Design, synthesis and characterization of enzyme-analogue-built polymer catalysts as artificial hydrolases

Figures & data

Figure 1. Discontinuate word arrangement of amino acids in peptide chain.

Figure 2. Catalytic triad and hydrophobic pocket in chymotrypsin.

Figure 3. Catalytic mechanism of chymotrypsin.

Figure 4. Reaction coordinate diagram for an enzyme-catalyzed reaction.

Figure 5. Michaelis–Menten kinetics: (a) enzyme catalyzed reaction and (b) an ordinary non-catalyzed reaction or a chemically catalyzed reaction.

Figure 6. Schematic diagram of the molecular imprinting process.

Figure 7. Some metal-mediated imprint interactions.

Table 1. Summary of analytical techniques for rational MIP assessment.

Pre-polymerization
Technique	Information
NMR spectroscopy	H-bonding [24], self-association [24], π-π stacking [25], ionic interactions [26]
UV spectroscopy	Complex events and addition of crosslinker [27]
Molecular modeling	Screening of functional monomer libraries for suitable interaction with given template [28]
Post-polymerization
Technique	Information
FT-IR spectroscopy	Analyte rebinding [29]
Solid state NMR	Analyte rebinding [30]
Microcalorimetry	Analyte rebinding strength [31]
Combinatorial chemistry	Screening of large numbers of MIPs [32]

Figure 8. Esterase MIP with the imprints of substrate analogue as chymotrypsin mimic [46].

Figure 9. Structures of different transition state analogues.

Figure 10. Hydrolysis of p-nitrophenyl acetate [47].

Figure 11. Homogeneous and heterogeneous esterase MIPs with the imprints of methyl hydrogen p-nitrobenzylphosphonate [52].

Figure 12. Heterogeneous esterase MIP [52].

Figure 13. Water soluble homogeneous esterase MIP with the imprints of phenyl-1-enzyloxycarbonylamino-3-methylpentylphosphonate [48].

Figure 14. Water soluble homogeneous alkylated and non-alkylated esterase MIPs.

Figure 15. Esterase MIP with the imprints of chiral phosphonate analogue of phenylalanine as TSA [6].

Figure 16. Esterase MIP synthesized using phenyl 1-benzyloxycarbonylamino-3-methylpentylphosphonate TSA [54].

Figure 17. Esterase MIP synthesized using N- (N-benzyloxycarbonyl-L-leucinoyl) anthranilic acid GSA [54].

Figure 18. Water soluble and water insoluble MIPs [55].

Figure 19. Water soluble esterase MIP’ with the imprints of phenyl-1-benzyloxycarbonyl-3-methylpentyl phosphonate [56].

Figure 20. Stoichiometric non-covalent imprinting strategy employing a monomer and bifunctional TSA phosphonate ester [57].

Figure 21. First reported chymotrypsin mimc [41,42].

Figure 22. Hydrophilic and hydrophobic carrier supports designed by Lee et al.

Figure 23. Imidazole-containing DVB crosslinked esterase MIP for the esterolysis of p-nitrophenyl acetate [51].

Figure 24. Polymer catalyst with TSA and the imprints of TSA [59, 60].

Figure 25. Preparation of a TSA imprinted catalyst by labile covalent binding and non-covalent binding [59,60].

Figure 26. Molecular imprinting using template monomer [58].

Figure 27. Esterolysis of long chain ester [50].

Figure 28. Introduction of nucleophilic 4-(N,N-dimethylamino)pyridines as functional monomer in imprinting [61].

Figure 29. Diels-Alder reaction using endo- and exo-TSA as template molecules.

Figure 30. Synthesis of trifunctional mimic of chymotrypsin.

Figure 31. Enantioselectivity of chymotrypsin mimic polymer catalyst.

Figure 32. Polymer catalysts with different crosslinking agents.

Figure 33. Polymer catalysts with the π-stacking interactions of pyridine.

Figure 34. Polymer catalysts in catalytic peptidolysis.

Table 2. Molecularly imprinted catalysts versus enzymes.

Sl. No.	MIPs vs enzymes
1	Specificity	Enzymes ≫> MIPs
2	Catalytic efficiency	Enzymes ≫> MIPs
3	Production and purification	Enzymes ≪< MIPs
4	Binding site homogeneity	Enzymes ≫> MIPs
5	Moderate cost and ease of preparation	Enzymes ≪< MIPs
6	Handling and stability	Enzymes ≪< MIPs
7	Regeneration and reusability	Enzymes ≪< MIPs
8	Storage and Shelf-life	Enzymes ≪< MIPs

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