Protein-Driven Electron-Transfer Process in a Fatty Acid Photodecarboxylase DOI Creative Commons
Giacomo Londi, Giacomo Salvadori, Patrizia Mazzeo

et al.

JACS Au, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 17, 2024

Naturally occurring photoenzymes are rare in nature, but among them, fatty acid photodecarboxylases derived from Chlorella variabilis (CvFAPs) have emerged as promising photobiocatalysts capable of performing the redox-neutral, light-induced decarboxylation free acids (FAs) into C1-shortened n-alka(e)nes. Using a hybrid QM/MM approach combined with polarizable embedding scheme, we identify structural changes active site and determine energetic landscape forward electron transfer (fET) FA substrate to excited flavin adenine dinucleotide. We obtain charge-transfer diradical structure where water molecule rearranges spontaneously form H-bond interaction flavin, while FA's carboxylate group twists migrates away it. Together, these modifications provide driving force necessary for fET proceed downhill direction. Moreover, by examining R451K mutant is farther core, show that marked reduction electronic coupling counterbalanced an increased force, resulting lifetime similar WT, thereby suggesting resilience process this mutation. Finally, through molecular dynamic simulations, reveal that, following fET, radical occurs within tens picoseconds, overcoming energy barrier ∼0.1 eV. Overall, providing atomistic characterization photoactivation CvFAP, work can be used future protein engineering.

Language: Английский

Directed Evolution and Unusual Protonation Mechanism of Pyridoxal Radical C–C Coupling Enzymes for the Enantiodivergent Photobiocatalytic Synthesis of Noncanonical Amino Acids DOI
Lei Cheng,

Zhiyu Bo,

Benjamin Krohn-Hansen

et al.

Journal of the American Chemical Society, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 23, 2025

Visible light-driven pyridoxal radical biocatalysis has emerged as a new strategy for the stereoselective synthesis of valuable noncanonical amino acids in protecting-group-free fashion. In our previously developed dehydroxylative C–C coupling using engineered PLP-dependent tryptophan synthases, an enzyme-controlled unusual α-stereochemistry reversal and pH-controlled enantiopreference were observed. Herein, through high-throughput photobiocatalysis, we evolved set stereochemically complementary PLP enzymes, allowing both l- d-amino with enhanced enantiocontrol across broad pH window. These newly acid synthases permitted use range organoboron substrates, including boronates, trifluoroborates, boronic acids, excellent efficiency. Mechanistic studies unveiled unexpected racemase activity earlier enzyme variants. This promiscuous was abolished shedding light on origin enantiocontrol. Further mechanistic investigations suggest switch proton donor to account stereoinvertive formation highlighting stereoinversion mechanism that is rare conventional two-electron enzymology.

Language: Английский

Citations

2
Asymmetric photoenzymatic incorporation of fluorinated motifs into olefins DOI
Mao‐Lin Li, Yaru Yuan, Wesley Harrison

et al.

Science, Journal Year: 2024, Volume and Issue: 385(6707), P. 416 - 421

Published: July 25, 2024

Enzymes capable of assimilating fluorinated feedstocks are scarce. This situation poses a challenge for the biosynthesis compounds used in pharmaceuticals, agrochemicals, and materials. We developed photoenzymatic hydrofluoroalkylation that integrates motifs into olefins. The photoinduced promiscuity flavin-dependent ene-reductases enables generation carbon-centered radicals from iodinated fluoroalkanes, which directed by photoenzyme to engage enantioselectively with approach facilitates stereocontrol through interaction between singular unit enzyme, securing high enantioselectivity at β, γ, or δ positions groups enzymatic hydrogen atom transfer-a process is notably challenging conventional chemocatalysis. work advances strategies integrating chemical opens avenues asymmetric synthesis compounds.

Language: Английский

Citations

17
Ground-state flavin-dependent enzymes catalyzed enantioselective radical trifluoromethylation DOI Creative Commons

Xinyu Duan,

Dong Cui,

Mengdi Wang

et al.

Nature Communications, Journal Year: 2025, Volume and Issue: 16(1)

Published: Jan. 31, 2025

Language: Английский

Citations

1
Threonine Aldolase-Catalyzed Enantioselective α-Alkylation of Amino Acids through Unconventional Photoinduced Radical Initiation DOI

Tian-Ci Wang,

Zheng Zhang, Guodong Rao

et al.

Journal of the American Chemical Society, Journal Year: 2024, Volume and Issue: 146(32), P. 22476 - 22484

Published: July 4, 2024

Visible light-driven pyridoxal radical biocatalysis has emerged as a promising strategy for the stereoselective synthesis of valuable noncanonical amino acids (ncAAs). Previously, use well-tailored photoredox catalysts represented key to enable efficient phosphate (PLP) enzyme-catalyzed reactions. Here, we report PLP-dependent threonine aldolase-catalyzed asymmetric α-C–H alkylation abundant using Katritzky pyridinium salts alkylating agents. The engineered aldolases allowed this redox-neutral proceed efficiently, giving rise challenging α-trisubstituted and -tetrasubstituted ncAA products in protecting-group-free fashion with excellent enantiocontrol. Mechanistically, enantioselective α-alkylation capitalizes on unique reactivity persistent enzymatic quinonoid intermediate derived from PLP cofactor acid substrate allow novel C–C coupling. Surprisingly, photobiocatalytic process does not require well-established operates through an unconventional photoinduced generation involving PLP-derived aldimine. ability develop reactions without relying classic photocatalysts or photoenzymes opens up new avenues advancing intermolecular that are known either organic chemistry enzymology.

Language: Английский

Citations

8
Photobiocatalysis: More than just an interesting lab curiosity? DOI Creative Commons
Yunjian Ma, Yutong Wang, Bin Wu

et al.

Chem Catalysis, Journal Year: 2024, Volume and Issue: 4(10), P. 101077 - 101077

Published: Aug. 23, 2024

Language: Английский

Citations

4
Merging photoredox with metalloenzymatic catalysis for enantioselective decarboxylative C(sp3)‒N3 and C(sp3)‒SCN bond formation DOI
Jinyan Rui,

Xinpeng Mu,

Jordi Soler Soler

et al.

Nature Catalysis, Journal Year: 2024, Volume and Issue: 7(12), P. 1394 - 1403

Published: Dec. 20, 2024

Language: Английский

Citations

4
Reshaping the Substrate-Binding Pocket of Ene-Reductase for Enhanced and Inverted Stereoselectivity: A Concise Access to the Stereocomplementary Chiral GABA Derivatives DOI

An Huang,

Xianheng Song,

Linlin Song

et al.

ACS Catalysis, Journal Year: 2025, Volume and Issue: 15(3), P. 1961 - 1972

Published: Jan. 20, 2025

Despite the availability of numerous natural and engineered ene-reductases (ERs), enantiocomplementary synthesis sterically hindered molecules by ERs is still limited poor substrate acceptance, particularly due to insufficient complementary stereoselectivity. Herein, we reshaped substrate-binding pocket SeER from Saccharomyces eubayanus through semirational design, enabling capable stereocomplementary hydrogenating challenging substituted β-cyano cinnamic esters. Compared wild type, variants exhibited enhanced activity (up 161-fold) catalytic efficiency kcat/KM 358-fold), displaying potential in synthesizing various chiral esters with high stereoselectivity 99% ee). Molecular dynamics simulations demonstrated that key for superior performance well-tuned pocket, which strengthens stabilizes recognition. Furthermore, elucidated practicality asymmetric GABA derivatives (e.g., Phenibut, Baclofen, Tolibut) via chemo-enzymatic cascade reactions.

Language: Английский

Citations

0
Enzymatic Synthesis of Saturated Bioisosteres of Ortho‐Substituted Benzenes by Artificial Photoenzyme DOI Open Access
Yuting Bai, Yi-yun Sheng, Yi Fu

et al.

Chemistry - A European Journal, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 17, 2025

Saturated bioisosteres of ortho-substituted benzenes are significant interest due to their enhanced pharmacokinetic properties, such as improved metabolic stability and reduced toxicity, making them valuable in drug design development. However, efficient synthesis remains a challenge organic chemistry. Herein, we report the biocatalytic saturated using engineered artificial photoenzymes. The photoenzyme, incorporating genetically encoded unnatural amino acids with benzophenone photosensitizer residue, facilitate formation chiral moderate enantiomeric excess via energy transfer process. Our results demonstrate versatility photoenzymes mediating new-to-nature reactions that difficult achieve conventional chemical or enzymatic methods.

Language: Английский

Citations

0
Mechanistic investigation of repurposed photoenzymes with new-to-nature reactivity DOI
Zhengyi Zhang, Mao‐Lin Li, Huimin Zhao

et al.

Current Opinion in Green and Sustainable Chemistry, Journal Year: 2025, Volume and Issue: 52, P. 101009 - 101009

Published: Feb. 27, 2025

Language: Английский

Citations

0
Catalytic Enantioselective Smiles Rearrangement Enabled by the Directed Evolution of P450 Radical Aryl Migratases DOI
Wen‐Zhen Fu,

Katherina Murcek,

Jasper Chen

et al.

Journal of the American Chemical Society, Journal Year: 2025, Volume and Issue: unknown

Published: April 1, 2025

Despite its synthetic potential, catalytic enantioselective Smiles rearrangement has remained elusive. Through the directed evolution of P450 radical aryl migratases (P450Smiles's), we describe first example rearrangement. A range racemic N-arylsulfonyl-α-chloroamides could be transformed by P450Smiles in an enantioconvergent manner, affording acyclic amide products possessing all-carbon quaternary stereocenter with excellent chemo- and enantioselectivity. Both electron-rich electron-deficient substituents were compatible migrating group, demonstrating this P450-catalyzed is insensitive to electronic properties group. Importantly, our evolved variants capable overriding innate cyclization activity N-alkyl amidyl intermediate, allowing chemoselective reductive formation products. Classical molecular dynamics (MD) simulations revealed unusual enzyme-controlled chemoselectivity stems from restricted conformation within enzyme active site, disfavoring pathway. This new-to-nature biocatalytic asymmetric showcases potential enzymatic enantioselectivity control over highly reactive intermediates eluding small-molecule catalysts.

Language: Английский

Citations

0