Immobilized Alcalase on Micron- and Submicron-Sized Alginate Beads as a Potential Biocatalyst for Hydrolysis of Food Proteins

Abstract

Enzymatic hydrolysis of food proteins is convenient method to improve their functional properties and physiological activity. Herein, the successful covalent attachment of alcalase on alginate micron and submicron beads using the carbodiimide based chemistry reaction and the subsequent application of the beads for egg white and soy proteins hydrolysis were studied. In addition to the electrostatic extrusion technique (EE) previously used by others, the potential utilization of a novel ultrasonic spray atomization technique without drying (UA) and with drying (UAD) for alginate submicron beads production has been attempted. The immobilization parameters were optimized on microbeads obtained by EE technique (803 ± 23 µm) with respect to enzyme loading and alcalase activity. UA and UAD techniques resulted in much smaller particles (607 ± 103 nm and 394 ± 51 nm in diameter, respectively), enabling even higher enzyme loading of 671.6 ± 4 mg g−1 on the carrier and the highest immobilized alcalase activity of 2716.1 IU g−1 in the standard reaction. The UAD biocatalyst exhibited also better performances in the real food system based on egg white or soy proteins. It has been shown that the immobilized alcalase can be reused in seven successive soy protein hydrolysis cycles with a little decrease in the activity.

Biomass-derived functional materials as carriers for enzymes: towards sustainable and robust biocatalysts

“…217,218 An attractive approach to increase the stability of enzymatic processes and economic feasibility in terms of reusability can be achieved with the use of alginate-based supports for enzyme immobilization. [219][220][221][222][223][224][225][226] The mechanical characteristics and robustness of alginate-based supports can be enhanced further using polymer blending (much like cellulose and chitosan). These alginate forms have been demonstrated to increase thermal stability, enzyme activity, and reusability.…”

Immobilization of Horseradish Peroxidase on Magnetite-Alginate Beads to Enable Effective Strong Binding and Enzyme Recycling during Anthraquinone Dyes’ Degradation

“…For the immobilization of HRP on MABs the optimal conditions from our previous work have been used [40]. To activate 0.5 g of MABs, they were treated with 10 mg of EDAC in 10 mL of 50 mM Tris-HCl buffer pH 8.5 for 30 min under gentle stirring (150 rpm) at 25 • C. After reaction of the MABs with EDAC, further reactivity of EDAC with carboxyl groups was quenched by addition of 2-mercaptoethanol (10-fold in excess of EDAC) for 10 min.…”

“…Herein, we propose embedding Mag into hydrogel mm-or µm-scale particles (beads) which combine advantages of both (gel particles and Mag)-rapid separation by external magnetic field and/or gravitational settling due to high density, stability against dissolution especially at lower pH of solution, amplified enzyme activity and a number of available methods able to give large amounts of beads under mild and simple preparation conditions. Of our particular interest is entrapment of Mag in calcium alginate beads, since covalent binding of enzymes to calcium alginate using carbodiimide chemistry has been described as a suitable method to achieve stabilization and good performance of several enzymes including alcalase [40], β-galactosidase [22], acetylcholinesterase [23], α-amylase [24] and others. According to our knowledge, this is the first time that peroxidase is covalently immobilized on Mag-alginate beads.…”

Palladated Cyclodextrin Nanosponge-Alginate Dual Bead as an Efficient Catalyst for Hydrogenation of Nitroarenes in Aqueous Solution

“…While sodium alginate hydrogels/beads find significant utility in cell encapsulation and drug delivery, they have also gained prominence in catalysis [ 8 , 33 ] due to their hydrophilicity, affordability, and relative durability. Additionally, various composite hydrogels and beads based on Alg have been reported, serving as effective matrices for immobilizing or encapsulating catalytic species [ 33 , 34 , 35 ].…”