Engineered and artificial metalloenzymes for selective C–H functionalization

Ren, Xinkun; Fasan, Rudi

doi:10.1016/j.cogsc.2021.100494

Cited by 59 publications

(41 citation statements)

References 118 publications

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“…Previous efforts by our group and others have led to the development of hemoprotein-based biocatalysts for a growing number of abiological carbene transfer reactions, − including carbene insertions into heteroatom–hydrogen bonds (N–H, S–H, Si–H, and B–H). − In particular, engineered myoglobins, P450 enzymes, and artificial metalloenzymes have been reported for (nonasymmetric) N–H insertion reactions involving acceptor-only diazo esters. ,,,, More recently, the first example of a biocatalytic asymmetric N–H insertion with donor–acceptor α-alkyl-substituted diazo esters was accomplished . On the other hand, laboratory-evolved variants of cytochrome c from Rhodothermus marinus have proven useful for stereoselective Si–H and B–H , insertion reactions in the presence of donor–acceptor diazo compounds.…”

Section: Introductionmentioning

confidence: 99%

Enantioselective Synthesis of α-Trifluoromethyl Amines via Biocatalytic N–H Bond Insertion with Acceptor-Acceptor Carbene Donors

et al. 2022

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The biocatalytic toolbox has recently been expanded to include enzyme-catalyzed carbene transfer reactions not occurring in Nature. Herein, we report the development of a biocatalytic strategy for the synthesis of enantioenriched α-trifluoromethyl amines through an asymmetric N–H carbene insertion reaction catalyzed by engineered variants of cytochrome c 552 from Hydrogenobacter thermophilus . Using a combination of protein and substrate engineering, this metalloprotein scaffold was redesigned to enable the synthesis of chiral α-trifluoromethyl amino esters with up to >99% yield and 95:5 er using benzyl 2-diazotrifluoropropanoate as the carbene donor. When the diazo reagent was varied, the enantioselectivity of the enzyme could be inverted to produce the opposite enantiomers of these products with up to 99.5:0.5 er. This methodology is applicable to a broad range of aryl amine substrates, and it can be leveraged to obtain chemoenzymatic access to enantioenriched β-trifluoromethyl-β-amino alcohols and halides. Computational analyses provide insights into the interplay of protein- and reagent-mediated control on the enantioselectivity of this reaction. This work introduces the first example of a biocatalytic N–H carbenoid insertion with an acceptor–acceptor carbene donor, and it offers a biocatalytic solution for the enantioselective synthesis of α-trifluoromethylated amines as valuable synthons for medicinal chemistry and the synthesis of bioactive molecules.

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Section: Introductionmentioning

confidence: 99%

Enantioselective Synthesis of α-Trifluoromethyl Amines via Biocatalytic N–H Bond Insertion with Acceptor-Acceptor Carbene Donors

et al. 2022

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“…Enzymatic C–H bond functionalization is an effective strategy for building and diversifying organic molecules. − C–H hydroxylation, an important C–H functionalization reaction, is an attractive pathway for synthesizing hydroxy amino acids, which are widely used as chiral building blocks in the pharmaceutical and fine chemical industries. For example, trans -4-hydroxy- l -proline is the functional intermediate for the synthesis of N -alkylpyrrole and carbapenem antibiotics. , Moreover, a series of hydrophobic hydroxy amino acid natural products from legume seeds exhibit important physiological activities; for example, (2 S ,3 R ,4 S )-hydroxyisoleucine (4-HIL) is a natural nonproteinogenic amino acid derived from fenugreek seeds with potential insulinotropic biological activity …”

Section: Introductionmentioning

confidence: 99%

Molecular Insights into the Regioselectivity of the Fe(II)/2-Ketoglutarate-Dependent Dioxygenase-Catalyzed C–H Hydroxylation of Amino Acids

Jing

et al. 2022

ACS Catal.

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l-Isoleucine dioxygenase (IDO) directly catalyzes the C–H bond hydroxylation of several hydrophobic aliphatic amino acids. However, the ambiguous selectivity of IDO prevents its application in chiral hydroxy amino acid production. The hydroxylation of l-norleucine by IDO, which produces 4-hydroxynorleucine and 5-hydroxynorleucine with obvious regioselectivity, was used to investigate the mechanism of IDO regioselectivity. Along with computational structural analysis and high-throughput screening, the IDO structure revealed single-site variants (T244A, T244G, and T244S) with enhanced regioselectivity; for example, the 4-hydroxynorleucine purity in regioisomeric products increased from 78.9% (by wild-type IDO) to 95.1%, 96.6%, and 95.3%, respectively. Molecular dynamics simulations showed that mutating T244 into smaller amino acids fine-tuned the substrate binding pose. For asymmetric catalysis requiring precise positioning, this change expanded the most frequent distances between the substrate C4 or C5 and Fe2+, giving a maximum 4-hydroxynorleucine purity of 96.6%. We improved the understanding of the regioselectivity of Fe(II)/2-ketoglutarate-dependent dioxygenases and provide a route for diversifying C–H hydroxylation-based active compounds.

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“… 22 , 23 Repurposed (natural) enzymes and ArMs offer an attractive means to complement SAM-dependent enzymes for C–H activation purposes. 21 , 24 − 28 Among the most noteworthy achievements in “new-to-nature” C–C bond formation via C–H functionalization, one should mention engineered P411, 29 , 30 repurposed P450, 31 − 34 myoglobin, 35 , 36 and streptavidin 37 , 38 ArMs. However, examples of highly active C–H insertion biocatalysts that are based on first-row transition metals and tolerate aerobic reaction conditions remain scarce.…”

Section: Introductionmentioning

confidence: 99%

An Artificial Metalloenzyme Based on a Copper Heteroscorpionate Enables sp³ C–H Functionalization via Intramolecular Carbene Insertion

et al. 2022

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The selective functionalization of sp 3 C–H bonds is a versatile tool for the diversification of organic compounds. Combining attractive features of homogeneous and enzymatic catalysts, artificial metalloenzymes offer an ideal means to selectively modify these inert motifs. Herein, we report on a copper(I) heteroscorpionate complex embedded within streptavidin that catalyzes the intramolecular insertion of a carbene into sp 3 C–H bonds. Target residues for genetic optimization of the artificial metalloenzyme were identified by quantum mechanics/molecular mechanics simulations. Double-saturation mutagenesis yielded detailed insight on the contribution of individual amino acids on the activity and the selectivity of the artificial metalloenzyme. Mutagenesis at a third position afforded a set of artificial metalloenzymes that catalyze the enantio- and regioselective formation of β- and γ-lactams with high turnovers and promising enantioselectivities.

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Engineered and artificial metalloenzymes for selective C–H functionalization

Cited by 59 publications

References 118 publications

Enantioselective Synthesis of α-Trifluoromethyl Amines via Biocatalytic N–H Bond Insertion with Acceptor-Acceptor Carbene Donors

Enantioselective Synthesis of α-Trifluoromethyl Amines via Biocatalytic N–H Bond Insertion with Acceptor-Acceptor Carbene Donors

Molecular Insights into the Regioselectivity of the Fe(II)/2-Ketoglutarate-Dependent Dioxygenase-Catalyzed C–H Hydroxylation of Amino Acids

An Artificial Metalloenzyme Based on a Copper Heteroscorpionate Enables sp³ C–H Functionalization via Intramolecular Carbene Insertion

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Engineered and artificial metalloenzymes for selective C–H functionalization

Cited by 59 publications

References 118 publications

Enantioselective Synthesis of α-Trifluoromethyl Amines via Biocatalytic N–H Bond Insertion with Acceptor-Acceptor Carbene Donors

Enantioselective Synthesis of α-Trifluoromethyl Amines via Biocatalytic N–H Bond Insertion with Acceptor-Acceptor Carbene Donors

Molecular Insights into the Regioselectivity of the Fe(II)/2-Ketoglutarate-Dependent Dioxygenase-Catalyzed C–H Hydroxylation of Amino Acids

An Artificial Metalloenzyme Based on a Copper Heteroscorpionate Enables sp3 C–H Functionalization via Intramolecular Carbene Insertion

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An Artificial Metalloenzyme Based on a Copper Heteroscorpionate Enables sp³ C–H Functionalization via Intramolecular Carbene Insertion