Kaiser, E.T.

Biopolymers 1990, 29, 39-43, 10.1002/bip.360290107

The semisyntehtic enzyme 6 was prepared by alkylation of the cysteine‐25 sulfhydryl group of papain with the bipyridine 5 and was shown to stoichiometrically bind copper ion; 7 catalyzed the autoxidation of ascorbic acid derivatives with saturation kinetics approximately 20‐fold faster than a model system using 3‐Cu(II).

Hromadová, M.

J. Electroanal. Chem. 2020, 859, 113882, 10.1016/j.jelechem.2020.113882

Electrochemical properties were studied for [(η6-arene)Ru(1,10-phenanthroline)Cl]Cl (arene = C6H5(CH2)2NHCOCH2Cl) organometallic complex 1, protein Papain PAP and its conjugate with organometallic complex 1-PAP. The latter can serve as an artificial metalloenzyme with catalytic activity in transfer hydrogenation. This work demonstrates that AC voltammetry and electrochemical impedance spectroscopy can be used as fast tools to screen the catalytic ability of 1-PAP electrochemically by studies of the catalytic hydrogen evolution reaction (HER). Proteins are known to catalyze this process, but we have shown that additional HER signal associated with the catalytic activity of 1 is observed for its conjugate with Papain 1-PAP.

Kaiser, E.T.

J. Am. Chem. Soc. 1987, 109, 606-607, 10.1021/ja00236a062

n/a

Kaiser, E.T.; Sasaki, T.

J. Am. Chem. Soc. 1989, 111, 380-381, 10.1021/ja00183a065

n/a

Kazlauskas, R.J.

Chem. - Eur. J. 2006, 12, 1587-1596, 10.1002/chem.200501413

Carbonic anhydrase is a zinc metalloenzyme that catalyzes the hydration of carbon dioxide to bicarbonate. Replacing the active‐site zinc with manganese yielded manganese‐substituted carbonic anhydrase (CA[Mn]), which shows peroxidase activity with a bicarbonate‐dependent mechanism. In the presence of bicarbonate and hydrogen peroxide, (CA[Mn]) catalyzed the efficient oxidation of o‐dianisidine with kcat/KM=1.4×106 m−1 s−1, which is comparable to that for horseradish peroxidase, kcat/KM=57×106 m−1 s−1. CA[Mn] also catalyzed the moderately enantioselective epoxidation of olefins to epoxides (E=5 for p‐chlorostyrene) in the presence of an amino‐alcohol buffer, such as N,N‐bis(2‐hydroxyethyl)‐2‐aminoethanesulfonic acid (BES). This enantioselectivity is similar to that for natural heme‐based peroxidases, but has the advantage that CA[Mn] avoids the formation of aldehyde side products. CA[Mn] degrades during the epoxidation limiting the yield of the epoxidations to <12 %. Replacement of active‐site residues Asn62, His64, Asn67, Gln92, or Thr200 with alanine by site‐directed mutagenesis decreased the enantioselectivity demonstrating that the active site controls the enantioselectivity of the epoxidation.

Kazlauskas, R.J.

Top. Organomet. Chem. 2009, 10.1007/3418_2008_1

Carbonic anhydrase binds a zinc ion in a hydrophobic active site using the imidazole groups of three histidine residues. The natural role of carbonic anhydrase is to catalyze the reversible hydration of carbon dioxide to bicarbonate, but it also catalyzes hydrolysis of esters with moderate enantioselectivity. Replacing the active-site zinc with manganese yielded manganese-substituted carbonic anhydrase (CA[Mn]), which shows peroxidase activity with a bicarbonate-dependent mechanism. In the presence of bicarbonate and hydrogen peroxide, CA[Mn] catalyzed the efficient oxidation of o-dianisidine with k cat /K M = 1.4 × 106 M−1s−1, which is comparable to that for horseradish peroxidase, k cat /K M = 57 × 106 M−1s−1. CA[Mn] also catalyzed the moderately enantioselective epoxidation of olefins to epoxides (E = 5 for p-chlorostyrene). This enantioselectivity is similar to that for natural heme-based peroxidases, but has the advantage that CA[Mn] avoids formation of aldehyde side products. CA[Mn] degrades during the epoxidation, limiting the yield of the epoxidations to <12%. Replacement of active-site residues Asn62, His64, Asn67, Gln92, or Thr200 with alanine by site-directed mutagenesis decreased the enantioselectivity showing that the active site controls enantioselectivity of the epoxidation.

Kazlauskas, R.J.

ChemCatChem 2010, 2, 953-957, 10.1002/cctc.201000159

CA confidential: Replacing the active‐site zinc in carbonic anhydrase (CA) by rhodium forms a new enzymatic catalyst for cofactor‐free hydroformylation of styrene with syn gas. Unlike free rhodium, this rhodium–protein hybrid, [Rh]–CA, is regioselective (8.4:1) for linear over branched aldehyde product, which is a 40‐fold change in regioselectivity compared to free rhodium.

Kazlauskas, R.J.

Chem. - Eur. J. 2009, 15, 1370-1376, 10.1002/chem.200801673

One useful synthetic reaction missing from nature's toolbox is the direct hydrogenation of substrates using hydrogen. Instead nature uses cofactors like NADH to reduce organic substrates, which adds complexity and cost to these reductions. To create an enzyme that can directly reduce organic substrates with hydrogen, researchers have combined metal hydrogenation catalysts with proteins. One approach is an indirect link where a ligand is linked to a protein and the metal binds to the ligand. Another approach is direct linking of the metal to protein, but nonspecific binding of the metal limits this approach. Herein, we report a direct hydrogenation of olefins catalyzed by rhodium(I) bound to carbonic anhydrase (CA‐[Rh]). We minimized nonspecific binding of rhodium by replacing histidine residues on the protein surface using site‐directed mutagenesis or by chemically modifying the histidine residues. Hydrogenation catalyzed by CA‐[Rh] is slightly slower than for uncomplexed rhodium(I), but the protein environment induces stereoselectivity favoring cis‐ over trans‐stilbene by about 20:1. This enzyme is the first cofactor‐independent reductase that reduces organic molecules using hydrogen. This catalyst is a good starting point to create variants with tailored reactivity and selectivity. This strategy to insert transition metals in the active site of metalloenzymes opens opportunities to a wider range of enzyme‐catalyzed reactions.

Kaiser, E.T.

J. Chem. Soc., Chem. Commun. 1976, 830, 10.1039/C39760000830

Copper(II) carboxypeptidase A catalyses the oxidation of ascorbic acid and this reaction is inhibited by α-benzylsuccinate, a known inhibitor of the thiolesterase action of the copper enzyme; the pH dependencies of kcat and kcat/Km are similar near pH 7 to those seen for the peptidase and esterase activities of native carboxypeptidase A.