Soybean phospholipase D activity determination. A comparison of two methods

Abstract

RESUMENFosfolipasa D en soja. Comparación de la actividad determinada por dos métodos.Se evaluaron dos métodos para determinar la actividad de la fosfolipasa D en soja debido a que existe discrepancia entre los resultados publicados. Un método se basa en la extracción de la enzima de la harina resultante de la molienda del grano de soja entero, cuantificando la actividad sobre el extracto. En el otro método, la cuantificación se realiza sobre la harina del grano entero molido, sin extraer la enzima. En el método de extracción se optimizaron tanto el tiempo como el número de extracciones. Los mayores valores de actividad de la fosfolipasa D se obtuvieron por el método sin extracción de la enzima. Este método es más simple, exige menos tiempo de ejecución y las condiciones del medio en que actúa la fosfolipasa D se asemejan a las condiciones encontradas en la industria aceitera. PALABRAS-CLAVE: Actividad de la fosfolipasa D -Fosfátidos hidratables -Fosfátidos no hidratables -Fosfolipasa D -Soja SUMMARY Soybean phospholipase D activity determination. A comparison of two methods.Due to a discrepancy between previously published results, two methods to determine the soybean phospholipase D activity were evaluated. One method is based on the extraction of the enzyme from whole soybean flour, quantifying the enzyme activity on the extract. The other method quantifies the enzymatic activity on whole soybean flour without enzyme extraction. In the extraction-based-method, both the extraction time and the number of extractions were optimized. The highest phospholipase D activity values were obtained from the method without enzyme extraction. This method is less complex, requires less running-time and the conditions of the medium in which phospholipase D acts resemble the conditions found in the oil industry.

Isolation, purification and characterization of Phospholipase D from almond (Amygdalus spinosissima) from Ziarat, Balochistan

“…The (Fig. 1) shows that 1.5 hrs is the best time for high enzyme activity the obtained result is similar with that of a previously published study on soybean [11] and was not in agreement with the results of a published study on Streptomyces recemochromogenes which was 1 hr [19]. Hydrophobic affinity chromatography was used to purify PLD.…”

“…The sample was crushed with the help of pestle and mortar using liquid nitrogen and stored at 20℃. Extraction PLD was extracted according to Tosi et al, [11]. The sample was weighted 5.0g and mixed in 50ml of 0.1M sodium acetate buffer (pH 5.6).…”

“…By following Tosi et al [11], the activity of PLD was determined according to Bergmeyer & Gavehn, [12]. Supernatant (10ml) was transferred in an Erlenmeyer flask.…”