RESUMO
The objective of this research was to identify and characterize the encoded peptides present in nut storage proteins of Carya illinoinensis. It was found, through in silico prediction, proteomic analysis, and MS spectrometry, that bioactive peptides were mainly found in albumin and glutelin fractions. Glutelin was the major fraction with ~53% of the nut storage proteins containing at least 21 peptides with different putative biological activities, including antihypertensives, antioxidants, immunomodulators, protease inhibitors, and inhibitors of cell cycle progression in cancer cells. Data showed that using 50 µg/mL tryptic digests of enriched peptides obtained from nut glutelins is able to induce up to 19% of apoptosis in both HeLa and CasKi cervical cancer cells. To our knowledge, this is the first report that shows the potential value of the nut-encoded peptides to be considered as adjuvants in cancer therapies.
Assuntos
Antioxidantes/química , Carya/química , Peptídeos/química , Proteínas de Armazenamento de Sementes/genética , Antioxidantes/isolamento & purificação , Carya/genética , Glutens/química , Glutens/genética , Nozes/química , Nozes/genética , Peptídeos/genética , Peptídeos/isolamento & purificação , Inibidores de Proteases/química , Inibidores de Proteases/isolamento & purificação , Proteômica , Proteínas de Armazenamento de Sementes/química , Proteínas de Armazenamento de Sementes/isolamento & purificação , Sementes/químicaRESUMO
The presence of endocrine disruptors bisphenol-A, bisphenol-A-dimethacrylate, bisphenol-A-diglycidyl-ether, phthalic-acid, dibutyl-phthalate, diethyl-phthalate and dioctyl-phthalate was determined in vegetable cans, baby bottles and microwaveable containers from the Mexican market. Gas-Chromatography-Mass-Spectrometry was used for the identification and High-Performance-Liquid-Chromatography with UV/Visible light and fluorescence detectors was used for the quantification. Endocrine disruptors were found in all samples. PA and DOP were the substances most commonly found, and maximum concentrations were 9.549 and 0.664 µg/kg, respectively from a jalapeno peppers can. Bisphenol A, phthalic-acid, bisphenol-A-dimethacrylate, bisphenol-A-diglycidyl-ether, dioctyl-phtalate and dibutyl-phthalate were found in baby bottles and microwaveable containers.
Assuntos
Contaminação de Alimentos , Embalagem de Alimentos , Alimentos em Conserva/análise , Fenóis/análise , Ácidos Ftálicos/análise , Plásticos/química , Compostos Benzidrílicos , Alimentação com Mamadeira/instrumentação , Qualidade de Produtos para o Consumidor , Dibutilftalato/análise , Dibutilftalato/química , Dietilexilftalato/análise , Dietilexilftalato/química , Exposição Ambiental/análise , Exposição Ambiental/estatística & dados numéricos , México , Fenóis/química , Ácidos Ftálicos/químicaRESUMO
A synthetic human interferon gamma (hIFN-gamma) gene was fused to SP1 and SP3, two Sec-dependent artificial signal peptides to transport the hIFN-gamma to the periplasm of Escherichia coli BL21-SI. The processing efficiency of both SP1-hIFN-gamma and SP3-hIFN-gamma was dependent on the culture medium as well as the post-induction temperature. Both precursors were processed completely when cells were cultured using minimal medium and a post-induction temperature of 32.5 degrees C, and only the processed hIFN-gamma was detected. The SP3 signal peptide was more efficient than SP1 for the secretion of hIFN-gamma. Sixty percent of the total hIFN-gamma was secreted to the periplasm using the SP3 signal peptide and a post-induction temperature of 20 degrees C. Using Tris-sucrose-dithiothreitol (TSD) hypertonic buffer, the periplasmic soluble hINF-gamma was recovered with a purity of 85%.
Assuntos
Escherichia coli/metabolismo , Interferon gama/biossíntese , Periplasma/metabolismo , Sequência de Aminoácidos , Sequência de Bases , Escherichia coli/ultraestrutura , Humanos , Interferon gama/isolamento & purificação , Dados de Sequência Molecular , Sinais Direcionadores de Proteínas/genética , TemperaturaRESUMO
Cheese whey (CW) is the major subproduct from cheese manufacturing and it is considered as a waste pollutant since its high content of lactose. In this work a fermentation process for the production of penicillin acylase (PA) by a recombinant Escherichia coli and using CW as unique carbon source and inducer was developed. A design factorial 3(2) was used to evaluate the influence of independent variables (dissolved oxygen and CW concentration) on the ability of E. coli W3110/pPA102 to produce PA. Maximum specific PA activity of 781 U g(-1) was attained at 5 g L(-1) of CW and 3% dissolved oxygen. The results showed that CW can be used successfully as unique carbon source and inducer for the production of recombinant proteins using constructions driven by the lac promoter and this way reducing the discharges of that pollutant to the environment.