14 publications
-
Addressable DNA–Myoglobin Photocatalysis
-
Chem. - Asian J. 2009, 4, 1064-1069, 10.1002/asia.200900082
A hybrid myoglobin, containing a single‐stranded DNA anchor and a redox‐active ruthenium moiety tethered to the heme center can be used as a photocatalyst. The catalyst can be selectively immobilized on a surface‐bound complementary DNA molecule and thus readily recycled from complex reaction mixtures. This principle may be applied to a range of heme‐dependent enzymes allowing the generation of novel light‐triggered photocatalysts. Photoactivatable myoglobin containing a DNA oligonucleotide as a structural anchor was designed by using the reconstitution of artificial heme moieties containing Ru3+ ions. This semisynthetic DNA–enzyme conjugate was successfully used for the oxidation of peroxidase substrates by using visible light instead of H2O2 for the activation. The DNA anchor was utilized for the immobilization of the enzyme on the surface of magnetic microbeads. Enzyme activity measurements not only indicated undisturbed biofunctionality of the tethered DNA but also enabled magnetic separation‐based enrichment and recycling of the photoactivatable biocatalyst.
Metal: RuLigand type: BipyridineHost protein: Myoglobin (Mb)Anchoring strategy: SupramolecularOptimization: ---Notes: Horse heart myoglobin
-
Aqueous Oxidation of Alcohols Catalyzed by Artificial Metalloenzymes Based on the Biotin–Avidin Technology
-
J. Organomet. Chem. 2005, 690, 4488-4491, 10.1016/j.jorganchem.2005.02.001
Based on the incorporation of biotinylated organometallic catalyst precursors within (strept)avidin, we have developed artificial metalloenzymes for the oxidation of secondary alcohols using tert-butylhydroperoxide as oxidizing agent. In the presence of avidin as host protein, the biotinylated aminosulfonamide ruthenium piano stool complex 1 (0.4 mol%) catalyzes the oxidation of sec-phenethyl alcohol at room temperature within 90 h in over 90% yield. Gel electrophoretic analysis of the reaction mixture suggests that the host protein is not oxidatively degraded during catalysis.
Metal: RuHost protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
Metal: RuHost protein: Avidin (Av)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
Metal: RuHost protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
Metal: RhHost protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
Metal: IrHost protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
-
Atroposelective Antibodies as a Designed Protein Scaffold for Artificial Metalloenzymes
-
Sci. Rep. 2019, 9, 10.1038/s41598-019-49844-0
Design and engineering of protein scaffolds are crucial to create artificial metalloenzymes. Herein we report the first example of C-C bond formation catalyzed by artificial metalloenzymes, which consist of monoclonal antibodies (mAbs) and C2 symmetric metal catalysts. Prepared as a tailored protein scaffold for a binaphthyl derivative (BN), mAbs bind metal catalysts bearing a 1,1?-bi-isoquinoline (BIQ) ligand to yield artificial metalloenzymes. These artificial metalloenzymes catalyze the Friedel-Crafts alkylation reaction. In the presence of mAb R44E1, the reaction proceeds with 88% ee. The reaction catalyzed by Cu-catalyst incorporated into the binding site of mAb R44E1 is found to show excellent enantioselectivity with 99% ee. The protein environment also enables the use of BIQ-based catalysts as asymmetric catalysts for the first time.
Notes: ---
-
Autoxidation of Ascorbic Acid Catalyzed by a Semisynthetic Enzyme
-
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).
Metal: CuLigand type: BipyridineHost protein: Papain (PAP)Anchoring strategy: CovalentOptimization: ---Notes: ---
-
Bimetallic Copper-Heme-Protein-DNA Hybrid Catalyst for Diels Alder Reaction
-
Croat. Chem. Acta 2011, 84, 269-275, 10.5562/cca1828
A bimetallic heme-DNA cofactor, containing an iron and a copper center, was synthesized for the design of novel hybrid catalysts for stereoselective synthesis. The cofactor was used for the reconstitution of apo-myoglobin. Both the cofactor alone and its myoglobin adduct were used to catalyze a model Diels Alder reaction. Stereoselectivity of this conversion was analyzed by chiral HPLC. Reactions carried out in the presence of myoglobin-heme-Cu-DNA catalyst showed greater product conversion and stereoselectivity than those carried out with the heme-Cu-DNA cofactor. This observation suggested that the protein shell plays a significant role in the catalytic conversion.
Metal: CuLigand type: BipyridineHost protein: Myoglobin (Mb)Anchoring strategy: SupramolecularOptimization: ---Notes: Horse heart myoglobin
-
Biosynthesis of a Site-Specific DNA Cleaving Protein
-
J. Am. Chem. Soc. 2008, 130, 13194-13195, 10.1021/ja804653f
An E. coli catabolite activator protein (CAP) has been converted into a sequence-specific DNA cleaving protein by genetically introducing (2,2′-bipyridin-5-yl)alanine (Bpy-Ala) into the protein. The mutant CAP (CAP-K26Bpy-Ala) showed comparable binding affinity to CAP-WT for the consensus operator sequence. In the presence of Cu(II) and 3-mercaptopropionic acid, CAP-K26Bpy-Ala cleaves double-stranded DNA with high sequence specificity. This method should provide a useful tool for mapping the molecular details of protein−nucleic acid interactions.
Metal: CuLigand type: BipyridineHost protein: Catabolite activator protein (CAP)Anchoring strategy: ---Optimization: Chemical & geneticNotes: Catabolite activator protein from E. coli
Metal: FeLigand type: BipyridineHost protein: Catabolite activator protein (CAP)Anchoring strategy: ---Optimization: Chemical & geneticNotes: Catabolite activator protein from E. coli
-
Design of an Enantioselective Artificial Metallo-Hydratase Enzyme Containing an Unnatural Metal-Binding Amino Acid
-
Chem. Sci. 2017, 8, 7228-7235, 10.1039/C7SC03477F
The design of artificial metalloenzymes is a challenging, yet ultimately highly rewarding objective because of the potential for accessing new-to-nature reactions. One of the main challenges is identifying catalytically active substrate–metal cofactor–host geometries. The advent of expanded genetic code methods for the in vivo incorporation of non-canonical metal-binding amino acids into proteins allow to address an important aspect of this challenge: the creation of a stable, well-defined metal-binding site. Here, we report a designed artificial metallohydratase, based on the transcriptional repressor lactococcal multidrug resistance regulator (LmrR), in which the non-canonical amino acid (2,2′-bipyridin-5yl)alanine is used to bind the catalytic Cu(II) ion. Starting from a set of empirical pre-conditions, a combination of cluster model calculations (QM), protein–ligand docking and molecular dynamics simulations was used to propose metallohydratase variants, that were experimentally verified. The agreement observed between the computationally predicted and experimentally observed catalysis results demonstrates the power of the artificial metalloenzyme design approach presented here.
Notes: ---
-
Dual Modification of a Triple-Stranded β-Helix Nanotube with Ru and Re Metal Complexes to Promote Photocatalytic Reduction of CO2
-
Chem. Commun. 2011, 47, 2074, 10.1039/C0CC03015E
We have constructed a robust β-helical nanotube from the component proteins of bacteriophage T4 and modified this nanotube with RuII(bpy)3 and ReI(bpy)(CO)3Cl complexes. The photocatalytic system arranged on the tube catalyzes the reduction of CO2 with higher reactivity than that of the mixture of the monomeric forms.
Notes: ---
Metal: RuLigand type: BipyridineHost protein: [(gp5βf)3]2Anchoring strategy: Lysine-succinimideOptimization: GeneticNotes: ---
-
Enantioselective Artificial Metalloenzymes by Creation of a Novel Active Site at the Protein Dimer Interface
-
Angew. Chem. Int. Ed. 2012, 51, 7472-7475, 10.1002/anie.201202070
A game of two halves: Artificial metalloenzymes are generated by forming a novel active site on the dimer interface of the transcription factor LmrR. Two copper centers are incorporated by binding to ligands in each half of the dimer. With this system up to 97 % ee was obtained in the benchmark CuII catalyzed Diels–Alder reaction (see scheme).
-
Expanding the Chemical Diversity in Artificial Imine Reductases Based on the Biotin–Streptavidin Technology
-
ChemCatChem 2014, 6, 1010-1014, 10.1002/cctc.201300825
We report on the optimization of an artificial imine reductase based on the biotin‐streptavidin technology. With the aim of rapidly generating chemical diversity, a novel strategy for the formation and evaluation of biotinylated complexes is disclosed. Tethering the biotin‐anchor to the Cp* moiety leaves three free coordination sites on a d6 metal for the introduction of chemical diversity by coordination of a variety of ligands. To test the concept, 34 bidentate ligands were screened and a selection of the 6 best was tested in the presence of 21 streptavidin (Sav) isoforms for the asymmetric imine reduction by the resulting three legged piano stool complexes. Enantiopure α‐amino amides were identified as promising bidentate ligands: up to 63 % ee and 190 turnovers were obtained in the formation of 1‐phenyl‐1,2,3,4‐tetrahydroisoquinoline with [IrCp*biotin(L‐ThrNH2)Cl]⊂SavWT as a catalyst.
Metal: IrHost protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
Metal: IrHost protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
Metal: IrHost protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
Metal: IrLigand type: Cp*Host protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
Metal: IrHost protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
Metal: IrHost protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
Metal: IrHost protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
Metal: IrHost protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
Metal: IrHost protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
Metal: RhHost protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
Metal: RhHost protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
Metal: RhHost protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
Metal: RhLigand type: Cp*Host protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
Metal: RhHost protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
Metal: RhHost protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
Metal: RhHost protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
Metal: RhHost protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
-
Novel Artificial Metalloenzymes by In Vivo Incorporation of Metal-Binding Unnatural Amino Acids
-
Chem. Sci. 2015, 6, 770-776, 10.1039/c4sc01525h
Artificial metalloenzymes have emerged as an attractive new approach to enantioselective catalysis. Herein, we introduce a novel strategy for preparation of artificial metalloenzymes utilizing amber stop codon suppression methodology for the in vivo incorporation of metal-binding unnatural amino acids. The resulting artificial metalloenzymes were applied in catalytic asymmetric Friedel–Crafts alkylation reactions and up to 83% ee for the product was achieved.
Notes: ---
-
Photo-Driven Hydrogen Evolution by an Artificial Hydrogenase Utilizing the Biotin-Streptavidin Technology
-
Helv. Chim. Acta 2018, 101, e1800036, 10.1002/hlca.201800036
Photocatalytic hydrogen evolution by an artificial hydrogenase based on the biotin‐streptavidin technology is reported. A biotinylated cobalt pentapyridyl‐based hydrogen evolution catalyst (HEC) was incorporated into different mutants of streptavidin. Catalysis with [Ru(bpy)3]Cl2 as a photosensitizer (PS) and ascorbate as sacrificial electron donor (SED) at different pH values highlighted the impact of close lying amino acids that may act as a proton relay under the reaction conditions (Asp, Arg, Lys). In the presence of a close‐lying lysine residue, both, the rates were improved, and the reaction was initiated much faster. The X‐ray crystal structure of the artificial hydrogenase reveals a distance of 8.8 Å between the closest lying Co‐moieties. We thus suggest that the hydrogen evolution mechanism proceeds via a single Co centre. Our findings highlight that streptavidin is a versatile host protein for the assembly of artificial hydrogenases and their activity can be fine‐tuned via mutagenesis.
Metal: CoHost protein: Streptavidin (Sav)Anchoring strategy: SupramolecularOptimization: Chemical & geneticNotes: ---
-
Semi-Synthesis of an Artificial Scandium(III) Enzyme with a β-Helical Bio-Nanotube
-
Dalton Trans. 2012, 41, 11424, 10.1039/C2DT31030A
We have succeeded in preparing semi-synthesized proteins bound to Sc3+ ion which can promote an epoxide ring-opening reaction. The Sc3+ binding site was created on the surface of [(gp5βf)3]2 (N. Yokoi et al., Small, 2010, 6, 1873) by introducing a cysteine residue for conjugation of a bpy moiety using a thiol–maleimide coupling reaction. Three cysteine mutants [(gp5βf_X)3]2 (X = G18C, L47C, N51C) were prepared to introduce a bpy in different positions because it had been reported that Sc3+ ion can serve as a Lewis-acid catalyst for an epoxide ring-opening reaction upon binding of epoxide to bpy and two –ROH groups. G18C_bpy with Sc3+ can accelerate the rate of catalysis of the epoxide ring-opening reaction and has the highest rate of conversion among the three mutants. The value is more than 20 times higher than that of the mixtures of [(gp5βf)3]2/2,2′-bipyridine and L-threonine/2,2′-bipyridine. The elevated activity was obtained by the cooperative effect of stabilizing the Sc3+ coordination and accumulation of substrates on the protein surface. Thus, we expect that the semi-synthetic approach can provide insights into new rational design of artificial metalloenzymes.
Metal: ScLigand type: BipyridineHost protein: [(gp5βf)3]2Anchoring strategy: Cystein-maleimideOptimization: GeneticNotes: ---
-
Semisynthesis of Bipyridyl-Alanine Cytochrome c Mutants: Novel Proteins with Enhanced Electron-Transfer Properties
-
J. Am. Chem. Soc. 1993, 115, 8455-8456, 10.1021/ja00071a068
n/a
Notes: No catalysis