Abstract
Proteases are involved in several crucial biological processes and reported to have important physiological functions. They also have multifarious applications in different industries. The immobilized form of the enzyme further improves its industrial applicability. Here, we report covalent immobilization of a novel cysteine endopeptidase (procerain B) on amberlite MB-150 beads through glutaraldehyde by Schiff base linkage. The immobilized product was examined extensively by Fourier Transform Infrared Spectroscopy (FTIR), Scanning electron microscopy (SEM) and Energy Dispersive X-ray (EDX) analysis. The characterization of the immobilized product showed broader pH and thermal optima compared to the soluble form of the enzyme. The immobilized form of procerain B also showed lower Km (180.27±6 µM) compared to the soluble enzyme using azocasein as substrate. Further, immobilized procerain B retains 38.6% activity till the 10th use, which strongly represents its industrial candidature.
Highlights
Enzyme immobilization is a process of physical localization of enzymes to a defined surface which helps to improve several enzymatic properties and enhance their operational performance without disturbing their catalytic activity [1,2,3,4,5,6]
We have reported purification and characterized a novel cysteine endopeptidase from the latex of a medicinal plant Calotropis procera [21]
The amberlite MB-150 beads, azocasine, glutaraldehyde, protease inhibitor, trichloroacetic acid (TCA), Sodium tetrathionate (STT) and Bradford reagent were purchased from Sigma Chemicals Co
Summary
Enzyme immobilization is a process of physical localization of enzymes to a defined surface which helps to improve several enzymatic properties and enhance their operational performance without disturbing their catalytic activity [1,2,3,4,5,6]. Immobilization of enzyme allow recovery and reusability of the enzyme making the overall process controllable and economical [7,8]. Proteases have several applications in food, dairy and detergent industries. It has a wide range of applicability in medicines, energy production and environmental control [9,10,11]. Proteases are one of the most common industrial enzymes which have wide range of applications varying from food to pharma industries [12,13,14,15]. Some neutral and alkaline proteases are used for the recovery of meat from butchering which are used in several canned soups [17,18]. Certain proteases are used as ingredients of chocolates, cakes and some canned drinks to enhance the flavor
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