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Sustainable extraction processes based on alternative solvents to recover bioactive compounds of different raw materials have been highlighted as excellent alternatives to supply the needs of society towards a bioeconomy strategy. Little is known about the safety and biological effect of compounds extracted by these processes. In this work, carotenoids from Bactris gasipaes wastes obtained by an IL-based process were investigated in terms of safety, anti-inflammatory and, antioxidant activity in a high-fat-diet animal model on the kidney. Wistar rats were supplemented or not by carotenoids extracted with IL or VOS. The animals supplemented with carotenoids had lower weight than control and high-fat diets. In the animals supplemented with carotenoids, the group IL improved anti-inflammatory and antioxidant activity compared with carotenoids obtained by VOS. Also, the group HFD-VOS showed moderate-severe injuries on the kidney. Then, ILs could represent a novel tool for natural pigments safely applied to food industry.
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Cytochrome c (Cyt-c), a small mitochondrial electron transport heme protein, has been employed in bioelectrochemical and therapeutic applications. However, its potential as both a biosensor and anticancer drug is significantly impaired due to poor long-term and thermal stability. To overcome these drawbacks, we developed a site-specific PEGylation protocol for Cyt-c. The PEG derivative used was a 5 kDa mPEG-NHS, and a site-directed PEGylation at the lysine amino-acids was performed. The effects of the pH of the reaction media, molar ratio (Cyt-c:mPEG-NHS) and reaction time were evaluated. The best conditions were defined as pH 7, 1:25 Cyt-c:mPEG-NHS and 15 min reaction time, resulting in PEGylation yield of 45% for Cyt-c-PEG-4 and 34% for Cyt-c-PEG-8 (PEGylated cytochrome c with 4 and 8 PEG molecules, respectively). Circular dichroism spectra demonstrated that PEGylation did not cause significant changes to the secondary and tertiary structures of the Cyt-c. The long-term stability of native and PEGylated Cyt-c forms was also investigated in terms of peroxidative activity. The results demonstrated that both Cyt-c-PEG-4 and Cyt-c-PEG-8 were more stable, presenting higher half-life than unPEGylated protein. In particular, Cyt-c-PEG-8 presented great potential for biomedical applications, since it retained 30-40% more residual activity than Cyt-c over 60-days of storage, at both studied temperatures of 4 °C and 25 °C.
Assuntos
Citocromos c , Lisina , Dicroísmo Circular , Citocromos c/química , Lisina/química , Polietilenoglicóis/química , Polietilenoglicóis/metabolismo , Polietilenoglicóis/farmacologia , ProteínasRESUMO
Currently, the ability to produce several kinds of water-soluble red natural colorants makes the genus Talaromyces particularly important to the dye industry, which can be an alternative to the use of harmful synthetic colorants. In this study, colored compounds produced by Talaromyces amestolkiae were extracted, characterized chemically and the color stability of the fermented broth without any extraction procedure was further evaluated over pH variation. Five azaphilones compounds were detected by Ultrahigh Performance Liquid Chromatography-Mass Spectrometry system, all being complexes of the fatty acid amino-hexanedioic acid and azaphilone Monascus colorants. The color of the fermented broth was stable at a wide range of pH (3-9). Furthermore, T. amestolkiae colorants precipitated through hydrolysis of key chemical groups at extremely acidic (pH 1) and lose red color in extremely basic (pH 13) medium, showing negative halochromism. Nevertheless, these findings enhance the industrial relevance of azaphilone colorants produced by biotechnological process.
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Monascus , Talaromyces , Benzopiranos , Pigmentos Biológicos , Talaromyces/genéticaRESUMO
Biological drugs or biopharmaceuticals off patent open a large market for biosimilars and biobetters, follow-on biologics. Biobetters, in particular, are new drugs designed from existing ones with improved properties such as higher selectivity, stability, half-life and/or lower toxicity/immunogenicity. Glycosylation is one of the most used strategies to improve biological drugs, nonetheless bioconjugation is an additional alternative and refers to the covalent attachment of polymers to biological drugs. Extensive research on novel polymers is underway, nonetheless PEGylation is still the best alternative with the longest clinical track record. Innovative trends based on genetic engineering techniques such as fusion proteins and PASylation are also promising. In this review, all these alternatives wereexplored as well as current market trends, legislation and future perspectives.
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Produtos Biológicos , Medicamentos Biossimilares , Produtos Biológicos/farmacologia , Produtos Biológicos/normas , Medicamentos Biossimilares/farmacologia , Medicamentos Biossimilares/normas , Tratamento Farmacológico/tendências , Humanos , Engenharia Metabólica/métodos , Engenharia de Proteínas/métodos , Melhoria de QualidadeRESUMO
Ionic liquids (ILs) have been proposed as more efficient and sustainable solvents to replace volatile organic solvents (VOSs). However, the drawbacks associated with their use are still limiting the regular application of bioactive compounds obtained from the processes they mediate as food ingredients. It is true that the number of ILs approved by the Food and Drug Administration for food applications is still low and mainly focused on the ones from the quaternary ammonium family. However, this trend is changing, judging from the evidence that industries are surpassing overgeneralization about ILs (on price and toxicity) and starting to consider the potential and performance of ILs as solvents. Despite the examples of industries applying ILs in their processes, the use of bioactive compounds obtained from IL-based processes as ingredients in food formulations is still a big challenge. The positive influence of carotenoids on diseases associated or originating from the inflammatory scenario including, among others, obesity, is not new. Moreover, it is also well known that the poorest population worldwide does not have the recommended intake of carotenoids, especially those pro-vitaminic A. In an attempt to help answer this issue, dietary supplements containing adequate doses of natural carotenoids are expected to be the solution, or at least, part of the solution for a healthier life, but also, to reduce hunger. Thus, complete studies evaluating the toxicological potential and the real viability of adding these bioactive compounds in food formulations proving (or not!) their safety to consumers and handlers are highly demanded. This work proposes to investigate the potential of carotenoids extracted from Bactris gasipaes feedstocks mediated by an ethanolic solution of an imidazolium-based IL. Thus, male Wistar rats were randomized in six different groups, supplemented or not by carotenoids extracted by IL or VOS, and fed by control- and/or high-fat-diets (HFD). The adipose tissue-liver axis was studied as a model to investigate the influence of the carotenoids on the levels of inflammation and oxidative stress markers. The main results showed that animals supplemented with carotenoids extracted with IL displayed improvements in serum parameters, besides lower metabolic efficiency, and antioxidant response on the liver, even when fed with HFD. However, animals supplemented with carotenoids extracted by VOS showed higher levels of pro-inflammatory markers and huge oxidative stress on the liver.
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Tecido Adiposo/efeitos dos fármacos , Anti-Inflamatórios/farmacologia , Carotenoides/farmacologia , Inflamação/tratamento farmacológico , Líquidos Iônicos/química , Fígado/efeitos dos fármacos , Animais , Peso Corporal/efeitos dos fármacos , Carotenoides/química , Metabolismo Energético/efeitos dos fármacos , Masculino , Ratos , Ratos WistarRESUMO
This review will discuss, under the Circular Economy and Biorefinery concepts, the performance of the alternative solvents in the downstream process to recover natural pigments in a more sustainable way. Conventionally, pigments marketed on an industrial scale are produced through chemical synthesis by using petroleum derivatives as the main raw material. Also, the current production chain of the synthetic dyes is linear, with no solvent recycling and waste generation. Thus, the most promising processes of extraction and purification of natural pigments and strategies on the polishing of the solvents are here reviewed. In this review, the use of alternative solvents, namely, ionic liquids, eutectic solvents, aqueous solutions of surfactants, and edible oils, for recovering natural pigments was reviewed. Works discussing higher extraction yields and selectivity, while maintaining the stability of the target pigments, were reported. Also, a panorama between Sustainability and Circular Economy prospection was discussed for better comprehension of the main advances in the field. Behind the analysis of the works published so far on the theme, the most important lacunas to overcome in the next years on the field were pointed out and discussed. Also, the future trends and new perspectives to achieve the economic viability and sustainability of the processes using alternative solvents will be scrutinized.
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Líquidos Iônicos , Fenômenos Químicos , Corantes , Alimentos , SolventesRESUMO
Ionic liquids (ILs) are often claimed to be "environmentally friendly" compounds however, the knowledge of their potential toxicity towards different organisms and trophic levels is still limited, in particular when protic ionic liquids (PILs) are addressed. This study aims to evaluate the toxicity against various microorganisms and the biodegradability of four PILs namely, N-methyl-2-hydroxyethylammonium acetate, m-2-HEAA; N-methyl-2-hydroxyethylammonium propionate, m-2-HEAPr; N-methyl-2-hydroxyethylammonium butyrate, m-2-HEAB; and N-methyl-2-hydroxyethylammonium pentanoate, m-2-HEAP. The antimicrobial activity was determined against the two bacteria, Sthaplylococcus aureus ATCC-6533 and Escherichia coli CCT-0355; the yeast Candida albicans ATCC-76645; and the fungi Fusarium sp. LM03. The toxicity of all PILs was tested against the aquatic luminescent marine bacterium Vibrio fischeri using the Microtox(®) test. The impact of the PILs was also studied regarding their effect on lettuce seeds (Lactuta sativa). The biodegradability of these PILs was evaluated using the ratio between the biochemical oxygen demand (BOD) and the chemical oxygen demand (COD). The results show that, in general, the elongation of the alkyl chain tends to increase the negative impact of the PILs towards the organisms and biological systems under study. According to these results, m-2-HEAA and m-2-HEAP are the less and most toxic PILs studied in this work, respectively. Additionally, all the PILs have demonstrated low biodegradability.
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Líquidos Iônicos/toxicidade , Compostos de Amônio Quaternário/toxicidade , Aliivibrio fischeri/efeitos dos fármacos , Aliivibrio fischeri/metabolismo , Antibacterianos/química , Antibacterianos/metabolismo , Antibacterianos/toxicidade , Antifúngicos/química , Antifúngicos/metabolismo , Antifúngicos/toxicidade , Biodegradação Ambiental , Análise da Demanda Biológica de Oxigênio , Candida albicans/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Fusarium/efeitos dos fármacos , Líquidos Iônicos/química , Líquidos Iônicos/metabolismo , Lactuca/efeitos dos fármacos , Lactuca/crescimento & desenvolvimento , Luminescência , Compostos de Amônio Quaternário/química , Compostos de Amônio Quaternário/metabolismo , Staphylococcus aureus/efeitos dos fármacosRESUMO
A single-step selective separation of two food additives was investigated using alcohol-salt aqueous two-phase systems (ATPS). The selective partitioning of two of the most used additives from a processed food waste material, vanillin and l-ascorbic acid, was successfully accomplished. The results obtained prove that alcohol-salt ATPS can be easily applied as cheaper processes for the selective recovery of valuable chemical products from food wastes and other sources. As a first approach, the phase diagrams of ATPS composed of different alcohol+inorganic salt+water were determined at 298 (± 1)K and atmospheric pressure. The influence of methanol, ethanol, 1-propanol, and 2-propanol and K(3)PO(4), K(2)HPO(4) or KH(2)PO(4)/K(2)HPO(4) in the design of the phase diagrams was addressed. After the evaluation of the phase diagrams behaviour, the influence of the phase forming constituents was assessed towards the partition coefficients and recovery percentages of vanillin and l-ascorbic acid among the coexisting phases. Both model systems and real processed food waste materials were employed. Using these ATPS as partitioning systems it is possible to recover and separate vanillin, which migrates for the alcohol-rich phase, from l-ascorbic acid, which preferentially partitions for the salt-rich phase.