Β-keto Acids Research Articles

Synergistic Catalytic Asymmetric Decarboxylation of β-Keto Acids with Acyclic α-Arylenamides

Synergistic Catalytic Asymmetric Decarboxylation of β-Keto Acids with Acyclic α-Arylenamides

Phytochemical Volatiles as Potential Bionematicides with Safer Ecotoxicological Properties.

The management of plant-parasitic nematodes (PPNs) still relies on traditional nematicides that threaten the environment and human health. Novel solutions are urgently needed for PPN pest management that are effective while safeguarding non-target organisms. Volatile phytochemicals belong to a structurally diverse group of bioactive metabolites that are believed to hold safer environmental characteristics than synthetic pesticides. Nonetheless, not many studies have analysed the potential environmental benefits of shifting to these novel bionematicides. In the present study, 20 phytochemical volatiles with reported nematicidal activity were compared to traditional pesticides using specific parameters of environmental and human health safety available on applied online databases and predicted in silico through specialised software. Overall, the reviewed nematicidal phytochemicals were reportedly less toxic than synthetic nematicides. They were predicted to disperse to the air and soil environmental compartments and were reported to have a lower toxicity on aquatic organisms. On the contrary, the synthetic nematicides were reportedly toxic to aquatic organisms while showing a predicted high affinity to the water environmental compartment. As alternatives, β-keto or fatty acid derivatives, e.g., aliphatic alcohols or ketones, showed more adequate properties. This study highlights the importance of complementing studies on nematicidal activity with a risk assessment-based analysis to allow for a faster selection of nematicidal phytochemical volatiles and to leverage the development and implementation of bionematicides.

Molecular and Structural Basis for Cγ-C Bond Formation by PLP-Dependent Enzyme Fub7.

Pyridoxal 5'-phosphate (PLP)-dependent enzymes that catalyze γ-replacement reactions are prevalent, yet their utilization of carbon nucleophile substrates is rare. The recent discovery of two PLP-dependent enzymes, CndF and Fub7, has unveiled unique C-C bond forming capabilities, enabling the biocatalytic synthesis of alkyl- substituted pipecolic acids from O-acetyl-L-homoserine and β-keto acid or aldehyde derived enolates. This breakthrough presents fresh avenues for the biosynthesis of pipecolic acid derivatives. However, the catalytic mechanisms of these enzymes remain elusive, and a dearth of structural information hampers their extensive application. Here, we have broadened the catalytic scope of Fub7 by employing ketone-derived enolates as carbon nucleophiles, revealing Fub7's capacity for substrate-dependent regioselective α-alkylation of unsymmetrical ketones. Through an integrated approach combining X-ray crystallography, spectroscopy, mutagenesis, and computational docking studies, we offer a detailed mechanistic insight into Fub7 catalysis. Our findings elucidate the structural basis for its substrate specificity, stereoselectivity, and regioselectivity. Our work sets the stage ready for subsequent protein engineering effort aimed at expanding the synthetic utility of Fub7, potentially unlocking novel methods to access a broader array of noncanonical amino acids.

An acyl-CoA thioesterase is essential for the biosynthesis of a key dauer pheromone in C. elegans

Methyl ketone (MK)-ascarosides represent essential components of several pheromones in Caenorhabditis elegans, including the dauer pheromone, which triggers the stress-resistant dauer larval stage, and the male-attracting sex pheromone. Here, we identify an acyl-CoA thioesterase, ACOT-15, that is required for the biosynthesis of MK-ascarosides. We propose a model in which ACOT-15 hydrolyzes the β-keto acyl-CoA side chain of an ascaroside intermediate during β-oxidation, leading to decarboxylation and formation of the MK. Using comparative metabolomics, we identify additional ACOT-15-dependent metabolites, including an unusual piperidyl-modified ascaroside, reminiscent of the alkaloid pelletierine. The β-keto acid generated by ACOT-15 likely couples to 1-piperideine to produce the piperidyl ascaroside, which is much less dauer-inducing than the dauer pheromone, asc-C6-MK (ascr#2, 1). The bacterial food provided influences production of the piperidyl ascaroside by the worm. Our work shows how the biosynthesis of MK- and piperidyl ascarosides intersect and how bacterial food may impact chemical signaling in the worm.

Tuning of Protein Expression by Promoter Engineering.

The promoter is an essential component of an expression system since it regulates the transcriptional beginning of related genes. The optimal expression level can be achieved by employing a promoter engineering approach. Typically, creating a library of T7 promoters allows for titratable protein expression. In the process of making β-amino acid (sitagliptin intermediate) from β-keto ester, esterase from Pseudomonas stutzeri (Est PS) is used to convert the β-keto ester to β-keto acid. Subsequently, transaminase from Ilumatobacter coccineus (TAIC) transforms the β-keto acid to its corresponding β-amino acid. Here, we describe the optimization of the expression levels of Est PS for the maximum production of sitagliptin intermediate. The different promoter strengths for Est PS were built into the T7 promoters of the pET15b vector. With the help of these new co-expressing entire cells, the expressed enzyme ratio for each enzyme was determined. As the strength of the promoter of Est PS decreases, the expression level also decreases (from 100% to 10%). Conversely, the TAIC expression level is increased. This developed system produced a higher sitagliptin intermediate than enzymes' unoptimized expression level.

Reshaping the Substrate Binding Pocket of β-Amino Acid Dehydrogenase for the Synthesis of Aromatic β-Amino Acids.

By reshaping the substrate-binding pocket of β-amino acid dehydrogenase (β-AADH), some variants were obtained with up to 2560-fold enhanced activity toward the model substrates (S)-β-homophenylalanine and (R)-β-phenylalanine. A few aromatic β-amino acids were prepared with >99% ee and high isolated yields via either kinetic resolution of racemates or reductive amination of the corresponding β-keto acids. This work expands the catalytic capability of β-AADHs and highlights their practical application in the synthesis of pharmaceutically relevant β-amino acids.

Lone Pair Participation in a Decarboxylation Reaction: A New Design of a Boradecarboxylation Reaction

AbstractFor the mechanism of decarboxylation reactions, all textbooks state that electrons move from the πC=O bond. However, the most electron-donating bond orbital in the carbonyl group is expected to be the lone pair(s) on the oxygen. Thus, orbital theory with delocalization from a lone pair might be more appropriate than delocalization from the πC=O orbital. We confirmed this idea by theoretical calculations. In the transition state of 2-substituted β-keto acids, a boat conformation is expected to result in the exclusive generation of (E)-enols. Normally, the decarboxylation reaction occurs in polar solvents, so the resulting enols should be transformed into the corresponding ketones by tautomerization. The use of heteroatoms to obtain enolate or enol ethers without tautomerization would offer a diastereoselective enol(ate) synthesis with regioselectivity because the C=C double bond would always be introduced between two carbonyl groups. After screening heteroatoms by theoretical calculations, we found that boron is suitable for this purpose. We confirmed our idea by theoretical calculations, offering a new boradecarboxylation reaction for the diastereo- and regioselective production of enolates.

Organocatalytic Synthesis of Chiral Halogenated Compounds.

This account summarizes our recent efforts in the enantioselective organocatalytic synthesis of chiral halogenated compounds. The enantioselective α-halogenation of aldehydes, decarboxylative chlorination of β-keto acids, and enantioselective C-C bond formation at the trifluoromethylated prochiral carbon to yield the corresponding organohalides with chlorinated, fluorinated, or trifluoromethylated chiral stereogenic centers are discussed. We applied common organocatalysts, such as Jørgensen-Hayashi catalyst and cinchona alkaloid-derivatived catalyst, and developed novel chiral amine catalysts for these reactions. This account also discusses stereospecific derivatizations of the resulting chiral halogenated compounds via nucleophilic substitution. Thus, we synthesized many novel chiral compounds that have not been reported, even as racemates.

Stereoselective Synthesis of 1-Substituted Homotropanones, including Natural Alkaloid (−)-Adaline

The stereocontrolled synthesis of 1-substituted homotropanones, using chiral N-tert-butanesulfinyl imines as reaction intermediates, is described. The reaction of organolithium and Grignard reagents with hydroxy Weinreb amides, chemoselective N-tert-butanesulfinyl aldimine formation from keto aldehydes, decarboxylative Mannich reaction with β-keto acids of these aldimines, and organocatalyzed L-proline intramolecular Mannich cyclization are key steps of this methodology. The utility of the method was demonstrated with a synthesis of the natural product (-)-adaline, and its enantiomer, (+)-adaline.

Computational and optoelectronic quantification of semi-conducting, warm ruby red color emanating Eu3+ complexes with hetro-cyclic ligands.

Six Eu3+ complexes were synthesised with β-keto acid as main ligand and secondary ligands through liquid assisted grinding method. These complexes were characterised by various techniques such as spectroscopic technique, XRD, EDAX, SEM analysis, thermal technique, Urbach energy and optical band gap investigation. The luminous photophysical properties were studied by PL spectroscopy in solid as well as solution phase and some theoretical calculation was done to investigate the radiative (Arad) & non-radiative (Anrad) transition rate, quantum efficiency (ɸ), Judd Ofelt parameters for 5D0 → 7F2,4 transitions in both states. Judd Ofelt parameters were also calculated by the JOES software and the outcomes are well harmonised with theoretical values. The complexes have CIE color coordinate value in ruby red region and above 88.65% color purity in both phases, which made them attractive candidates for red light-emitting displays. 5D0 → 7F2 transition was proposed as a laser emission transition owing to their high branching ratio (67.18-74.24%) in solid and (60.09-74.40%) in solution phase. Computational methods were employed to determine the structure and energy of various molecular orbitals. Antimicrobial assay of complexes was also rationalised and found that the complexes are pertinent as good bactericidal and fungicidal agents in pharmaceutical industry.

Determinants of substrate specificity in a catalytically diverse family of acyl-ACP thioesterases from plants

BackgroundACYL-LIPID THIOESTERASES (ALTs) are a subclass of plastid-localized, fatty acyl-acyl carrier protein (ACP) thioesterase enzymes from plants. They belong to the single hot dog-fold protein family. ALT enzymes generate medium-chain (C6-C14) and C16 fatty acids, methylketone precursors (β-keto fatty acids), and 3-hydroxy fatty acids when expressed heterologously in E. coli. The diverse substrate chain-length and oxidation state preferences of ALTs set them apart from other plant acyl-ACP thioesterases, and ALTs show promise as metabolic engineering tools to produce high-value medium-chain fatty acids and methylketones in bacterial or plant systems. Here, we used a targeted motif-swapping approach to explore connections between ALT protein sequence and substrate specificity. Guided by comparative motif searches and computational modelling, we exchanged regions of amino acid sequence between ALT-type thioesterases from Arabidopsis thaliana, Medicago truncatula, and Zea mays to create chimeric ALT proteins.ResultsComparing the activity profiles of chimeric ALTs in E. coli to their wild-type counterparts led to the identification of interacting regions within the thioesterase domain that shape substrate specificity and enzyme activity. Notably, the presence of a 31-CQH[G/C]RH-36 motif on the central α-helix was shown to shift chain-length specificity towards 12–14 carbon chains, and to be a core determinant of substrate specificity in ALT-type thioesterases with preference for 12–14 carbon 3-hydroxyacyl- and β-ketoacyl-ACP substrates. For an ALT containing this motif to be functional, an additional 108-KXXA-111 motif and compatible sequence spanning aa77–93 of the surrounding β-sheet must also be present, demonstrating that interactions between residues in these regions of the catalytic domain are critical to thioesterase activity. The behaviour of chimeric enzymes in E. coli also indicated that aa77–93 play a significant role in dictating whether an ALT will prefer ≤10-carbon or ≥ 12-carbon acyl chain-lengths, and aa91–96 influence selectivity for substrates of fully or partially reduced oxidation states. Additionally, aa64–67 on the hot dog-fold β-sheet were shown to be important for enabling an ALT to act on 3-hydroxy fatty acyl-ACP substrates.ConclusionsBy revealing connections between thioesterase sequence and substrate specificity, this study is an advancement towards engineering recombinant ALTs with product profiles suited for specific applications.

Designing of europium(III) complexes with β-keto carboxylic acid and nitrogen-containing secondary ligand possessing optoelectrical, Urbach energy and lasing properties

Designing of europium(III) complexes with β-keto carboxylic acid and nitrogen-containing secondary ligand possessing optoelectrical, Urbach energy and lasing properties

Enzymatic C4-Epimerization of UDP-Glucuronic Acid: Precisely Steered Rotation of a Transient 4-Keto Intermediate for an Inverted Reaction without Decarboxylation.

UDP-glucuronic acid (UDP-GlcA) 4-epimerase illustrates an important problem regarding enzyme catalysis: balancing conformational flexibility with precise positioning. The enzyme coordinates the C4-oxidation of the substrate by NAD+ and rotation of a decarboxylation-prone β-keto acid intermediate in the active site, enabling stereoinverting reduction of the keto group by NADH. We reveal the elusive rotational landscape of the 4-keto intermediate. Distortion of the sugar ring into boat conformations induces torsional mobility in the enzyme's binding pocket. The rotational endpoints show that the 4-keto sugar has an undistorted 4 C1 chair conformation. The equatorially placed carboxylate group disfavors decarboxylation of the 4-keto sugar. Epimerase variants lead to decarboxylation upon removal of the binding interactions with the carboxylate group in the opposite rotational isomer of the substrate. Substitutions R185A/D convert the epimerase into UDP-xylose synthases that decarboxylate UDP-GlcA in stereospecific, configuration-retaining reactions.

Recent applications of dioxinone derivatives for macrocyclic natural product and terpenoid synthesis.

Dioxinone derivatives, a class of acetoacetate derivatives, have attracted widespread attention because of their multiple reactive sites, high reactivity, unique chemical properties, and potential synthetic applications. The dioxinone group is also stable under a wide range of reaction conditions, including strong acids, as well as a variety of transition-metal-catalysed processes, such as olefin metathesis and Pd-mediated cross-coupling. The inherent reactivity and diverse applications of dioxinones make them valuable reactive intermediates in organic synthesis. The conversion of dioxinones to acylketenes and their subsequent nucleophilic capture is also an excellent strategy for synthesising β-keto acid derivatives, which can be applied even in complex molecular synthesis. This review focuses on the recent advances in the application of dioxinones in synthetic method research and the total synthesis of natural products, highlighting the exceptional utility of these synthetic methodologies in the construction of macrocyclic cores and terpenoid skeletons. In particular, successful transformations of dioxinone fragments are discussed.

Exploration of photophysical and biological attributes of red light emitting europium (III) chelates with β-keto acid and nitrogen donor ancillary ligands

Exploration of photophysical and biological attributes of red light emitting europium (III) chelates with β-keto acid and nitrogen donor ancillary ligands

A Plurizyme with Transaminase and Hydrolase Activity Catalyzes Cascade Reactions

Engineering dual‐function single polypeptide catalysts with two abiotic or biotic catalytic entities (or combinations of both) supporting cascade reactions is becoming an important area of enzyme engineering and catalysis. Herein we present the development of a PluriZyme, TR2E2, with efficient native transaminase (k cat: 69.49±1.77 min−1) and artificial esterase (k cat: 3908–0.41 min−1) activities integrated into a single scaffold, and evaluate its utility in a cascade reaction. TR2E2 (pHopt: 8.0–9.5; T opt: 60–65 °C) efficiently converts methyl 3‐oxo‐4‐(2,4,5‐trifluorophenyl)butanoate into 3‐(R)‐amino‐4‐(2,4,5‐trifluorophenyl)butanoic acid, a crucial intermediate for the synthesis of antidiabetic drugs. The reaction proceeds through the conversion of the β‐keto ester into the β‐keto acid at the hydrolytic site and subsequently into the β‐amino acid (e.e. >99 %) at the transaminase site. The catalytic power of the TR2E2 PluriZyme was proven with a set of β‐keto esters, demonstrating the potential of such designs to address bioinspired cascade reactions.

Utilization of Judd-Ofelt theory to assess the photophysical properties of β-keto carboxylate Tb(III) complexes with heterocyclic secondary sensitizer

One binary and five ternary green glowing Tb(III) complexes with β-keto acid as primary sensitizer and heterocyclic aromatic system as secondary sensitizer, were synthesised by an eco-friendly method and characterized via various spectroscopic techniques. The assessment of photoluminescent properties in solid complexes and in solution phase with DMSO as solvent, revealed the bright green emission of complexes on irradiation of UV light due to 5D4 → 7F5 transition, whose intensity persistently increased on insertion of secondary sensitizer. Higher relative quantum yield in ternary complexes communicated the contribution of secondary sensitizers in enhancement of luminescence. Judd-Ofelt parameters (Ω2, Ω4, Ω6) were evaluated from NIR absorption spectra of complexes. Higher Ω2 value in complexes demonstrated the higher covalence character and asymmetry around Tb(III) ion. Radiative, nonradiative transition probabilities, radiative life time, branching ratios and luminescence efficiency were also estimated. Complex T6 reported to have 72.5% efficiency which signalizes superb luminescence performance of this complex. Colorimetric parameters were obtained in solid as well as solution form. Exquisite color purity and CCT value > 4000 K established these complexes as cold green light source. Band gap energy and Urbach band tail, obtained via nonlinear fitting of absorption coefficient, turned out to be in semiconductor range thus can have materiality in various photonic devices. The large value of stimulated emission cross section and narrow full width at half maxima (FWHM) of 5D4 → 7F5 emission peak suggested the prospective use of synthesised complexes as laser media for green color lasers.

Visible-Light-Promoted Cross Dehydrogenative/Decarboxylative Coupling Cascades of Glycine Ester Derivatives and β-Keto Acids.

A visible-light-induced dehydrogenative/decarboxylative coupling reaction of arylglycine derivatives and β-keto acids is described. This photocatalyst- and additive-free protocol can be applied in the efficient synthesis of γ-keto glycine derivatives under ambient conditions. Further uses of this methodology and a plausible mechanism are also demonstrated.

Achieving crimson red emission of europium (III) complexes with β-keto acids and ancillary ligands for their applications in optoelectronic devices and biomedical domain

ObjectiveIn this present work, the fabrication of europium (III) complexesdone using β-keto acid as main organic ligand and nitrogen donor chelates as ancillary ligands adopting grinding technique to investigate their applications in lighting devices, lasers, solar cells, semiconductors and bio-medical field. MethodsEuropium (III) complexes were probed employing multiple analytical and spectroscopic techniques such as EDAX mapping, elemental analysis, thermo-gravimetric analysis, ultraviolet-visible spectroscopy (UV-Vis), Nuclear magnetic resonance (NMR), infrared spectroscopy (IR) and photoluminescence (PL) studies. The biological activities such as anti-oxidant and antimicrobial assays were carried out utilizing DPPH and tube dilution method respectively. ResultsThe characteristic peak of europium (III) complexes documented at 616 nm was accountable for scarlet luminescence of these complexes. Color purity, CIE color coordinate and CCT were estimated to enable these complexes for display devices. The refractive index and energy band gap analysis portrays the applicability of these complexes in semiconductors and solar cells. Judd – Ofelt parameter shows the highly sensitive environment surrounding the europium (III) ion in complexes. The radiative parameters were determined which shows the applicability of these complexes as laser media. The quantum efficiency, decay time, branching ratio results their use in optoelectronic devices. Biological studies indicate better results for complexes than ligand in pharmaceuticals. Future prospectiveThe outcomes of investigation proclaimed the future prospective of the synthesized complexes in optoelectronics, lasers and display devices. Moreover, complexes can be probed further for biomedical purposes.

Judd-Ofelt, optical and photophysical analysis of β-ketocarboxylate Sm(III) complexes with N-donor aromatic system as secondary sensitizers

Reddish orange, Sm(III) complexes were synthesised by means of environmentally sustainable grinding method with β-keto carboxylic acid and heterocyclic aromatic systems. The photoluminescent studies revealed that 4G5/2 → 6H7/2 transition (608 nm) is responsible for the reddish orange emission of complexes when irradiated with UV light. Judd-Ofelt theory was applied to NIR absorption spectra to obtain three intensity parameters (Ω2, Ω4, Ω6) from oscillator strength. The higher Ω2 values obtained in complexes indicate greater covalence character. To assess luminescence performance, the radiative, nonradiative transition probability, radiative life time and intrinsic quantum yield were also evaluated. High quantum yield obtained for complexes signalises the superior luminescent behaviour. Exquisite color purity (89.8–96.2%) and CCT <3200 K establish these complexes as warm orange light source. The experimental data of absorption coefficient was fitted nonlinearly with Tauc's relation to obtain band gap energy. Two band gaps were reported in each complex hence the synthesised complexes can have materiality in solar cell and photovoltaic devices. The Urbach band tail width was also obtained through fitting with a new empirical relation which affirms the semiconducting properties of complexes. The investigation of antimicrobial and antioxidant properties of complexes ascertained the complexes as potent antibacterial and antioxidant agents.