Amidase activity of phosphonate analogue imprinted chymotrypsin mimics in shape-selective, substrate-specific and enantioselective amidolysis of l -phenylalanine- p -nitroanilides

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

“…The amidase MIP did not exhibit considerable loss in enzymatic activity in amidolysis even after three years of shelf life. The mimic showed only 6% decrease after three years of shelf-life [63].…”

“…N. Kirsch et al in 2009, reported product analogue imprinted polymer catalyzed Diels-Alder cycloaddition reaction [63] reaction of 1, 3-butadiene carbamic acid benzyl ester and N,N-dimethylacrylamide ( Figure 29). Recognized transition state analogues for the endo-and exo-reaction pathways were used as templates for the synthesis of molecularly imprinted methacrylic acid-divinylbenzene copolymers.…”

“…In our lab, we carried out the hydrolysis of the amide bonds in amino acid p-nitroanilides and dipeptides using MIPs. We synthesized EGDMA crosslinked chymotrypsin mimic polymer catalysts by incorporating the imprints of stable phosphonate TSA, phenyl-1-benzyloxycarbonylaminoethylphosphonate and the catalytic triad-His, Ser, Asp-using methacrylated amino acid monomers ( Figure 30) [63]. The individual and combined effects of imidazole, carboxyl and hydroxyl groups in the polymer matrix on the catalytic amidolysis of chymotrypsin specific and non-specific amino acid p-nitroanilides were investigated in detail by synthesizing mono, bi and trifunctional polymer catalysts.…”

“…The functional groups MIPsthe highly appealing alternativescan easily be regenerated without loss of affinity, and hence are highly useful for continuous use. The reusability of the spent amidase MIP was investigated by our group by carrying out the amidolytic reaction after regenerating the polymer [63]. The rate acceleration of the reaction was evaluated for each cycle.…”

“…Natural chymotrypsin is denatured in the temperature range around 50 C. But it was found that, the mimics were stable up to 130 C and at higher temperatures they showed diminished amidase activity [63].…”