46 publications

46 publications

8-Amino-5,6,7,8-tetrahydroquinoline in Iridium(III) Biotinylated Cp* Complex as Artificial Imine Reductase

Rimoldi, I.

New J. Chem., 2018, 10.1039/C8NJ04558E

Diamine ligands I–IV coordinated to an iridium metal complex with the biotin moiety anchored to the Cp* ring were investigated. This strategy, in contrast to the traditional biotin–streptavidin technology that uses a biotinylated ligand in the artificial imine reductase, is practical for envisaging how the enantiodiscrimination by different Streptavidin (Sav) mutants could influence the chiral environment of the metal cofactor. Only in the case of (R)-CAMPY IV did the chirality at the metal centre and the second coordination sphere environment, which was dictated by the host protein, operate in a synergistic way, producing better enantioselectivity at a S112M Sav catalyst/catalyst ratio of 1.0 : 2.5. Under these optimized conditions, the artificial imine reductase afforded a good enantiomeric excess (83%) in the asymmetric transfer hydrogenation of 6,7-dimethoxy-1-methyl-3,4-dihydroisoquinoline.


Metal: Ir
Ligand type: Cp*; Diamine
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Max TON: 32
ee: 83
PDB: ---
Notes: ---

Metal: Ir
Ligand type: Cp*; Diamine
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Max TON: 99
ee: 13
PDB: ---
Notes: ---

A Cell-Penetrating Artificial Metalloenzyme Regulates a Gene Switch in a Designer Mammalian Cell

Fussenegger, M.; Matile, S.; Ward, T. R.

Nat. Commun., 2018, 10.1038/s41467-018-04440-0

Complementing enzymes in their native environment with either homogeneous or heterogeneous catalysts is challenging due to the sea of functionalities present within a cell. To supplement these efforts, artificial metalloenzymes are drawing attention as they combine attractive features of both homogeneous catalysts and enzymes. Herein we show that such hybrid catalysts consisting of a metal cofactor, a cell-penetrating module, and a protein scaffold are taken up into HEK-293T cells where they catalyze the uncaging of a hormone. This bioorthogonal reaction causes the upregulation of a gene circuit, which in turn leads to the expression of a nanoluc-luciferase. Relying on the biotin–streptavidin technology, variation of the biotinylated ruthenium complex: the biotinylated cell-penetrating poly(disulfide) ratio can be combined with point mutations on streptavidin to optimize the catalytic uncaging of an allyl-carbamate-protected thyroid hormone triiodothyronine. These results demonstrate that artificial metalloenzymes offer highly modular tools to perform bioorthogonal catalysis in live HEK cells.


Metal: Ru
Ligand type: Cp; Quinoline
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Genetic
Reaction: Deallylation
Max TON: 33
ee: ---
PDB: ---
Notes: ---

A Designed Heme-[4Fe-4S] Metalloenzyme Catalyzes Sulfite Reduction like the Native Enzyme

Lu, Y.

Science, 2018, 10.1126/science.aat8474

Multielectron redox reactions often require multicofactor metalloenzymes to facilitate coupled electron and proton movement, but it is challenging to design artificial enzymes to catalyze these important reactions, owing to their structural and functional complexity. We report a designed heteronuclear heme-[4Fe-4S] cofactor in cytochrome c peroxidase as a structural and functional model of the enzyme sulfite reductase. The initial model exhibits spectroscopic and ligand-binding properties of the native enzyme, and sulfite reduction activity was improved—through rational tuning of the secondary sphere interactions around the [4Fe-4S] and the substrate-binding sites—to be close to that of the native enzyme. By offering insight into the requirements for a demanding six-electron, seven-proton reaction that has so far eluded synthetic catalysts, this study provides strategies for designing highly functional multicofactor artificial enzymes.


Metal: Fe
Host protein: Cytochrome c peroxidase
Anchoring strategy: Dative
Optimization: Chemical & genetic
Reaction: Sulfite reduction
Max TON: ---
ee: ---
PDB: ---
Notes: Designed heteronuclear heme-[4Fe-4S] cofactor in cytochrome c peroxidase

An Artificial Heme Enzyme for Cyclopropanation Reactions

Roelfes, G.

Angew. Chem., Int. Ed., 2018, 10.1002/anie.201802946


Metal: Fe
Ligand type: Protoporphyrin IX
Host protein: LmrR
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Reaction: Cyclopropanation
Max TON: 449
ee: 51
PDB: 6FUU
Notes: ---

An Artificial Metalloenzyme for Carbene Transfer Based on a Biotinylated Dirhodium Anchored Within Streptavidin

Ward, T. R.

Cat. Sci. Technol., 2018, 10.1039/C8CY00646F


Metal: Rh
Ligand type: Carboxylate
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Reaction: Cyclopropanation
Max TON: ~60
ee: ---
PDB: ---
Notes: Cyclopropanation reaction was also performed in the E. coli periplasm.

Metal: Rh
Ligand type: Carboxylate
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Reaction: C-H insertion
Max TON: ~60
ee: ---
PDB: ---
Notes: ---

A Noncanonical Proximal Heme Ligand Affords an Efficient Peroxidase in a Globin Fold

Green, A. P.; Hilvert, D.

J. Am. Chem. Soc., 2018, 10.1021/jacs.7b12621


Metal: Fe
Host protein: Myoglobin (Mb)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Reaction: Oxidation
Max TON: ~1650
ee: ---
PDB: 5OJ9
Notes: Oxidation of amplex red

Artificial Heme Enzymes for the Construction of Gold-Based Biomaterials

Lombardi, A.; Nastri, F.

Int. J. Mol. Sci., 2018, 10.3390/ijms19102896


Metal: Fe
Ligand type: Amino acid; Porphyrin
Anchoring strategy: Covalent
Optimization: Chemical & genetic
Reaction: Oxidation
Max TON: ---
ee: ---
PDB: ---
Notes: Immobilization of the ArM on gold surfaces via a lipoic acid anchor.

Artificial Metalloenzyme Design with Unnatural Amino Acids and Non-Native Cofactors

Review

Wang, J.

ACS Catal., 2018, 10.1021/acscatal.7b03754


Notes: ---

Artificial Metalloenzymes as Catalysts for Oxidative Lignin Degradation

Jarvis, A. G.

ACS Sustainable Chem. Eng., 2018, 10.1021/acssuschemeng.8b03568


Metal: Fe
Anchoring strategy: Cystein-maleimide
Optimization: Chemical & genetic
Reaction: Lignin oxidation
Max TON: 20
ee: ---
PDB: ---
Notes: Reaction performed with a lignin model compound and hydrogen peroxide as oxidizing agent

Artificial Metalloenzymes for Hydrogenation and Transfer Hydrogenation Reactions

Review

Klein Gebbink, R. J. M.

Artificial Metalloenzymes and MetalloDNAzymes in Catalysis: From Design to Applications, 2018, 10.1002/9783527804085.ch6


Notes: Book chapter

Artificial Metalloenzymes on the Verge of New-to-Nature Metabolism

Review

Jeschek, M.

Trends Biotechnol., 2018, 10.1016/j.tibtech.2017.10.003


Notes: ---

Artificial Metalloenzymes: Reaction Scope and Optimization Strategies

Review

Lewis, J. C.; Ward, T. R.

Chem. Rev., 2018, 10.1021/acs.chemrev.7b00014


Notes: ---

Artificial Metalloproteins Containing Co4O4 Cubane Active Sites

Borovik, A. S.; Don Tilley, T.

J. Am. Chem. Soc., 2018, 10.1021/jacs.7b13052


Metal: Co
Ligand type: OAc; Pyridine
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Max TON: ---
ee: ---
PDB: 6AUC
Notes: Co-complex in Sav WT

Metal: Co
Ligand type: OAc; Pyridine
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Max TON: ---
ee: ---
PDB: 6AUE
Notes: Co-complex in Sav S112Y

A Whole Cell E. coli Display Platform for Artificial Metalloenzymes: Poly(phenylacetylene) Production with a Rhodium–Nitrobindin Metalloprotein

Schwaneberg, U.

ACS Catal., 2018, 10.1021/acscatal.7b04369


Metal: Rh
Ligand type: COD; Cp
Host protein: Nitrobindin variant NB4
Anchoring strategy: Cystein-maleimide
Optimization: ---
Max TON: 3046
ee: ---
PDB: ---
Notes: Calculated in vivo TON assuming 12800 metalloenzymes per E. coli cell

Capture and Characterization of a Reactive Haem– Carbenoid Complex in an Artificial Metalloenzyme

Hilvert, D.

Nat. Catal., 2018, 10.1038/s41929-018-0105-6


Metal: Fe
Host protein: Myoglobin (Mb)
Anchoring strategy: ---
Optimization: Genetic
Reaction: Cyclopropanation
Max TON: 1000
ee: 99
PDB: 6F17
Notes: Structure of the Mb*(NMH) haem-iron complex

Metal: Fe
Host protein: Myoglobin (Mb)
Anchoring strategy: ---
Optimization: Genetic
Reaction: Cyclopropanation
Max TON: 1000
ee: 99
PDB: 6G5B
Notes: Structure of the Mb*(NMH) haem-iron–carbenoid complex

Carbene in Cupredoxin Protein Scaffolds: Replacement of a Histidine Ligand in the Active Site Substantially Alters Copper Redox Properties

Albrecht, M.; Paradisi, F.

Angew. Chem., Int. Ed., 2018, 10.1002/ange.201807168


Metal: Cu
Host protein: Azurin
Anchoring strategy: Dative
Optimization: Chemical & genetic
Reaction: Electron transfer
Max TON: ---
ee: ---
PDB: ---
Notes: ---

Chimeric Streptavidins as Host Proteins for Artificial Metalloenzymes

Ward, T. R.; Woolfson, D. N.

ACS Catal., 2018, 10.1021/acscatal.7b03773


Metal: Ir
Ligand type: Cp*; Diamine
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Genetic
Max TON: 970
ee: 13
PDB: ---
Notes: ---

Metal: Ir
Ligand type: Cp*; Diamine
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Genetic
Max TON: 158
ee: 82
PDB: ---
Notes: ---

Metal: Ru
Ligand type: Carbene
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Genetic
Reaction: Olefin metathesis
Max TON: 105
ee: ---
PDB: ---
Notes: RCM, biotinylated Hoveyda-Grubbs second generation catalyst

Metal: ---
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Genetic
Reaction: Anion-π catalysis
Max TON: 6
ee: 41
PDB: ---
Notes: No metal

Design of Artificial Enzymes by Supramolecular Strategies

Review

Liu, J.

Curr. Opin. Struct. Biol., 2018, 10.1016/j.sbi.2018.02.003


Notes: ---

Development of De Novo Copper Nitrite Reductases: Where we are and where we need to go

Review

Pecoraro, V. L.

ACS Catal., 2018, 10.1021/acscatal.8b02153


Notes: ---

Directed Evolution of an Artificial Imine Reductase

Maréchal, J.-D.; Ward, T. R.

Angew. Chem., Int. Ed., 2018, 10.1002/anie.201711016


Metal: Ir
Ligand type: Amino-sulfonamide; Cp*
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Max TON: 380
ee: 95
PDB: 6ESS
Notes: Salsolidine formation; Sav mutant S112A-N118P-K121A-S122M: (R)-selective

Metal: Ir
Ligand type: Amino-sulfonamide; Cp*
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Max TON: 220
ee: 85
PDB: 6ESU
Notes: Salsolidine formation; Sav mutant S112R-N118P-K121A-S122M-L124Y: (S)-selective

Directed Evolution of Artificial Metalloenzymes: Bridging Synthetic Chemistry and Biology

Review

Arnold, F. H.

Artificial Metalloenzymes and MetalloDNAzymes in Catalysis: From Design to Applications, 2018, 10.1002/9783527804085.ch5


Notes: Book chapter

E. coli Surface Display of Streptavidin for Directed Evolution of an Allylic Deallylase

Ward, T. R.

Chem. Sci., 2018, 10.1039/c8sc00484f


Metal: Ru
Ligand type: Cp; Quinoline
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Genetic
Reaction: Deallylation
Max TON: 148
ee: ---
PDB: 6FH8
Notes: ---

Engineered Metalloenzymes with Non-Canonical Coordination Environments

Review

Green, A. P.; Hilvert, D.

Chem. - Eur. J., 2018, 10.1002/chem.201800975


Notes: ---

Evolving Artificial Metalloenzymes via Random Mutagenesis

Lewis, J. C.

Nat. Chem., 2018, 10.1038/nchem.2927


Metal: Rh
Ligand type: OAc
Anchoring strategy: Covalent
Optimization: Chemical & genetic
Reaction: Cyclopropanation
Max TON: 66
ee: 94
PDB: 5T88
Notes: Mutagenesis of the ArM by error-prone PCR

Metal: Rh
Ligand type: OAc
Anchoring strategy: Covalent
Optimization: Chemical & genetic
Reaction: N-H Insertion
Max TON: 73
ee: 40
PDB: 5T88
Notes: Mutagenesis of the ArM by error-prone PCR

Metal: Rh
Ligand type: OAc
Anchoring strategy: Covalent
Optimization: Chemical & genetic
Reaction: S-H Insertion
Max TON: 64
ee: 32
PDB: 5T88
Notes: Mutagenesis of the ArM by error-prone PCR

Metal: Rh
Ligand type: OAc
Anchoring strategy: Covalent
Optimization: Chemical & genetic
Reaction: Si-H Insertion
Max TON: 35
ee: 64
PDB: 5T88
Notes: Mutagenesis of the ArM by error-prone PCR

Ferritin Encapsulation of Artificial Metalloenzymes: Engineering a Tertiary Coordination Sphere for an Artificial Transfer Hydrogenase

Ward, T. R.

Dalton Trans., 2018, 10.1039/C8DT02224K


Metal: Ir
Ligand type: Amino-sulfonamide; Cp*
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Max TON: 3874
ee: 75
PDB: ---
Notes: ---

Functionalization of Protein Crystals with Metal Ions, Complexes and Nanoparticles

Review

Ueno, T.

Curr. Opin. Chem. Biol., 2018, 10.1016/j.cbpa.2017.11.015


Notes: ---

Generation of a Functional, Semisynthetic [FeFe]-Hydrogenase in a Photosynthetic Microorganism

Berggren, G.; Lindblad, P.

Energy Environ. Sci., 2018, 10.1039/C8EE01975D


Metal: Fe
Ligand type: CN; CO
Anchoring strategy: Reconstitution
Optimization: Chemical & genetic
Reaction: H2 evolution
Max TON: ---
ee: ---
PDB: ---
Notes: ---

Genetic Engineering of an Artificial Metalloenzyme for Transfer Hydrogenation of a Self-Immolative Substrate in Escherichia coli’s Periplasm

Ward, T. R.

J. Am. Chem. Soc., 2018, 10.1021/jacs.8b07189


Metal: Ir
Ligand type: Amino-sulfonamide; Cp*
Host protein: Streptavidin (Sav)
Anchoring strategy: Supramolecular
Optimization: Chemical & genetic
Max TON: 1000
ee: 76
PDB: 6GMI
Notes: ---

Going Beyond Structure: Nickel-Substituted Rubredoxin as a Mechanistic Model for the [NiFe] Hydrogenases

Shafaat, H. S.

J. Am. Chem. Soc., 2018, 10.1021/jacs.8b05194


Metal: Ni
Ligand type: Amino acid
Host protein: Rubredoxin (Rd)
Anchoring strategy: Metal substitution
Optimization: Genetic
Reaction: H2 evolution
Max TON: ---
ee: ---
PDB: ---
Notes: TOF = 149 s-1