An Enzyme Model Which Mimics Chymotrypsin and N-Terminal Hydrolases

Abstract

Enzymes are the most efficient and specific catalysts to date. Although they have been thoroughly studied for years, building a true enzyme mimic remains a challenging and necessary task. Here, we show how a three-dimensional geometry analysis of the key catalytic residues in natural hydrolases has been exploited to design and synthesize small-molecule artificial enzymes which mimic the active centers of chymotrypsin and N-terminal hydrolases. The optimized prototype catalyzes the methanolysis of the acyl enzyme mimic with a half-life of only 3.7 min at 20 °C, and it is also able to perform the transesterification of vinyl acetate at room temperature. DFT studies and X-ray diffraction analysis of the catalyst bound to a transition state analogue proves the similarity with the geometry of natural hydrolases.

Selective Hydrolysis of Aryl Esters under Acidic and Neutral Conditions by a Synthetic Aspartic Protease Mimic

“…Synthetic esterases are one of the most studied enzyme-mimicking cataysts. , Recent examples, built on peptide or purely abiotic scaffolds, commonly contain either a Lewis acidic metal ion such as zinc to activate water ,− or a catalytic triad to mimic those found in serine proteases. − Although these studies used different reaction conditions, the catalytic efficiency of MINP­( 1 + 5e ) ( k cat / K m = 343 M –1 s –1 at pH 7, 25 °C) compared favorably with those of previously reported systems for PNPA hydrolysis ( k cat / K m = 2–100 M –1 s –1 at pH 6.2–8.0, 22–25 °C), and was close to that (450 M –1 s –1 ) of a synthetic esterase (formed from amyloid fibrils) at pH 8, 38 °C, under high pressure (200 MPa) . The catalytic turnover number (TON) of MINP­( 1 + 5e ) was >510 (Figure b), also significantly higher than those in the literature (mostly >10–20). − …”

Engineering a Feruloyl–Coenzyme A Synthase for Bioconversion of Phenylpropanoid Acids into High-Value Aromatic Aldehydes

“…The acrylic group is acylated with CoA to form an acyl−CoA complex under the FCS enzyme fixation in the ligase domain. 53,54 Precisely locating the substrate-binding domain in this study provided the basis for subsequent FCS enzyme engineering.…”

“…From a structural perspective, the catalytic mechanism of FCS could be described as CoA and substrate bound with the CoA-binding and substrate-binding domains, respectively. The acrylic group is acylated with CoA to form an acyl–CoA complex under the FCS enzyme fixation in the ligase domain. , Precisely locating the substrate-binding domain in this study provided the basis for subsequent FCS enzyme engineering.…”

Transesterification of Non‐Activated Esters Promoted by Small Molecules Mimicking the Active Site of Hydrolases

“…For this first step to take place efficiently, we think that the association of the carbonyl group with the oxyanion hole is critical. First‐generation catalyst 1 had already shown a low association constant for ethyl acetate (0.79 M −1 ) [10] . However, attempts to measure the association constant between catalyst 10 and ethyl acetate by 1 H NMR titration failed.…”