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The objective of this paper was to report the effect of ionic liquids (ILs) in the elaboration of nanofibers of cellulose bagasse from Agave tequilana Weber var. azul by the electrospinning method. The ILs used were 1-butyl-3-methylimidazolium chloride (BMIMCl), and DMSO was added as co-solvent. To observe the effect of ILs, this solvent was compared with the organic solvent TriFluorAcetic acid (TFA). The nanofibers were characterized by transmission electron microscopy (TEM), X-ray, Fourier transform-infrared using attenuated total reflection (FTIR-ATR) spectroscopy, and thermogravimetric analysis (TGA). TEM showed different diameters (ranging from 35 to 76 nm) of cellulose nanofibers with ILs (CN ILs). According to X-ray diffraction, a notable decrease of the crystalline structure of cellulose treated with ILs was observed, while FTIR-ATR showed two bands that exhibit the physical interaction between cellulose nanofibers and ILs. TGA revealed that CN ILs exhibit enhanced thermal properties due to low or null cellulose crystallinity. CN ILs showed better characteristics in all analyses than nanofibers elaborated with TFA organic solvent. Therefore, CN ILs provide new alternatives for cellulose bagasse. Due to their small particle size, CN ILs could have several applications, including in food, pharmaceutical, textile, and material areas, among others.
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This study describes the preparation and characterization of eggplant peel extract-loaded electrospun gelatin nanofiber and study of its in vitro release. Results obtained by scanning electron microscopy (SEM) and transmission electronic microscopy (TEM) micrograph revealed that eggplant peel extract-loaded electrospun gelatin nanofiber is in nanometric range with an average diameter 606.7 ± 184.5 and 643.6 ± 186.7 nm for 20 and 33.3 mg mL-1 of extract addition, respectively. Moreover, the incorporation of extract improved morphology by being smooth, homogeneous, and without account formation compared to nanofibers without extract (control). Fourier transform-infrared (FT-IR) spectra indicated that interaction exists between electrospun gelatin nanofiber and eggplant peel extract by hydrogen bond interactions, mainly. Electrospun gelatin nanofibers showed encapsulation efficiency greater than 90% of extract and a maximum release of 95 and 80% for the medium at pH 1.5 and 7.5, respectively. Therefore, the electrospinning technique is a good alternative for the conservation of bioactive compounds present in the eggplant peel through electrospun gelatin nanofiber.
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(1) Background: Lipases and esterases are important enzymes that share the α/ß hydrolase fold. The activity and cellular localization are important characteristics to understand the role of such enzymes in an organism. (2) Methods: Bioinformatic and biochemical tools were used to describe a new α/ß hydrolase from a Litopenaeus vannamei transcriptome (LvFHS for Family Serine Hydrolase). (3) Results: The enzyme was obtained by heterologous overexpression in Escherichia coli and showed hydrolytic activity towards short-chain lipid substrates and high affinity to long-chain lipid substrates. Anti-LvFHS antibodies were produced in rabbit that immunodetected the LvFSH enzyme in several shrimp tissues. (4) Conclusions: The protein obtained and analyzed was an α/ß hydrolase with esterase and lipase-type activity towards long-chain substrates up to 12 carbons; its immunodetection in shrimp tissues suggests that it has an intracellular localization, and predicted roles in energy mobilization and signal transduction.
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Hidrolases/metabolismo , Penaeidae/enzimologia , Sequência de Aminoácidos , Animais , Hidrolases/química , Hidrolases/genética , Espaço Intracelular/metabolismo , Modelos Moleculares , Penaeidae/citologia , Estrutura Secundária de Proteína , Serina/metabolismo , Transdução de SinaisRESUMO
Freezing conditions affect fish muscle protein functionality due to its denaturation/aggregation. However, jumbo squid (Dosidicus gigas) muscle protein functionality remains stable even after freezing, probably due to the presence of low-molecular-mass compounds (LMMC) as cryoprotectants. Thus, water-soluble LMMC (<1 kDa) fraction obtained from jumbo squid muscle was evaluated by Fourier transform infrared spectrometry. From its spectra, total carbohydrates, free monosaccharides, free amino acids and ammonium chloride were determined. Cryoprotectant capacity and protein cryostability conferred by LMMC were investigated by differential scanning calorimetry. Fraction partial characterization showed that the main components are free amino acids (18.84 mg/g), carbohydrates (67.1 µg/mg) such as monosaccharides (51.1 µg/mg of glucose, fucose and arabinose in total) and ammonium chloride (220.4 µg/mg). Arginine, sarcosine and taurine were the main amino acids in the fraction. LMMC, at the mass fraction present in jumbo squid muscle, lowered the water freezing point to -1.2 °C, inhibiting recrystallization at 0.66 °C. Significant myofibrillar protein stabilization by LMMC was observed after a freeze-thaw cycle compared to control (muscle after extraction of LMMC), proving the effectiveness on jumbo squid protein muscle cryo- stability. Osmolytes in LMMC fraction inhibited protein denaturation/aggregation and ice recrystallization, maintaining the muscle structure stable under freezing conditions. LMMC conferred protein cryostability even at the very low mass fraction in the muscle.
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BACKGROUND: The secondary structure of a protein determines its functional properties, such as its gelling capacity. The α-helix and ß-sheet comprise its main structures. Myofibrillar proteins from jumbo squid are composed mainly of the actomyosin-paramyosin complex; this complex contains a high percentage of α-helix, because actin, paramyosin, and myosin constitute 30%, 100%, and 55% of the α-helix, respectively. It is important to elucidate the role of the secondary structures in the gelation of giant squid proteins as they form gel. The role of the secondary structures in the gelation of giant squid proteins is therefore very important. For this reason, the objective of this work was to evaluate the effect of temperature on the structural behavior of actomyosin-paramyosin isolate (API) from Dosidicus gigas. RESULTS: The unfolding of the API system, which is composed of the actomyosin-paramyosin complex, was clarified by studying surface hydrophobicity and viscosity. Three characteristic peaks were found, associated with myosin, paramyosin, and actin. Infrared and circular dichroism corroborated the view that API undergoes major structural changes, because it proceeds from mostly an α-helix structure to 100% ß-sheet. CONCLUSION: The structural rearrangement favors gelation by cross-linking, generating new protein-protein and water-protein interactions, which create a more stable structure compared to mantle proteins (MP). Likewise, the presence of sarcoplasmic and stromal proteins in D. gigas muscle prevents the unfolding of myofibrillar proteins, favoring gelation by agglomeration, decreasing the ability to trap water and thus its gelling capacity. © 2019 Society of Chemical Industry.
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Actomiosina/química , Decapodiformes/química , Alimentos Marinhos/análise , Tropomiosina/química , Animais , Interações Hidrofóbicas e Hidrofílicas , Miosinas/química , Estrutura Secundária de Proteína , Desdobramento de Proteína , TemperaturaRESUMO
BACKGROUND: Trypsin from fish species is considered as a cold-adapted enzyme that may find potential biotechnological applications. In this work, the recombinant expression, refolding and activation of Trypsin I (TryI) from Monterey sardine (Sardinops sagax caerulea) are reported. METHODS: TryI was overexpressed in Escherichia coli BL21 as a fusion protein of trypsinogen with thioredoxin. Refolding of trypsinogen I was achieved by dialysis of bacterial inclusion bodies with a recovery of 16.32 mg per liter of Luria broth medium. RESULTS: Before activation, the trypsinogen fusion protein did not show trypsin activity. Trypsinogen I was activated by adding 0.002 U of native TryI purified from the sardine pyloric caeca (nonrecombinant). The activated recombinant trypsin showed three times more activity than the nonrecombinant trypsin alone. CONCLUSION: The described protocol allowed obtaining sufficient amounts of recombinant TryI from Monterey sardine fish for further biochemical and biophysical characterization of its coldadaptation parameters.
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Escherichia coli , Proteínas de Peixes , Peixes/genética , Corpos de Inclusão , Redobramento de Proteína , Tripsina , Animais , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas de Peixes/biossíntese , Proteínas de Peixes/química , Proteínas de Peixes/genética , Proteínas de Peixes/isolamento & purificação , Corpos de Inclusão/química , Corpos de Inclusão/genética , Corpos de Inclusão/metabolismo , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Tripsina/biossíntese , Tripsina/química , Tripsina/genética , Tripsina/isolamento & purificaçãoRESUMO
In this study, cellulose of bagasse from Agave tequilana Weber var. azul was extracted to elaborate nanofibers by the electrospinning technique. Fiber characterization was performed using Transmission Electron Microscopy (TEM), x-ray, Fournier Transform-InfraRed (FT-IR) spectroscopy, and thermal analysis by Differential Scanning Calorimetry-Thermogravimetric Analysis (DSC-TGA). Different diameters (ranging from 54.57⯱â¯0.02 to 171⯱â¯0.01â¯nm) of nanofibers were obtained. Cellulose nanofibers were analyzed by means of x-ray diffraction, where we observed a total loss of crystallinity in comparison with the cellulose, while FT-IR spectroscopy revealed that the hemicellulose and lignin present in the agave bagasse were removed. Thermal analysis showed that nanofibers exhibit enhanced thermal properties, and the zeta potential value (-32.5â¯mV) demonstrated moderate stability in the sample. In conclusion, the nanofibers obtained provide other alternatives-of-use for this agro-industrial residue and could have potential in various industrial applications, among these encapsulation of bioactive compounds and reinforcing material, to mention a few.
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Cancer is the leading cause of morbidity and mortality worldwide. Therefore, the search for new and less aggressive treatments is currently the focus of the anticancer research. An attractive alternative for this purpose is the use of bioactive peptides from plants. Plants live everywhere on Earth, both on land and in water, and they are a major source of diverse molecules with pharmacological potential as antioxidant peptides. Hence, this review focuses on the importance of the antioxidant activity of terrestrial and aquatic plant peptides against cancer throughout several mechanisms. The influence of the antioxidant activity of peptides by different factors such as molecular weight and amino acid composition as a crucial factor for anticancer activity is also revised. Furthermore, the relation of antioxidant activity with anticancer property as well as safety and legal aspects of protein hydrolysates and bioactive peptides for their use in cancer treatments is discussed.
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Antineoplásicos/química , Antioxidantes/química , Neoplasias/tratamento farmacológico , Peptídeos/química , Plantas/química , Antineoplásicos/isolamento & purificação , Antineoplásicos/farmacologia , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Organismos Aquáticos , Humanos , Peptídeos/isolamento & purificação , Peptídeos/farmacologia , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Hidrolisados de Proteína/farmacologiaRESUMO
BACKGROUND: The giant squid (Dosidicus gigas) has been proposed as raw material to obtain myofibrillar protein concentrates. However, it has been observed that colloidal systems formed from squid proteins have limited stability. Therefore, the isolation and characterization of the actomyosin-paramyosin isolated (API) complex were performed, because they are the main proteins to which functionality has been attributed. RESULTS: Densitogram analysis revealed 45% of actin, 38% of myosin and 17% of paramyosin. The amino acid profile indicates a higher proportion of acidic amino acids, which gives a higher negative charge; this was supported by the zeta potential. Total sulfhydryl (TSH) content was lower compared with proteins of other aquatic species. CONCLUSION: The higher percentage of actin in relation to myosin, the presence of paramyosin, as well as the low content of sulfhydryl groups, could comprise the main causes of the low technological functional property of proteins from D. gigas mantle. © 2017 Society of Chemical Industry.
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Actomiosina/química , Decapodiformes/química , Tropomiosina/química , Actinas/química , Actinas/metabolismo , Actomiosina/metabolismo , Animais , Decapodiformes/metabolismo , Estabilidade Proteica , Alimentos Marinhos/análise , Tropomiosina/metabolismoRESUMO
This study aimed to establish the combined effect of aflatoxin B1 (AFB1) and fumonisin B1 (FB1) on wild Litopenaeus vannamei hepatopancreas alkaline phosphatase (AP) activity compared with that of farmed shrimp. AP activity in hepatopancreas extract was confirmed by several specific inhibitor assays. AP activity of wild shrimp was higher than that of farmed shrimp (p < 0.05). However, AP activity from both wild and farmed shrimp was inhibited when incubated with AFB1 and FB1. The greatest inhibition occurred when AP was incubated with a mixture of AFB1 and FB1. The IC50 for AFB1 on AP activity of wild and farmed shrimp hepatopancreases was 0.790 and 0.398 µg/mL, respectively. The IC50 of FB1 was 0.87 µg/mL for wild shrimp and 0.69 µg/mL for farmed shrimp. These results suggest that, at the mycotoxins concentrations used in the study, AP from farmed L. vannamei was sensitive to the presence of both mycotoxins; however, AP is more sensitive to the combination of AFB1 + FB1 suggesting a possible synergistic or potentiating inhibitory effect.