Empleo de un extracto acuoso de ureasa obtenido a partir de la soya en la determinación de urea en sangre
Fecha
2016-06-25
Autores
Boffill Morrell, Taimi de la Caridad
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Editor
Universidad Central “Marta Abreu” de Las Villas. Facultad de Química Farmacia. Departamento de Licenciatura en Química
Resumen
En este trabajo se desarrolló la extracción de la ureasa presente en el frijol de soya, con el empleo de cuatro técnicas (Método de Lorenc, Método de Schosinsky, Método salting- in y Método de Donal y Van Slyke). A partir de una evaluación cualitativa y cuantitativa de la actividad enzimática y tomando como referencia los valores reportados por la literatura, se concluyó que los mejores resultados fueron los ofrecidos por el extracto de Lorenc. Se estudió la cinética a la reacción con la enzima ureasa del extracto obteniéndose los parámetros: Vmáx igual 3.125 mmol/L y Km igual 1.67*10-1 mol/L a una temperatura de 40 oC y pH igual a 7. Se determinó cuantitativamente la actividad para dicho extracto y se comparó con la actividad de la enzima ureasa comercial, procediéndose a aplicar el extracto a muestras de plasma sanguíneo, no presentándose diferencias significativas entre ambas enzimas para ambos ensayos. La caracterización del extracto mediante la determinación de proteínas totales por el método de Biuret mostró una concentración de albúmina de 8.67 mg/L y 20.83 mg/L de globulina. El estudio de estabilidad reveló que el extracto es estable por 7 días.
In this work the extraction of the present urease was developed in the soybean, with the employment of four techniques (Method of Lorenc, Method of Schosinsky, Method salting - in and Method of Donal and Van Slyke). Starting from a qualitative and quantitative evaluation of the enzymatic activity and taking like reference the values reported by the literature, we concluded that the best results were those offered by the extract of Lorenc. The kinetics was studied to the reaction with the enzyme urease of the extract being obtained the parameters: Vmáx same 3.125 mmol/L and same Km 1.67*10-1 mol/L to a temperature of 40 oC and pH similar to 7. It was determined the activity quantitatively for this extract and it was compared with the activity of the enzyme commercial urease, being proceeded to apply the extract to samples of sanguine plasm, not presenting differs significant among both enzymes for both rehearsals. The characterization of the extract by means of the determination of total proteins for the method of Biuret showed a concentration of albumin of 8.67 mg/L and 20.83 mg/L of globulin. The study of stability revealed that the extract is stable for 7 days.
In this work the extraction of the present urease was developed in the soybean, with the employment of four techniques (Method of Lorenc, Method of Schosinsky, Method salting - in and Method of Donal and Van Slyke). Starting from a qualitative and quantitative evaluation of the enzymatic activity and taking like reference the values reported by the literature, we concluded that the best results were those offered by the extract of Lorenc. The kinetics was studied to the reaction with the enzyme urease of the extract being obtained the parameters: Vmáx same 3.125 mmol/L and same Km 1.67*10-1 mol/L to a temperature of 40 oC and pH similar to 7. It was determined the activity quantitatively for this extract and it was compared with the activity of the enzyme commercial urease, being proceeded to apply the extract to samples of sanguine plasm, not presenting differs significant among both enzymes for both rehearsals. The characterization of the extract by means of the determination of total proteins for the method of Biuret showed a concentration of albumin of 8.67 mg/L and 20.83 mg/L of globulin. The study of stability revealed that the extract is stable for 7 days.
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Palabras clave
Frijol de Soya, Enzima Ureasa