Engineering non-haem iron enzymes for enantioselective C(sp3)–F bond formation via radical fluorine transfer

Nature Synthesis, Journal Year: 2024, Volume and Issue: 3(8), P. 958 - 966

Published: March 28, 2024

Language: Английский

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Asymmetric photoenzymatic incorporation of fluorinated motifs into olefins

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

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Enzymatic Stereodivergent Access to Fluorinated β-Lactam Pharmacophores via Triple-Parameter Engineered Ketoreductases

ACS Catalysis, Journal Year: 2024, Volume and Issue: 14(8), P. 6358 - 6368

Published: April 11, 2024

Enzyme-catalyzed stereodivergent synthesis to access all possible stereoisomers of organofluorine compounds bearing multiple stereogenic centers remains an important and challenging subject. By integrative data-driven mining mechanism-guided engineering ketoreductases, we identified a biocatalytic platform produce four stereoisomeric fluoroalkyl amino acid esters two vicinal stereocenters. Fast triple-parameter coevolution via semirational CAST/ISM strategy provided the quadruple mutant M5 (A140K/L203T/G92A/V84I) ketoreductase BgADH not only displayed high stereoselectivity toward target (99:1 dr, 99% ee) but also observed with enhanced activity (kcat/Km, 6.3 folds) improved thermostability (T5015, 4 °C). Crystal structural analysis molecular dynamics (MD) simulation studies unveil residues (A140 F148) be key sites that are responsible for control stereoselectivity. The L203T/G92A mutation by affecting conformational distribution α-helix within active-site region, V84I thermal stability strengthening hydrogen bonding network neighboring residues. synthetic utility was further demonstrated substrate scope expansion, gram-scale reactions (648 g L–1 day–1), transformations chiral fluorinated β-lactams antibiotic carbapenem cores.

Language: Английский

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Recent updates on biodegradability and recyclability of bioplastics - Towards a new era in sustainability

Sustainable materials and technologies, Journal Year: 2024, Volume and Issue: 41, P. e01051 - e01051

Published: July 14, 2024

Language: Английский

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Role of the CrcB transporter of Pseudomonas putida in the multi‐level stress response elicited by mineral fluoride

Environmental Microbiology, Journal Year: 2022, Volume and Issue: 24(11), P. 5082 - 5104

Published: June 21, 2022

Abstract The presence of mineral fluoride (F − ) in the environment has both a geogenic and anthropogenic origin, halide been described to be toxic virtually all living organisms. While evidence gathered different microbial species supports this notion, systematic exploration effects F salts on metabolism physiology environmental bacteria remained underexplored thus far. In work, we studied characterized tolerance mechanisms deployed by model soil bacterium Pseudomonas putida KT2440 against NaF. By adopting systems‐level omic approaches, including functional genomics metabolomics, gauged impact anion at regulatory levels under conditions that impair bacterial growth. Several genes involved were isolated genome‐wide Tn ‐Seq screening—among which crcB , encoding an ‐specific exporter, was shown play predominant role detoxification. High‐resolution combined with assessment intracellular extracellular pH values quantitative experiments, underscored key nodes central carbon affected . Taken together, our results indicate P undergoes general, multi‐level stress response when challenged NaF significantly differs from caused other saline stressors. responses oxidative challenges have extensively literature, very little is known about metabolism. This state affairs contrasts fact more abundant than halides Earth crust (e.g. some soils, concentration can reach up 1 mg g −1 ). Understanding global treatment not only relevant unveil distinct detoxification but it could also guide engineering approaches for target incorporation fluorine into value‐added organofluorine molecules. regard, constitutes ideal explore such scenarios, since particularly its high level resistance variety physicochemical perturbations.

Language: Английский

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Mechanochemical synthesis of aromatic ketones: pyrylium tetrafluoroborate mediated deaminative arylation of amides

Chemical Science, Journal Year: 2024, Volume and Issue: 15(24), P. 9155 - 9163

Published: Jan. 1, 2024

A new method has been introduced that is able to tackle the complexities of N–C(O) activation in amide moieties through utilization pyrylium tetrafluoroborate a mechanochemical setting, where bonds undergo activation.

Language: Английский

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Nanotechnology-leveraged CRISPR/Cas systems: icebreaking in trace cancer-related nucleic acids biosensing

Molecular Cancer, Journal Year: 2025, Volume and Issue: 24(1)

Published: March 14, 2025

As promising noninvasive biomarkers, nucleic acids provide great potential to innovate cancer early detection methods and promote subsequent diagnosis improve the survival rates of patient. Accurate, straightforward sensitive such acid-based biomarkers in complex biological samples holds significant clinical importance. However, low abundance creates huge challenges for their routine detection. next-generation diagnostic tool, Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein (Cas) with high programmability, sensitivity, fidelity, single-base resolution, precise acid positioning capabilities are extremely attractive trace (NABCBs), permitting rapid, ultra-sensitive specific More importantly, by combing nanotechnology, it can solve long-lasting problems poor accuracy simplicity, as well achieve integrated miniaturization portable point-of-care testing (POCT) existing literature lacks emphasis on this topic. Thus, we intend propose a timely one readers. This review will bridge gap providing insights CRISPR/Cas-based nano-biosensing development highlighting current state-of-art, challenges, prospects. We expect that better understanding valuable NABCBs detection, thereby facilitating advancements screening/detection/diagnostics win practical applications foreseeable future.

Language: Английский

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Synthetic metabolism for biohalogenation

Current Opinion in Biotechnology, Journal Year: 2021, Volume and Issue: 74, P. 180 - 193

Published: Dec. 23, 2021

The pressing need for novel bioproduction approaches faces a limitation in the number and type of molecules accessed through synthetic biology. Halogenation is widely used tuning physicochemical properties polymers, but traditional halogenation chemistry often lacks specificity generates harmful by-products. Here, we pose that deploying metabolism tailored biohalogenation represents an unique opportunity towards economically attractive environmentally friendly organohalide production. On this background, discuss growth-coupled selection functional metabolic modules harness rich repertoire biosynthetic biodegradation capabilities environmental bacteria vivo biohalogenation. By rationally combining these approaches, chemical landscape living cells can accommodate added-value organohalides which, as today, are obtained by chemistry.

Language: Английский

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Chemoenzymatic Synthesis of Fluorinated Mycocyclosin Enabled by the Engineered Cytochrome P450-Catalyzed Biaryl Coupling Reaction

Journal of the American Chemical Society, Journal Year: 2024, Volume and Issue: 146(29), P. 19962 - 19973

Published: July 10, 2024

Installing fluorine atoms onto natural products holds great promise for the generation of fluorinated molecules with improved or novel pharmacological properties. The enzymatic oxidative carbon-carbon coupling reaction represents a straightforward strategy synthesizing biaryl architectures, but exploration this method producing fluorine-substituted derivatives remains elusive. Here, in study, we report protein engineering cytochrome P450 from Mycobacterium tuberculosis (MtCYP121) construction series new-to-nature Mycocyclosin derivatives. This protocol takes advantage "hybrid" chemoenzymatic procedure consisting tyrosine phenol lyase-catalyzed fluorotyrosine preparation commercially available fluorophenols, intermolecular chemical condensation to give cyclodityrosines, and an engineered MtCYP121-catalyzed intramolecular biphenol complete strained macrocyclic structure. Computational mechanistic studies reveal that MtCYP121 employs Cpd I abstract hydrogen atom proximal phenolic hydroxyl group substrate trigger reaction. Then, conformational change makes two groups close enough undergo transfer assistance pocket water molecule. final diradical process completes C-C bond formation. efficiency was found be influenced by various substitutions, primarily due presence distinct binding conformations.

Language: Английский

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Challenges and opportunities in bringing nonbiological atoms to life with synthetic metabolism

Trends in biotechnology, Journal Year: 2022, Volume and Issue: 41(1), P. 27 - 45

Published: June 30, 2022

The relatively narrow spectrum of chemical elements within the microbial 'biochemical palate' limits reach biotechnology, because several added-value compounds can only be produced with traditional organic chemistry. Synthetic biology offers enabling tools to tackle this issue by facilitating 'biologization' non-canonical atoms. interplay between xenobiology and synthetic metabolism multiplies routes for incorporating nonbiological atoms into engineered microbes. In review, we survey natural assimilation beyond essential [i.e., carbon (C), hydrogen (H), nitrogen (N), oxygen (O), phosphorus (P), sulfur (S)], discussing how these mechanisms could repurposed biotechnology. Furthermore, propose a computational framework identify amenable biologization, ranking reactions suitable build metabolism. When combined deployed in robust hosts, approaches will offer sustainable alternatives smart production.

Language: Английский

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