Flow-Through Macroporous Polymer Monoliths Containing Artificial Catalytic Centers Mimicking Chymotrypsin Active Site

Abstract

Synthetic catalysts that could compete with enzymes in term of the catalytic efficiency but surpass them in stability have a great potential for the practical application. In this work, we have developed a novel kind of organic catalysts based on flow-through macroporous polymer monoliths containing catalytic centers that mimic the catalytic site of natural enzyme chymotrypsin. It is known that chymotrypsin catalytic center consists of L-serine, L-histidine, and L-aspartic acid and has specificity to C-terminal residues of hydrophobic amino acids (L-phenylalanine, L-tyrosine, and L-tryptophan). In this paper, we have prepared the macroporous polymer monoliths bearing grafted polymer layer on their surface. The last one was synthesized via copolymerization of N-methacryloyl-L-serine, N-methacryloyl-L-histidine, and N-methacryloyl-L-aspartic acid. The spatial orientation of amino acids in the polymer layer, generated on the surface of monolithic framework, was achieved by coordinating amino acid-polymerizable derivatives with cobalt (II) ions without substrate-mimicking template and with its use. The conditions for the preparation of mimic materials were optimized to achieve a mechanically stable system. Catalytic properties of the developed systems were evaluated towards the hydrolysis of ester bond in a low molecular substrate and compared to the results of using chymotrypsin immobilized on the surface of a similar monolithic framework. The effect of flow rate increase and temperature elevation on the hydrolysis efficiency were evaluated for both mimic monolith and column with immobilized enzyme.

Molecular Imprinting on Nanozymes for Sensing Applications

“…Imprinted polymers have been combined with amino acids, peptides, metals, among others, in synergic strategies to realize the potential of artificial enzymes [ 69 , 70 , 71 ]. For example, peroxidase mimics are useful in the detection of the relevant biomarker for tumor diagnostic 5-hydroxyindole-3-acetic acid (5-HIAA), which is an indoleamine metabolite and is simultaneously a substrate for peroxidase activity [ 72 ].…”

Macroporous Polymer Monoliths in Thin Layer Format

“…At present, existing flow-through monoliths of various geometries (discs, rods, tubes, and capillary columns) with axial or radial flow have been developed for analytical and industrial applications [23][24][25][26]. A huge number of modern publications on the application of macroporous monoliths are devoted to the various flow-through techniques such as chromatographic separation of small molecules [27,28], synthetic polymers [29], and biological objects (proteins [30], peptides [31], oligosaccharides [32], DNA [33], viruses [34]), solid-phase extraction of wide range of compounds [35][36][37], and flow-through biocatalysis [38][39][40].…”

Immobilization of trypsin onto porous methacrylate-based monolith for flow-through protein digestion and its potential application to chiral separation using liquid chromatography

“…Besides CSP, a large number of trypsin-immobilized monoliths were also reported for protein digestion and peptide analysis [ 2 , 20 , 21 , 22 ]. In addition to trypsin, immobilization of chymotrypsin on monoliths or hybrid magnetic materials is also used for protein hydrolyzation [ 23 , 24 ], and peptide synthesis [ 25 ], but is rarely applied to CSP [ 26 ]. This work focuses on examining the potential of trypsin-immobilized onto the methacrylate-based organic polymer monolithic columns as a nanobiocatalyst microreactor for high-speed and online digestion of proteins, and chiral stationary phase (CSP) for the separation of natural products.…”