Improved reuse and affinity of enzyme using immobilized amylase on alginate matrix

Abstract

Abstract Enzyme immobilizations were widely used to increase their shelf life which is essential for the world’s industries. Therefore, amylase immobilized using Na-alginate as a matrix is necessary optimized and characterized. The parameters measured in the optimization of immobilization are the determination of the concentration of sodium alginate and contact time. Characterizations were conducted to determine the optimum concentration of substrate, the value Vmax, Michaelis-Menten constant (KM), pH, temperature, incubation time, and test reuse. The process of immobilized amylase activity test was performed in a continuous flow system using a reactor, and its sugar levels were determined using the Dinitro Salisilat Method (DNS). The results reveal that the immobilized amylase commercial has optimum concentration of Na-alginate of 5% (w/v) and contact time of 90 minutes with an immobilization efficiency value of 43.02%. Furthermore, the immobilized amylase has optimum activity at substrate concentrations of 3.5% (w/v), pH 4, incubation temperature of 40 °C, and a reaction time of 20 minutes with the value of the activity of 2760.4 U / mL. KM value of free amylase and immobilized amylase row are 0.18 mM and 0.15 mM, repectively. The value of KM immobilized amylase is smaller than the free enzyme. It proves that the immobilized amylase has a high affinity for the substrate. The immobilized amylase can be used up to 12 times with a value of the residual activity of 56.7%.