sodium alginate derivative showed 3 cycles of reuse, with drop in activity of about 65% in the 3rd cycle using both CaCl2 and MnCl2 as crosslinkers. The best enzymatic activity for the DEAE-Cellulose derivative was observed at 55ºC and pH 5.0. This derivative presented reuse of 10 cycles using commercial xylan as substrate, and 4 cycles using corncob xylan. This derivative was used in an enzymatic reactor to hydrolyze corncob xylan, obtaining 2.7 mg/mL of xylose after 48 h of operation under optimal condition of temperature and pH. The xylose obtained from the corncob was fermented by Candida tropicalis for 96 h with consumption of 60%. The HPLC analyses indicated a production of 1.02 mg/mL of xylitol with 48 h of fermentation. In conclusion, this is the first report on the immobilization of the A. labrucus xylanase as an alternative for the obtainment of xylose from corncob xylan, and the subsequent production of xylitol.
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Microbially induced carbonate precipitation (MICP) immobilizes toxic metals and reduces their bioavailability in aqueous systems. However, its application in the treatment of acid mine drainage (AMD) is poorly understood. In this study, the genomes of Sporosarcina sp. UB5 and UB10 were sequenced. Urease, carbonic anhydrases, and metal resistance genes were identified and enzymatic assays were performed for their validation. The geochemical mechanism of precipitation in AMD was elucidated through geo-mineralogical analysis. Sporosarcina sp. UB5 was shown to be a new genomospecies, with an average nucleotide identity < 95 % (ANI) and DNA-DNA hybridization < 70 % (DDH) whereas UB10 is close to S. pasteurii. UB5 contained two urease operons, whereas only one was identified in UB10. The ureolytic activities of UB5 and UB10 were 122.67 ± 15.74 and 131.70 ± 14.35 mM NH4+ min-1, respectively. Both strains feature several carbonic anhydrases of the α, ß, or γ families, which catalyzed the precipitation of CaCO3. Only Sporosarcina sp. UB5 was able to immobilize metals and neutralize AMD. Geo-mineralogical analyses revealed that UB5 directly immobilized Fe (1-23 %), Mn (0.65-1.33 %) and Zn (0.8-3 %) in AMD via MICP and indirectly through adsorption to calcite and binding to bacterial cell walls. The MICP-treated AMD exhibited high removal rates (>67 %) for Ag, Al, As, Ca, Cd, Co, Cu, Fe, Mn, Pb, and Zn, and a removal rate of 15 % for Mg. This study provides new insights into the MICP process and its applications to AMD treatment using autochthonous strains.
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Mineração , Sporosarcina , Urease , Sporosarcina/genética , Sporosarcina/metabolismo , Urease/metabolismo , Precipitação Química , Carbonatos/química , Anidrases Carbônicas/metabolismo , Anidrases Carbônicas/genética , Poluentes Químicos da Água/metabolismo , Poluentes Químicos da Água/químicaRESUMO
Background: Whole-body vibration (WBV) is used to enhance physical performance in sports and rehabilitation. The present study analyzed the effects of remobilization with WBV on the soleus muscle of Wistar rats. Methods: Twenty-eight animals were separated into four experimental groups (n = 7): CON (control); IM (immobilized); FR (immobilization and free remobilization); and WBV (immobilization and remobilization with WBV). The immobilization of the pelvic limb was carried out according to the standard protocol using a plaster cast for 15 days. For remobilization with WBV, a Frequency of 60 Hz was applied for 10 min, five days a week, for two weeks. After the remobilization period, the animals were euthanized, and the right soleus muscle was dissected followed by processing for histomorphometric analysis and immunolocalization of Aquaporin 1 (AQP1). Results: We observed a reduced larger diameter in IM compared to CON, with restored values in WBV. For the estimation of connective tissue, a significant increase was observed in the immobilized groups, while a reduction was noted in the remobilized groups. AQP1 expression decreased significantly in IM and increased in WBV. Conclusion: Immobilization caused morphofunctional damage to the soleus muscle, and remobilization with WBV is efficient and offers advantages over free remobilization.
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Aquaporina 1 , Músculo Esquelético , Ratos Wistar , Vibração , Animais , Aquaporina 1/metabolismo , Músculo Esquelético/metabolismo , Músculo Esquelético/fisiologia , Ratos , Masculino , Imobilização/métodosRESUMO
Only a few halophilic archaea producing carboxylesterases have been reported. The limited research on biocatalytic characteristics of archaeal esterases is primarily due to their very low production in native organisms. A gene encoding carboxylesterase from Halobacterium salinarum NRC-1 was cloned and successfully expressed in Haloferax volcanii. The recombinant carboxylesterase (rHsEst) was purified by affinity chromatography with a yield of 81%, and its molecular weight was estimated by SDS-PAGE (33 kDa). The best kinetic parameters of rHsEst were achieved using p-nitrophenyl valerate as substrate (KM = 78 µM, kcat = 0.67 s-1). rHsEst exhibited great stability to most metal ions tested and some solvents (diethyl ether, n-hexane, n-heptane). Purified rHsEst was effectively immobilized using Celite 545. Esterase activities of rHsEst were confirmed by substrate specificity studies. The presence of a serine residue in rHsEst active site was revealed through inhibition with PMSF. The pH for optimal activity of free rHsEst was 8, while for immobilized rHsEst, maximal activity was at a pH range between 8 to 10. Immobilization of rHsEst increased its thermostability, halophilicity and protection against inhibitors such as EDTA, BME and PMSF. Remarkably, immobilized rHsEst was stable and active in NaCl concentrations as high as 5M. These biochemical characteristics of immobilized rHsEst reveal its potential as a biocatalyst for industrial applications.
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Carboxilesterase , Clonagem Molecular , Halobacterium salinarum , Proteínas Recombinantes , Carboxilesterase/genética , Carboxilesterase/metabolismo , Carboxilesterase/química , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Especificidade por Substrato , Halobacterium salinarum/enzimologia , Halobacterium salinarum/genética , Enzimas Imobilizadas/metabolismo , Enzimas Imobilizadas/química , Enzimas Imobilizadas/genética , Concentração de Íons de Hidrogênio , Cinética , Estabilidade Enzimática , Proteínas Arqueais/genética , Proteínas Arqueais/química , Proteínas Arqueais/metabolismo , TemperaturaRESUMO
Coastal wetlands are known for their diverse ecosystems, yet their soil characteristics are often misunderstood and thought to be monotonous. These soils are frequently subjected to saline water saturation, leading to unique soil processes. However, the combination and intensity of these processes can vary considerably across different ecosystems. In this study, we hypothesize that these diverse soil processes not only govern the geochemical conditions in coastal ecosystems but also influence their ability to deliver ecosystem services. To test this hypothesis, we conducted soil analyses in mangroves, seagrass meadows, and hypersaline tidal flats along the Brazilian coast. We used key soil properties as indicators of soil processes and developed a conceptual model linking soil processes and soil-related ecosystem services in these environments. Under more anoxic conditions, the intense soil organic matter accumulation and sulfidization processes in mangroves evidence their significance in terms of climate regulation through organic carbon sequestration and contaminants immobilization. Similarly, pronounced sulfidization in seagrasses underscores their ability to immobilize contaminants. In contrast, hypersaline tidal flats soils exhibit increased intensities of salinization and calcification processes, leading to a high capacity for accumulating inorganic carbon as secondary carbonates (CaCO3), underscoring their role in climate regulation through inorganic carbon sequestration. Our findings show that contrary to previously thought coastal wetlands are far from monotonous, exhibiting significant variations in the types and intensities of soil processes, which in turn influence their capacity to deliver ecosystem services. This understanding is pivotal for guiding effective management strategies to enhance ecosystem services in coastal wetlands.
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Solo , Áreas Alagadas , Solo/química , Brasil , Ecossistema , SalinidadeRESUMO
Although enzymes have been used for thousands of years, their application in industrial processes has gained importance since the 20th century due to technological and scientific advances in several areas, including biochemistry [...].
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In this article, the synthesis of antioxidant peptides in the enzymatic hydrolysis of caprine casein was analyzed at three different time points (60 min, 90 min, and 120 min) using immobilized pepsin on activated and modified carbon (AC, ACF, ACG 50, ACG 100). The immobilization assays revealed a reduction in the biocatalysts' activity compared to the free enzyme. Among the modified ones, ACG 50 exhibited greater activity and better efficiency for reuse cycles, with superior values after 60 min and 90 min. Peptide synthesis was observed under all studied conditions. Analyses (DPPH, ß-carotene/linoleic acid, FRAP) confirmed the antioxidant potential of the peptides generated by the immobilized enzyme. However, the immobilized enzyme in ACG 50 and ACG 100, combined with longer hydrolysis times, allowed the formation of peptides with an antioxidant capacity greater than or equivalent to those generated by the free enzyme, despite reduced enzymatic activity.
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Antioxidantes , Caseínas , Enzimas Imobilizadas , Glutaral , Cabras , Iridoides , Pepsina A , Peptídeos , Antioxidantes/química , Enzimas Imobilizadas/química , Enzimas Imobilizadas/metabolismo , Caseínas/química , Animais , Pepsina A/metabolismo , Pepsina A/química , Glutaral/química , Peptídeos/química , Iridoides/química , Hidrólise , Carvão Vegetal/químicaRESUMO
Objetivo: Extrair evidências científicas na literatura acerca dos fatores que podem causar imobilidade ou síndrome da imobilidade no idoso e possíveis intervenções. Métodos: Estudo do tipo revisão integrativa da literatura, realizado durante os meses de dezembro de 2019 a julho de 2020 por meio de buscas de evidências nas seguintes bases de dados: Medline, Scielo, Lilacs, Pubmed, Cinahl, Web of Science, Cochrane/Librany, Ebsco, Embase e Scorpus. Resultados: Foram identificadas 1656 produções, sendo selecionados dez estudos para a elaboração desta revisão. A maior parte das investigações foi produzida no Brasil, e as demais na Espanha, em Portugal, na Austrália e nos Estados Unidos. Os estudos evidenciaram que as causas da imobilidade no idoso são multifatoriais, que requerem intervenções específicas e adequadas, visando prevenir a imobilidade e a síndrome da imobilidade, suas consequências e melhorar a qualidade de vida dos idosos. Conclusão: Ressalta-se a necessidade de construir um instrumento que possibilite avaliar a síndrome da imobilidade no idoso, a fim de direcionar o cuidado individualizado e específico, assim como, promover a qualificação de profissionais na área de Gerontologia. (AU)
Objective: To extract scientific evidence in the literature about the factors that can cause immobility or immobility syndrome in the elderly and possible interventions. Methods: Study of the integrative literature review type performed from evidence searches in the databases: Medline, Scielo, Lilacs, Pubmed, Cinahl, Web of Science, Cochrane/Librany, Ebsco, Embase and Scorpus, during the months of December from 2019 to July 2020. Results: 1,656 productions were identified, and 10 studies were selected for the preparation of this review. Most of the investigations were produced in Brazil and the rest in Spain, Portugal, Australia and the United States. Studies have shown that the causes that compromise mobility in the elderly are multifactorial; indicate the need for specific and appropriate interventions, aimed at preventing immobility and immobility syndrome, and their consequences, with a view to improving the quality of life of the elderly. Conclusion: It points to the need to build an instrument to assess the immobility syndrome in the elderly, in order to guide individualized and specific care, as well as the importance of training professionals in the field of gerontology. (AU)
Objetivo: Extraer evidencia científica enla literatura sobre losfactores que pueden provocar inmovilidad o síndrome de inmovilidad em el anciano y posibles intervenciones. Métodos: Estudio del tipo de revisión integrativa de la literatura realizada a partir de búsquedas de evidencia em las bases de datos: Medline, Scielo, Lilacs, Pubmed, Cinahl, Web of Science, Cochrane / Librany, Ebsco, Embase y Scorpus, durante los meses de diciembre de 2019 a julio. 2020. Resultados: se identificaron 1.656 producciones y se seleccionaron 10 estudios para laelaboración de esta revisión. La mayoría de lasinvestigaciones se produjeronen Brasil y el resto em España, Portugal, Australia y Estados Unidos. Los estudioshan demostrado que las causas que comprometenlamovilidad em losancianossonmultifactoriales; indicanlanecesidad de intervenciones específicas y adecuadas, dirigidas a prevenir lainmovilidad y el síndrome de inmovilidad, y sus consecuencias, con miras a mejorarlacalidad de vida de las personas mayores. Conclusión: Es necesario construir un instrumento para evaluar el síndrome de inmovilidad em el anciano, com el fin de orientar cuidados individualizados y específicos, así como la importancia de formar professional esenel campo de la gerontología. (AU)
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Envelhecimento , Idoso , Idoso Fragilizado , Imobilização , Cuidados de EnfermagemRESUMO
In the quest for advanced and environmentally friendly solutions to address challenges in the field of wastewater treatment, the use of polymers such as sodium alginate (Na-Alg) in combination with immobilized microorganisms (IMs) stands out as a promising strategy. This study assesses the potential of Na-Alg in immobilizing microorganisms for wastewater treatment, emphasizing its effectiveness and relevance in environmental preservation through the use of IMs. Advances in IMs are examined, and the interactions between these microorganisms and Na-Alg as the immobilization support are highlighted. Additionally, models for studying the kinetic degradation of contaminants and the importance of oxygen supply to IMs are detailed. The combination of Na-Alg with IMs shows promise in the context of improving water quality, preserving ecological balance, and addressing climate change, but further research is required to overcome the identified challenges. Additional areas to explore are discussed, which are expected to contribute to the innovation of relevant systems.
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A laccase-based catalytic reactor was developed into a polydimethylsiloxane (PDMS) microfluidic device, allowing the degradation of different concentrations of the emergent pollutant, Bisphenol-A (BPA), at a rate similar to free enzyme. Among the immobilizing agents used, 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) was capable of immobilizing a more significant amount of the laccase enzyme in comparison to glutaraldehyde (GA), and the passive method (2989, 1537, and 1905 U/mL, respectively). The immobilized enzyme inside the microfluidic device could degrade 55 ppm of BPA at a reaction rate of 0.5309 U/mL*min with a contaminant initial concentration of 100 ppm at room temperature. In conclusion, the design of a microfluidic device and the immobilization of the laccase enzyme successfully allowed a high capacity of BPA degradation.
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Using advanced nanotechnology membranes has opened up new possibilities in the field of biomedicine, particularly for controlled drug delivery and especially for topical use. Bacterial cellulose membranes (BCM), particularly, have gained prominence owing to their distinctive attributes, including remarkable water retention, safety, biodegradability, and tunable gas exchange. However, they are aqueous matrices and, for this reason, of limited capacity for incorporation of apolar compounds. Cubosomes are lipid nanoparticles composed of a surfactant bicontinuous reverse cubic phase, which, owing to their bicontinuous structure, can incorporate both polar and apolar compounds. Therefore, these particles present a promising avenue for encapsulating and releasing drugs and biomolecules due to their superior entrapment efficiency. In this study, we aim to extend earlier investigations using polymeric hydrogels for cubosome immobilization, now using BCMs, a more resilient biocompatible matrix. Phytantriol cubosome-loaded BCMs were prepared by three distinct protocols: ex situ incorporation into wet BCMs, ex situ incorporation by swelling of dry BCMs, and an in situ process with the growth of BCMs in a sterile medium already containing cubosomes. Our investigation revealed that these methodologies ensured that cubosomes remained integral, uniformly distributed, and thoroughly dispersed within the membrane, as confirmed using Small-Angle X-ray Scattering (SAXS) and high-resolution confocal microscopy. The effective incorporation and sustained release of diclofenac were validated across the different BCMs and compared with hyaluronic acid (HA) hydrogel in our previous studies. Furthermore, the resistance against cubosome leaching from the three BCM and HA hydrogel samples was quantitatively evaluated and contrasted. We hope that the outcomes from this research will pave the way for innovative use of this platform in the incorporation and controlled release of varied active agents, amplifying the already multifaceted applicability of BCMs.
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Global market of food enzymes is held by pectinases, mostly sourced from filamentous fungi via submerged fermentation. Given the one-time use nature of enzymes to clarify juices and wines, there is a crucial need to explore alternatives for enzyme immobilization, enabling their reuse in food applications. In this research, an isolated fungal strain (Penicillium crustosum OR889307) was evaluated as a new potential pectinase producer in submerged fermentation. Additionally, the enzyme was immobilized in magnetic core-shell nanostructures for juice clarification. Findings revealed that Penicillium crustosum exhibited enzymatic activities higher than other Penicillium species, and pectinase production was enhanced with lemon peel as a cosubstrate in submerged fermentation. The enzyme production (548.93 U/mL) was optimized by response surface methodology, determining the optimal conditions at 35 °C and pH 6.0. Subsequently, the enzyme was covalently immobilized on synthesized magnetic core-shell nanoparticles. The immobilized enzyme exhibited superior stability at higher temperatures (50 °C) and acidic conditions (pH 4.5). Finally, the immobilized pectinases decreased 30 % the orange juice turbidity and maintained 84 % of the enzymatic activity after five consecutive cycles. In conclusion, Penicillium crustosum is a proven pectinase producer and these enzymes immobilized on functionalized nanoparticles improve the stability and reusability of pectinase for juice clarification.
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Nanopartículas , Penicillium , Poligalacturonase/química , Enzimas Imobilizadas/química , Penicillium/metabolismo , Temperatura , Fenômenos Magnéticos , Concentração de Íons de Hidrogênio , Estabilidade EnzimáticaRESUMO
This study presents an innovative method for synthesizing ß-amino carbonylated compounds, specifically 2-[phenyl(phenylamino)methyl] cyclohexanone, achieving high conversions and diastereomeric ratios. Using trypsin or α-chymotrypsin in both free and immobilized forms on titanate nanotubes (NtsTi), synthesized through alkaline hydrothermal methods, successful immobilization yields were attained. Notably, α-chymotrypsin, when free, displayed a diastereoselective synthesis of the anti-isomer with 97 % conversion and 16 : 84 (syn : anti) diastereomeric ratio, which slightly decreased upon immobilization on NtsTi. Trypsin, in its free form, exhibited diastereoselective recognition of the syn-isomer, while immobilization on NtsTi (trypsin/NtsTi) led to an inversion of diastereomeric ratio. Both trypsin/NtsTi and α-chymotrypsin/NtsTi demonstrated significant catalytic efficiency over five cycles. In conclusion, NtsTi serves as an effective support for trypsin and α-chymotrypsin immobilization, presenting promising prospects for diastereoselective synthesis and potential industrial applications. Furthermore, it offers promising prospects for the diastereoselective synthesis of 2-[phenyl(phenylamino)methyl] cyclohexanone through multicomponent Mannich reaction and future industrial application.
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Quimotripsina , Enzimas Imobilizadas , Nanotubos , Titânio , Tripsina , Titânio/química , Enzimas Imobilizadas/química , Enzimas Imobilizadas/metabolismo , Quimotripsina/química , Quimotripsina/metabolismo , Tripsina/metabolismo , Tripsina/química , Nanotubos/química , Estereoisomerismo , Biocatálise , Cicloexanonas/químicaRESUMO
Laccases are polyphenol oxidase enzymes and form the enzyme complex known for their role in wood decomposition and lignin degradation. The present study aimed to systematically review the state-of-the-art trends in scientific publications on laccase enzymes of the last 10 years. The main aspects checked included the laccase-producing fungal genera, the conditions of fungal growth and laccase production, the methods of immobilization, and potential applications of laccase. After applying the systematic search method 177 articles were selected to compound the final database. Although various fungi produce laccase, most studies were Trametes and Pleurotus genera. The submerged fermentation (SmF) has been the most used, however, the use of solid-state fermentation (SSF) appeared as a promising technique to produce laccase when using agro-industrial residues as substrates. Studies on laccase immobilization showed the covalent bonding and entrapment methods were the most used, showing greater efficiency of immobilization and a high number of enzyme reuses. The main use of the laccase was in bioremediation, especially in the discoloration of dyes from the textile industry and the degradation of pharmaceutical waste. Implications and consequences of all these findings in biotechnology and environment, as well as the trends and gaps of laccase research were discussed.
Assuntos
Biotecnologia , Enzimas Imobilizadas , Lacase , Lacase/metabolismo , Lacase/biossíntese , Lacase/química , Biotecnologia/métodos , Enzimas Imobilizadas/metabolismo , Enzimas Imobilizadas/química , Biodegradação Ambiental , Fungos/enzimologia , Fermentação , Proteínas Fúngicas/química , Proteínas Fúngicas/metabolismo , Corantes/metabolismo , Corantes/química , Pleurotus/enzimologiaRESUMO
The objective of this study was to obtain sufficient information on the thermal stabilization of a food-grade lipase from Thermomyces lanuginosus (TLL) using the immobilization technique. To do this, a new non-porous support was prepared via the sequential extraction of SiO2 from rice husks, followed by functionalization with (3-aminopropyl) triethoxysilane - 3-APTES (Amino-SiO2), and activation with glutaraldehyde - GA (GA-Amino-SiO2). We evaluated the influence of GA concentration, which varied from 0.25% v v-1 to 4% v v-1, on the immobilization parameters and enzyme thermal stabilization. The thermal inactivation parameters for both biocatalyst forms (soluble or immobilized TLL) were calculated by fitting a non-first-order enzyme inactivation kinetic model to the experimental data. According to the results, TLL was fully immobilized on the external support surface activated with different GA concentrations using an initial protein load of 5 mg g-1. A sharp decrease of hydrolytic activity was observed from 216.6 ± 12.4 U g-1 to 28.6 ± 0.9 U g-1 of after increasing the GA concentration from 0.25% v v-1 to 4.0% v v-1. The support that was prepared using a GA concentration at 0.5% v v-1 provided the highest stabilization of TLL - 31.6-times more stable than its soluble form at 60 °C. The estimations of the thermodynamic parameters, e.g., inactivation energy (Ed), enthalpy (ΔH#), entropy (ΔS#), and the Gibbs energy (ΔG#) values, confirmed the enzyme stabilization on the external support surface at temperatures ranging from 50 to 65 °C. These results show promising applications for this new heterogeneous biocatalyst in industrial processes given the high catalytic activity and thermal stability.