A selective and atom-economic rearrangement of uridine by cascade biocatalysis for production of pseudouridine

Nature Communications, Journal Year: 2023, Volume and Issue: 14(1)

Published: April 20, 2023

As a crucial factor of their therapeutic efficacy, the currently marketed mRNA vaccines feature uniform substitution uridine (U) by corresponding C-nucleoside, pseudouridine (Ψ), in 1-N-methylated form. Synthetic supply building block (1-N-Me-Ψ-5'-triphosphate) involves expedient access to Ψ as principal challenge. Here, we show selective and atom-economic 1N-5C rearrangement β-D-ribosyl on uracil obtain from unprotected U quantitative yield. One-pot cascade transformation four enzyme-catalyzed steps, via D-ribose (Rib)-1-phosphate, Rib-5-phosphate (Rib5P) Ψ-5'-phosphate (ΨMP), gives Ψ. Coordinated function coupled enzymes overall necessitates specific release phosphate ΨMP, but not intermediary ribose phosphates. Discovery Yjjg ΨMP-specific phosphatase enables internally controlled regeneration catalytic reagent. With driving force provided net N-C rearrangement, optimized reaction yields supersaturated product solution (∼250 g/L) which pure crystallizes (90% recovery). Scale up 25 g isolated at enzyme turnovers ∼105 mol/mol demonstrates robust process technology, promising for production. Our study identifies multistep reaction, realized biocatalysis, C-nucleoside synthesis high efficiency.

Language: Английский

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Stereodivergent Protein Engineering of Fatty Acid Photodecarboxylase for Light‐Driven Kinetic Resolution of Sec‐Alcohol Oxalates

Angewandte Chemie International Edition, Journal Year: 2024, Volume and Issue: 63(10)

Published: Jan. 10, 2024

Stereodivergent engineering of one enzyme to create stereocomplementary variants for synthesizing optically pure molecules with tailor-made (R) or (S) configurations on an optional basis is highly desirable and challenging. This study aimed engineer fatty acid photodecarboxylase from Chlorella variabilis (CvFAP) using the focused rational iterative site-specific mutagenesis (FRISM) strategy obtain two excellent selectivity (both giving products up 99 % e.e.). These were used CvFAP-catalyzed light-driven kinetic resolution oxalates oxamic acids prepared corresponding sec-alcohols amines, providing a new biotransformation process preparing chiral amines. Molecular dynamics simulation, data transient spectra revealed source selectivity. represents first example amines catalyzed by pair CvFAPs.

Language: Английский

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Artificial Photosynthesis for Regioselective Reduction of NAD(P)⁺ to NAD(P)H Using Water as an Electron and Proton Source

Journal of the American Chemical Society, Journal Year: 2024, Volume and Issue: 146(8), P. 5152 - 5161

Published: Feb. 13, 2024

In photosynthesis, four electrons and protons taken from water in photosystem II (PSII) are used to reduce NAD(P)+ produce NAD(P)H I (PSI), which is the most important reductant CO2. Despite extensive efforts mimic artificial photosynthesis using electron proton sources has yet be achieved. Herein, we report photocatalytic reduction of its analogues a molecular model PSI, combined with oxidation PSII. Photoirradiation toluene/trifluoroethanol (TFE)/borate buffer aqueous solution hydroquinone derivatives (X-QH2), 9-mesityl-10-methylacridinium ion, cobaloxime, (PSI model) resulted quantitative regioselective formation p-benzoquinone (X-Q). X-Q was reduced X-QH2, accompanied by dioxygen under photoirradiation toluene/TFE/borate [(N4Py)FeII]2+ (PSII model). The PSI PSII models were two glass membranes liquid as an source turnover number (TON) 54. To best our knowledge, this first time achieve stoichiometry O2.

Language: Английский

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Biocatalytic Heteroaromatic Amide Formation in Water Enabled by a Catalytic Tetrad and Two Access Tunnels

ACS Catalysis, Journal Year: 2024, Volume and Issue: 14(11), P. 8913 - 8921

Published: May 25, 2024

The amide moiety belongs to the most common motives in pharmaceutical chemistry, present many prescribed small-molecule pharmaceuticals. Methods for its manufacture are still high demand, especially using water/buffer as a solvent and avoiding stoichiometric amounts of activation reagents. Herein, we identified from library lipases/esterases/acyltransferases variants thereof lipase originating Sphingomonas sp. HXN-200 (SpL) able form amides aqueous solution starting broad scope sterically demanding heteroaromatic ethyl esters well aliphatic amines, reaching isolated yields up 99% on preparative scale space time 864 g L–1 d–1; thus, selected cases, was formed within minutes. enzyme features an aspartate next canonical serine catalytic triad, which essential formation. Furthermore, structure revealed two tunnels active site, presumably one ester amine, permit bringing together amine site. This work shows that biocatalytic formation various five- six-membered buffer can serve platform synthesis.

Language: Английский

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Peptide and Enzyme Catalysts Work in Concert in Stereoselective Cascade Reactions—Oxidation followed by Conjugate Addition

Angewandte Chemie International Edition, Journal Year: 2024, Volume and Issue: 63(12)

Published: Jan. 18, 2024

Abstract Enzymes and peptide catalysts consist of the same building blocks but require vastly different environments to operate best. Herein, we show that an enzyme a catalyst can work together in single reaction vessel catalyze two‐step cascade with high chemo‐ stereoselectivity. Abundant linear alcohols, nitroolefins, alcohol oxidase, tripeptide provided chiral γ‐nitroaldehydes aqueous buffer. High yields (up 92 %) stereoselectivities 98 % ee) were achieved for through rational design identification common conditions.

Language: Английский

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