RESUMO
Lentil (Lens culinaris) is a protein-rich legume consumed worldwide and it also has the potential to become an alternative source of protein ingredient for human nutrition. The aim of this study was to determine the best processing parameters for the whole grain protein wet extraction, as well as to analyze the techno-functional properties, and physical characteristics of the protein concentrate and its flour. It was also evaluated the application of the concentrate into a fish-like croquette. The processing route was carried out by alkaline extraction and acid precipitation of the proteins where the pH, stirring time and solute:solvent ratio were evaluated. The final dried protein concentrate presented 85% protein on dry basis and a mass yield of 14%. The results were reproducible when tested on a first scaling up test. For the techno-functional properties, solubility, water and oil retention capacities, emulsification and foaming capacities and stability, and gelling capacity were tested. As for the food application into fish-like croquettes, the lentil protein showed similar scores for sensory acceptance, flavor and texture when compared to a commercial clean-taste concentrate. The results observed in this study were compatible to other alternative pulse-protein ingredients on the market, positioning lentil protein as a promising alternative protein source to produce ingredients for the plant-based market.
Assuntos
Manipulação de Alimentos , Lens (Planta) , Lens (Planta)/química , Manipulação de Alimentos/métodos , Proteínas de Plantas , Humanos , Solubilidade , Paladar , Concentração de Íons de HidrogênioRESUMO
Correcting the processing of ΔF508-CFTR, the most common mutation in cystic fibrosis, is the major goal in the development of new therapies for this disease. Here, we determined whether ΔF508 could be rescued by a combination of small-molecule correctors, and identified the mechanism by which correctors rescue the trafficking mutant of cystic fibrosis transmembrane conductance regulator (CFTR). We transfected COS-7 cells with ΔF508, created HEK-293 stably expressing ΔF508, and utilized CFBE41o(-) cell lines stably transduced with ΔF508. As shown previously, ΔF508 expressed less protein, was unstable at physiological temperature, and rapidly degraded. When the cells were treated with the combination C18 + C4 the mature C-band was expressed at the cell surface. After treatment with C18 + C4, we saw a lower rate of protein disappearance after translation was stopped with cycloheximide. To understand how this rescue occurs, we evaluated the change in the binding of proteins involved in endoplasmic reticulum-associated degradation, such as Hsp27 (HspB1) and Hsp40 (DnaJ). We saw a dramatic reduction in binding to heat shock proteins 27 and 40 following combined corrector therapy. siRNA experiments confirmed that a reduction in Hsp27 or Hsp40 rescued CFTR in the ΔF508 mutant, but the rescue was not additive or synergistic with C4 + 18 treatment, indicating these correctors shared a common pathway for rescue involving a network of endoplasmic reticulum-associated degradation proteins.
Assuntos
Regulador de Condutância Transmembrana em Fibrose Cística/metabolismo , Proteínas de Choque Térmico HSP27/metabolismo , Proteínas de Choque Térmico HSP40/metabolismo , Animais , Células COS , Chlorocebus aethiops , Regulador de Condutância Transmembrana em Fibrose Cística/genética , Células HEK293 , Humanos , Mutação , Ligação Proteica , TemperaturaRESUMO
In several plant tissues, programmed cell death (PCD) is mediated by the combined action of cysteine peptidases, namely KDEL-tailed cysteine peptidases (KDEL-CysEP) and vacuolar processing enzymes (VPE). Here, we performed a search of the draft genome of Jatropha curcas L. (Euphorbiaceae) and identified 2 genes for KDEL-CysEP (Jc-CysEP1 and Jc-CysEP2) and 3 genes for VPE (Jc-ßVPE, Jc-γVPE and Jc-δVPE) and determined the expression patterns of these genes by RT-qPCR in integument and cellular endosperm of seeds collected at seven different developmental stages. We were able to demonstrate that the expression of Jc-CysEP1, Jc-CysEP2, Jc-ßVPE and Jc-γVPE proceeded rapidly from Stage IV, with Jc-CysEP2 displaying the highest relative expression; expression of Jc-δVPE could not be detected in any of the tissues/developmental stages analyzed. Additionally, we showed that the expression pattern of these peptidases correlates with anatomical changes in integument and cellular endosperm, thus suggesting a role for both classes of peptidases in PCD and in protein processing, both of which occur simultaneously in each of these tissues.