Bioconjugation as a smart immobilization approach for α-amylase enzyme using stimuli-responsive Eudragit-L100 polymer: a robust biocatalyst for applications in pharmaceutical industry

Figures & data

Figure 1. Effect of polymer concentration on immobilization yield (%). Amylolytic activity was assayed under standard assay conditions.

Figure 2. Effect of enzyme concentration on immobilization yield (%). Amylolytic activity was assayed under standard assay conditions.

Figure 3. Effect of pH on amylolytic activity of free and immobilized α-amylase (E-AC).

Figure 4. Effect of temperature on amylolytic activity of free and immobilized α-amylase (E-AC) at various temperatures from 20–80 °C.

Table 1. Kinetic parameters (K_m, V_max) and amylolytic efficiency of free and bio-conjugated α-amylase enzyme (E-AC).

Enzyme sample	Km (mg/ml)	Vmax (mg/ml/min)	Amylolytic efficiency (Vmax/Km)
Free α-amylase	2.87	8.35	2.9
E-AC	3.15	8.98	2.85

Figure 5. Reusability of E-AC in repeated amylolytic reaction. Residual activity of the immobilized α-amylase (in %) after six cycles of repeated use (initial activity was taken as 100%).

Table 2. Half-life times (t_1/2) for the free and immobilized (E-CA) α-amylase at different temperatures.

Storage temperature	Free	E-CA	Stabilization factor
5°C	100 d	680 d	6.8
25°C	6 d	45 d	7.5
37°C	1.5 d	14 d	9.3
60°C	0.6 d	8 h	11.6

Figure 6. Storage stability of free and immobilized α-amylase (E-AC). Residual amylolytic activity was determined at different time intervals under standard assay conditions upon storage at 5 and 25 °C.