15 publications

15 publications

An Enantioselective Artificial Metallo-Hydratase

Roelfes, G.

Chem. Sci., 2013, 10.1039/c3sc51449h

Direct addition of water to alkenes to generate important chiral alcohols as key motif in a variety of natural products still remains a challenge in organic chemistry. Here, we report the first enantioselective artificial metallo-hydratase, based on the transcription factor LmrR, which catalyses the conjugate addition of water to generate chiral β-hydroxy ketones with enantioselectivities up to 84% ee. A mutagenesis study revealed that an aspartic acid and a phenylalanine located in the active site play a key role in achieving efficient catalysis and high enantioselectivities.


Metal: Cu
Ligand type: Phenanthroline
Host protein: LmrR
Anchoring strategy: Covalent
Optimization: Genetic
Max TON: 30
ee: 84
PDB: 3F8B
Notes: ---

An NAD(P)H-Dependent Artificial Transfer Hydrogenase for Multienzymatic Cascades

Ward, T. R.

J. Am. Chem. Soc., 2016, 10.1021/jacs.6b02470


Metal: Ir
Ligand type: Cp*; Phenanthroline
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Max TON: >999
ee: >99
PDB: ---
Notes: ---

Artificial Copper Enzymes for Asymmetric Diels–AlderReactions

Kamer, P. C. J.; Laan, W.

ChemCatChem, 2012, 10.1002/cctc.201200671


Metal: Cu
Anchoring strategy: Covalent
Optimization: Chemical & genetic
Max TON: 9.6
ee: 25
PDB: 1IKT
Notes: ---

Artificial Metalloenzymes based on Protein Cavities: Exploring the Effect of Altering the Metal Ligand Attachment Position by Site Directed Mutagenesis

Distefano, M. D.

Bioorg. Med. Chem. Lett., 1999, 10.1016/S0960-894X(98)00684-2


Metal: Cu
Ligand type: Phenanthroline
Anchoring strategy: Covalent
Optimization: Genetic
Max TON: 1 to 4
ee: 61 to 94
PDB: ---
Notes: Varied attachment position

Artificial Metalloenzymes with the Neocarzinostatin Scaffold: Toward a Biocatalyst for the Diels–Alder Reaction

Mahy, J.-P.; Ricoux, R.

ChemBioChem, 2016, 10.1002/cbic.201500445


Metal: Cu
Ligand type: Phenanthroline
Anchoring strategy: Supramolecular
Optimization: ---
Max TON: 33
ee: ---
PDB: ---
Notes: Up to endo/exo ratio 62:38

A Semisynthetic Metalloenzyme based on a Protein Cavity that Catalyzes the Enantioselective Hydrolysis of Ester and Amide Substrates

Distefano, M. D.

J. Am. Chem. Soc., 1997, 10.1021/JA970820K

In an effort to prepare selective and efficient catalysts for ester and amide hydrolysis, we are designing systems that position a coordinated metal ion within a defined protein cavity. Here, the preparation of a protein-1,10-phenanthroline conjugate and the hydrolytic chemistry catalyzed by this construct are described. Iodoacetamido-1,10-phenanthroline was used to modify a unique cysteine residue in ALBP (adipocyte lipid binding protein) to produce the conjugate ALBP-Phen. The resulting material was characterized by electrospray mass spectrometry, UV/vis and fluorescence spectroscopy, gel filtration chromatography, and thiol titration. The stability of ALBP-Phen was evaluated by guanidine hydrochloride denaturation experiments, and the ability of the conjugate to bind Cu(II) was demonstrated by fluorescence spectroscopy. ALBP-Phen-Cu(II) catalyzes the enantioselective hydrolysis of several unactivated amino acid esters under mild conditions (pH 6.1, 25 °C) at rates 32−280-fold above the background rate in buffered aqueous solution. In 24 h incubations 0.70 to 7.6 turnovers were observed with enantiomeric excesses ranging from 31% ee to 86% ee. ALBP-Phen-Cu(II) also promotes the hydrolysis of an aryl amide substrate under more vigorous conditions (pH 6.1, 37 °C) at a rate 1.6 × 104-fold above the background rate. The kinetics of this amide hydrolysis reaction fit the Michaelis−Menten relationship characteristic of enzymatic processes. The rate enhancements for ester and amide hydrolysis reported here are 102−103 lower than those observed for free Cu(II) but comparable to those previously reported for Cu(II) complexes.


Metal: Cu
Ligand type: Phenanthroline
Anchoring strategy: Covalent
Optimization: ---
Max TON: 1 to 8
ee: 39 to 86
PDB: ---
Notes: ---

Chemical Conversion of a DNA-Binding Protein into a Site-Specific Nuclease

Sigman, D. S.

Science, 1987, 10.1126/science.2820056


Metal: Cu
Ligand type: Phenanthroline
Anchoring strategy: Covalent
Optimization: ---
Reaction: Oxidative cleavage
Max TON: <1
ee: ---
PDB: ---
Notes: Engineered sequence specificity

Conversion of a Helix-Turn-Helix Motif Sequence-Specific DNA Binding Protein into a Site-Specific DNA Cleavage Agent

Ebright, R. H.; Gunasekeram, A.

Proc. Natl. Acad. Sci. U. S. A., 1990, 10.1073/pnas.87.8.2882


Metal: Cu
Ligand type: Phenanthroline
Anchoring strategy: Covalent
Optimization: ---
Reaction: Oxidative cleavage
Max TON: <1
ee: ---
PDB: ---
Notes: Engineered sequence specificity

Cross-Regulation of an Artificial Metalloenzyme

Ward, T. R.

Angew. Chem., Int. Ed., 2017, 10.1002/anie.201702181


Metal: Ir
Ligand type: Cp*; Phenanthroline
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Max TON: 96
ee: ---
PDB: ---
Notes: Cross-regulated reduction of the antibiotic enrofloxacin by an ArM.

Enantioselective Artificial Metalloenzymes by Creation of a Novel Active Site at the Protein Dimer Interface

Roelfes, G.

Angew. Chem., Int. Ed., 2012, 10.1002/anie.201202070


Metal: Cu
Ligand type: Bipyridine; Phenanthroline
Host protein: LmrR
Anchoring strategy: Covalent
Optimization: Genetic
Max TON: 32.7
ee: 97
PDB: 3F8B
Notes: ---

Piano-Stool d(6)-Rhodium(III) Complexes of Chelating Pyridine-Based Ligands and their Papain Bioconjugates for the Catalysis of Transfer Hydrogenation of Aryl Ketones in Aqueous Medium

Mangiatordi, G. F.; Salmain, M.

J. Mol. Catal. B: Enzym., 2015, 10.1016/j.molcatb.2015.10.007


Metal: Rh
Ligand type: Cp*; Phenanthroline
Host protein: Papain (PAP)
Anchoring strategy: Covalent
Optimization: Chemical
Max TON: 30
ee: 9
PDB: ---
Notes: ---

Metal: Rh
Ligand type: Cp*; Di(2-pyridyl)
Host protein: Papain (PAP)
Anchoring strategy: Covalent
Optimization: Chemical
Max TON: 20
ee: 5
PDB: ---
Notes: ---

Receptor-Based Artificial Metalloenzymes on Living Human Cells

Ghattas, W.; Mahy, J.-P.

J. Am. Chem. Soc., 2018, 10.1021/jacs.8b04326


Metal: Cu
Ligand type: Phenanthroline
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Max TON: 24
ee: 35
PDB: ---
Notes: ---

Supramolecular Assembly of Artificial Metalloenzymes Based on the Dimeric Protein LmrR as Promiscuous Scaffold

Roelfes, G.

J. Am. Chem. Soc., 2015, 10.1021/jacs.5b05790


Metal: Cu
Ligand type: Phenanthroline
Host protein: LmrR
Anchoring strategy: Supramolecular
Optimization: Genetic
Max TON: 11.1
ee: 94
PDB: 3F8B
Notes: ---

Synthesis of a Heterogeneous Artificial Metallolipase with Chimeric Catalytic Activity

Filice, M.

Chem. Commun., 2015, 10.1039/C5CC02450A


Metal: Cu
Ligand type: Phenanthroline
Anchoring strategy: Covalent
Optimization: Genetic
Max TON: 411
ee: 92
PDB: ---
Notes: ArM is immobilized on Sepabeads. Endo/exo = 93.5%

Metal: Cu
Ligand type: Phenanthroline
Anchoring strategy: Covalent
Optimization: Genetic
Reaction: Reduction
Max TON: ---
ee: ---
PDB: ---
Notes: Cascade reaction: Ester hydrolysis (natural function of the host protein) followed by reduction (function of the designed ArM).

(η6-Arene) Ruthenium(II) Complexes and Metallo-Papain Hybrid as Lewis Acid Catalysts of Diels–Alder Reaction in Water

Salmain, M.

Dalton Trans., 2010, 10.1039/c001630f

Covalent embedding of a (η6-arene) ruthenium(II) complex into the protein papain gives rise to a metalloenzyme displaying a catalytic efficiency for a Lewis acid-mediated catalysed Diels–Alder reaction enhanced by two orders of magnitude in water.


Metal: Ru
Ligand type: Benzene; Phenanthroline
Host protein: Papain (PAP)
Anchoring strategy: Covalent
Optimization: Chemical
Max TON: 440
ee: ---
PDB: ---
Notes: TOF = 220 h-1