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

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%.

Preparation and characterization of sugilite glass from basalt for α-amylase immobilization, statistical optimization of the immobilization process and description of free and immobilized enzyme

“…The optimum temperature for α-amylase enzyme catalytic activity of both free and A-BSF enzyme was seen at 50 C (Figure 5). Similarly (Zusfahair et al, 2020), noted that there is no change in the optimum reaction temperature (40 C) before and after immobilization of bacterial amylase. By raising the temperature to 70 C, there is an obvious loss of free enzyme activity by 3.2-fold which is higher than the loss of A-BSF activity (1.4-fold).…”

Polymers as Encapsulating Agents and Delivery Vehicles of Enzymes

“…It is more difficult to extrude through the syringe needle when the viscosity is high, and, consequently, the formation of microcapsules is not uniform. Zusfahair et al [ 58 ] studied the immobilization of amylase from Aspergillus oryzae and observed a reduction in immobilization yield while increasing alginate concentration. Conformational changes in the entrapped enzyme and limitation of substrate mass transfer to the microcapsules can be caused by increasing sodium alginate concentration.…”

Immobilization Optimization and Characterization of Immobilized Lipase from Lysinibacillus macroides FS1 for Biodiesel Production

“…polymers) or inorganic support materials are used for entrapment among that sodium alginate is most widely used polymer due to its low cost, easy to use, mild gelling properties, biocompatibility, thermostability, effective particle size, availability and non toxicity (Ding et al,2020). Hence lipase entrapment in calcium alginate beads has shown to be relatively safe and straightforward techniques so in this study lipase was immobilized using calcium alginate beads (Zusfahair et al, 2020). Energy is key factor of economic development of nation.…”

“…There was an increase in immobilization efficiency as the concentration of alginate increases from 1% to 4% (w/v). The lower alginate concentration of 1% resulted in the formation of fragile beads which got disrupted easily and had larger pore sizes resulting in leakage of the enzyme from the beads thereby lowering enzyme activity (Malhotra andBasir, 2020 andZusfahair et al, 2020).The concentration calcium chloride influences the stability and porosity of the beads. Maximum immobilization efficiency and yield (69% & 43%) was recorded from beads prepared from 200mM calcium chloride.…”