**Figure 2 here**

3.2 Kinetics of Ultrasound-assisted Enzymatic Hydrolysis for In Natura Seeds

The reactions were performed under the conditions of the optimal values obtained from Equation 4 (Power 70% for 3 minutes at initial room temperature, 1.8 B/O, and 0.25 C/S) at pH 4.2. The experimental data is shown in Figure 3 . As can be seen after 5 minutes the reaction reaches equilibrium at approximately 88% of FFA (0.75 mol kg-1). Besides, it has been verified that the ultrasound equipment heats up beyond usual when a time greater than 7 minutes is used, and, therefore, it is not possible to continue the reaction with the ultrasound. To evaluate the degree of enzyme deactivation and the potential of a higher yield of hydrolysis under the optimal conditions, experiments were carried out by continuing the reaction after 5 minutes in ultrasound in a refrigerated incubator with agitation (shaker) at room temperature and 140 rpm. After 30 minutes in the shaker, a yield of 89.96% was achieved for the hydrolysis reaction, and after 60 minutes, the yield was 91.57%. The absence of a significant increase in the hydrolysis yield, expose the deactivation of the catalyst after the 5 minutes under ultrasonic treatment, which is consistent with a result reported by Awadallak et al. (Awadallak et al., 2016), who studied the hydrolysis of soybean oil under ultrasound treatment, using a phospholipase as catalyst. The authors reported enzyme inacvation after 10 minutes when using 20% of ultrasound power (200 W) and after 5 minutes when using 50% of ultrasound power.
To better analyze the influence of employing ultrasound in the reaction, the same experiment, under the optimal conditions found throughEquation 4, was conducted in the refrigerated incubator (140 rpm) for 3 minutes. The yield obtained was 41.23 ± 2.62%, which is less than half of the yield obtained under 3 minutes of reaction under ultrasound treatment, 85.01%, demonstrating the high potential for the use of ultrasound in the enzymatic hydrolysis of Crambe oil using seeds in natura.
In Figure 3 , the concentration profiles of MAG, DAG, and TAG (in mol kg-1) are shown. The decrease in TAG (Figure 3 (d) ) concentration and increasing concentration of FFA (Figure 3 (a) ) are the expected behavior and can be seen. DAG (Figure 3 (c) ) and MAG (Figure 3 (b) ), behave in the usual way for intermediate compounds of a reaction. All the concentration profiles obey the principle of mass conservation (constant , i = TAG , DAG , MAG ,GL , H2O e FFA ). Additionally, inFigure 3, the model results for the concentration profiles of glycerol (Figure 3 (e) ) and water (Figure 3 (f) ) are shown. The value of 88% (≈ 0.75 mol kg-1) was reached after the equilibrium of the enzymatic hydrolysis of the Crambe oil in 5 min.
In Table 2 are presented the values of the estimated parameters from the fitting of the model, based on the Ping Pong Bi Bi (PPBB) kinetic mechanism, to the experimental kinetic data. The simulated and experimental curves are shown in Figure 3 . The coefficient of determination (R²) values were 1.00, 0.95, 0.88, and 0.99 for TAG, DAG, MAG, and FFA experimental data, respectively, which demonstrate that the kinetic model satisfactorily represented the set of experimental data.