RESUMEN
In this work, we describe an optimized procedure based on gradual hemolysis for the isolation of hemoglobin derived from bovine slaughterhouse erythrocytes in a membrane bioreactor. The membrane bioreactor system that provided high yields of hemoglobin (mainly oxyhemoglobin derivate) and its separation from the empty erythrocyte membranes (ghosts) was designed at a pilot scale. Ten different concentrations of hypotonic media were assessed from the aspect of the extent of hemolysis, hematocrit values of the erythrocyte suspensions, cell swelling, and membrane deformations induced by decreased salt concentration. Effective gradual osmotic hemolysis with an extent of hemolysis of 88% was performed using 35 mM Na-phosphate/NaCl buffer of pH 7.2-7.4. Under these conditions most of the cell membranes presented the appearance of the normal ghosts under phase contrast microscope. The hemoglobin purity of >80% was confirmed by SDS-PAGE. Kinetic studies showed that maximal concentration of hemoglobin was reached after 40 min, but the process cycle at which recovery of 83% was achieved lasted for 90 min. The dynamics of both steps, (1) transport through the membrane of erythrocytes during process of hemolysis and (2) transport through the reactor filters, were evaluated.
Asunto(s)
Reactores Biológicos , Eritrocitos/química , Hemoglobinas/aislamiento & purificación , Hemólisis , Animales , Transporte Biológico , Tampones (Química) , Bovinos , Electroforesis en Gel de Poliacrilamida , Membrana Eritrocítica/química , Hematócrito , Hemoglobinas/química , Soluciones Hipotónicas/química , Cinética , Microscopía de Contraste de Fase , Fragilidad Osmótica , Cloruro de Sodio/química , Factores de TiempoRESUMEN
BACKGROUND: Encapsulation of Thymus serpyllum L. aqueous extract within calcium alginate beads was studied in order to produce dosage formulations containing polyphenolic compounds. Electrostatic extrusion was applied for encapsulation of thyme aqueous extract in alginate gel beads. In addition to hydrogel beads, heat-dried and freeze-dried forms of beads were examined. METHODS: Encapsulation systems were examined and compared in order to choose the optimal one with respect to entrapment efficiency, preservation of antioxidant activity and thermal behaviour under heating conditions simulating the usual food processing. RESULTS: The beads obtained with approximately 2 mg g⻹ of gallic acid equivalents encapsulated in 0.015 g mL⻹ of alginate were spheres of a uniform size of about 730 µm. Encapsulation efficiency varied in the range 50-80% depending on the encapsulation method. Besides, the analysis reveals that the encapsulation process and the material used did not degrade the bioactive compounds, as the total antioxidant content remained unchanged. This was verified by Fourier transform infrared analysis, which proved the absence of chemical interactions between extracted compounds and alginate. Addition of a filler substance, such as sucrose and inulin, in the dried product reduced its collapse and roundness distortion during drying process. CONCLUSION: This study demonstrates the potential of using hydrogel material for encapsulation of plant poplyphenols to improve their functionality and stability in food products.