A NEW APPROACH TO EVALUATE IMMOBILIZATION OF β-GALACTOSIDASE ON EUPERGIT^®C: STRUCTURAL, KINETIC, AND THERMAL CHARACTERIZATION

Abstract

This study aimed to evaluate β-galactosidase immobilization. For this purpose, the ionic strength of the buffer, reaction time, amount of the immobilization support, and pH were evaluated by a central composite design. Assay 8, which consisted of 1.5 mol L −1 phosphate buffer (pH 7.5) and a reaction time of 2 h, produced the maximum yield. Eupergit ® C (400 mg) was subsequently used as an immobilization support. Immobilization kinetics wereinvestigated, and a significant increase in the yield was obtained after immobilization compared with that obtained from assay 8 (22.0 U mL −1 vs. 15.6 U mL −1). The enzyme efficiency of actuation was evaluated using o-nitrophenyl-β-d-galactopyranoside and lactose, with lactose providing better results. The reuse of β-galactosidase was evaluated, and more than 50% of the initial enzyme activity was maintained after five cycles of use. Enzyme characterization revealed that immobilization improved some aspects of the thermostability of β-galactosidase.

Immobilization of Glucose Oxidase on Eupergit C: Impact of Aeration, Kinetic and Operational Stability Studies of Free and Immobilized Enzyme

“…Original scientific paper Received: May 29, 2018 Accepted: December 5, 2018 stable and possesses good chemical and mechanical properties, such as simple immobilization procedure and high binding capacity 17 . Eupergit C is a naturally porous polymer 18 and immobilization on it can be achieved without the need for additional reagents by forming strong covalent linkages between Eupergit epoxy groups and enzyme nucleophilic groups 19 .…”

Biological Approaches for Extraction of Bioactive Compounds From Agro-industrial By-products: A Review

“…To further reduce the cost of applying processes using enzymes, agro-industrial waste available in large quantities can be used in the production of enzymes (Lemes et al, 2016a) using simpler purification protocols or coupling techniques to purify the target product (Lemes et al, 2014;Lemes et al, 2019). Another factor that supports the application of enzymes and cost reduction in the process is the possibility of their immobilization, resulting in their recyclable use, allowing their application in defined cycles, and maintaining their selectivity, catalytic activity, and the generation of products in large quantities (Braga et al, 2014;Basso and Serban, 2019).…”

Enzyme immobilization: polymer–solvent–enzyme compatibility

“…[130][131][132] Synthetic polymers often require surface modification to achieve post immobilization. 133 Some commonly used synthetic polymers are PVA, 134 polyvinyl acetate (PVAc), 81 polyimide (PI), 91 polyacrylates (e.g., poly(methyl methacrylate) (PMMA), Eupergit ® -C), 135,136 polyamide (e.g. nylon 6), 137 polyacrylonitrile (PAN), 138 polyaniline (PANi), 139 poly(ethylene glycol) (PEG), 140 and poly(lactic-co-glycolic acid) (PLGA).…”