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Hypothesis: Mixtures of chiral and achiral building blocks of supramolecules exhibit interesting cooperative phenomena, indicat-ed by the nonlinear composition dependence of the chiral properties. However, the nonlinear composition depend-ence of the enantioselectivity of mixed micelles is not well understood. It was hypothesized that in-depth understand-ing can be achieved by carefully investigating the composition dependence of the properties. Experiments: In this work, the nonlinear composition dependence of the enantioselectivity was found for the mixed micelle of achi-ral and chiral N-acyl amino acids by micellar electrokinetic chromatography (MEKC). Capillary electrophoresis, circu-lar dichroism (CD) spectroscopy, surface tension measurement, and fluorescence spectroscopy were used to investi-gate the mechanisms. Findings: Four mechanisms that could be causing the nonlinearity were investigated: (i) synergistic and antagonistic interac-tions of the surfactants; (ii) the chiral transfer from chiral to achiral surfactant; (iii) differences in the retention factor; and (iv) cooperative chiral recognition of the chiral and achiral surfactant. The investigation of the composition de-pendency of critical micelle concentration (CMC) and molar circular dichroism revealed that the effect of (i) and (ii) was negligibly small. The newly derived equations for (iii) and (iv) revealed that (iii) and (iv) have a major or medi-um effect on the nonlinear enantioselectivity.

Chiral pesticides account for 30% of pesticides. Pesticides are inevitably leached into the groundwater by runoff. At the watershed level, the distribution characteristics of enantiomers in sediments collected from the river network of an agricultural area near the middle and lower reaches of the Yangtze River were tested, and their potential correlations with the physicochemical properties and microbial communities of the sediments were analyzed. The sediment pollution was serious at sites 8 and 9, with their pollution source possibly being agricultural or industrial sewage. Moreover, there were higher cumulative contents of pesticide residues at sites 4, 8, and 9. Specifically, Cycloxaprid was the most detected chiral pesticide in the study area, followed by Dinotefuran and Diclofop-methyl. Additionally, Ethiprole and Difenoconazole had strong enantioselectivity in the study area. Interestingly, the enantiomers of some chiral pesticides, such as Tebuconazole, had completely different distributions at different sites. Pearson correlation analysis showed that sediment catalase and microbial biomass carbon were important factors for enantioselectivity of chiral pesticides. The effect of sediment physicochemical properties on enantioselective distribution was achieved by influencing the microorganisms in the sediment. Furthermore, the enantioselective distribution of Tebuconazole was closely related to the genus Arenimonas. Overall, the enantioselective distribution of most of the chiral pesticides was positively correlated with the prokaryotic microbial community. This study provides empirical support for agricultural non-point source pollution caused by chiral pesticides, and also lays a research foundation for exploring the factors that affect the fate of chiral pesticides in the environment.

The incorporation of poly(d-lactide) (PDLA) to form stereocomplex crystallites (SCs) within a poly(l-lactide) (PLLA) matrix is among the most effective strategies in overcoming PLLA's numerous drawbacks. However, high concentrations of PDLA (3 wt%) are required to improve PLLA's crystallization kinetics and melt strength, which is undesirable owing to PDLA's high cost. In this study, we use chain alignment as a levier to tune stereocomplex superstructure morphology to overcome these limitations. Herein, PLLA/PDLA blends were manufactured using an environmentally friendly and low-cost single step spunbond fibrillation process, yielding microfibers stretched to diameters of 5-20 μm. During this stretching process, PLLA and PDLA chains are aligned along the flow direction. SCs subsequently formed in situ upon heating, dramatically improving crystallization kinetics, melt elasticity, and tensile performance compared with neat PLLA and non-stretched blend analogues, even with low PDLA content (3 wt%). These improvements were attributed to topological variations in SC superstructures caused by alignment of PLLA and PDLA chains. The application of chain alignment in tuning SC superstructure morphology is ubiquitous in fibrillation processes.

Precise control of molecular arrangement is essential for functional molecular assemblies. A linear (I-shaped) amphiphilic block copolypeptide, polysarcosine-b-(l-Leu-Aib)

Two new neolignans and one new lignan (1-3) were obtained from the roots of Paeonia lactiflora. Their structures were unambiguously elucidated based on extensive spectroscopic analysis, single-crystal X-ray crystallography, and the calculated and experimental electronic circular dichroism (ECD) spectra. Compound 1 was a racemic mixture and successfully resolved into the anticipated enantiomers via chiral-phase HPLC. Compound 3 demonstrated moderate inhibitory activity against human carboxylesterase 2A1 (hCES2A1) with an IC

In a previous study, we identified CYP5035S7 of the white-rot fungus Polyporus arcularius with a broad activity towards monoterpenes such as p-cymene. Therefore, in this study we aimed at further exploring the substrate scope of detoxifying CYP5035S7 towards terpenes and semi-preparatively isolating some of the products via whole-cell biotransformation, in order to obtain information about the enzyme's reactivity. We noticed a clear preference for the monoterpene skeleton and elucidated a distinct regioselectivity pattern based on key structural and electronic features of its substrates. This study illustrates how minimal characterisation effort may already suffice to provide vital information on enzymatic reactivity by the comparison of structural derivatives.

The cobalt oxide-vanadium oxide (Co3O4-V2O5) combined with reduced graphene oxide (rGO) having band gap of ∼ 3.3 eV appeared as a suitable photocatalyst for selective oxidation of 2-naphthol to BINOL. C2-symmetric BINOL was achieved with good yield using hydrogen peroxide as the oxidant under UV-light irradiation. The same catalyst was chirally modified with cinchonidine and a newly synthesized chiral Schiff base ligand having a sigma-hole center. The strong interaction of the chiral modifiers with the cobalt-vanadium oxide was truly evident from various spectroscopic studies and DFT calculations. The chirally modified mixed metal oxide transformed the oxidative C C coupling reaction with high enantioselectivity. High enantiomeric excess upto 92 % of R-BINOL was obtained in acetonitrile solvent and hydrogen peroxide as the oxidant. A significant achievement was the formation of S-BINOL in the case of the cinchonidine modified catalyst and R-BINOL with the Schiff base ligand anchored chiral catalyst. The UV-light induced catalytic reaction was found to involve hydroxyl radical as the active reactive species. The spin trapping ESR and fluorescence experiment provided relevant evidence for the formation of such species through photodecomposition of hydrogen peroxide on the catalyst surface. The chiral induction to the resultant product was found to induce through supramolecular interaction like O H…π, H…Br interaction. The presence of sigma hole center was believed to play significant role in naphtholate ion recognition during the catalytic cycle.

4,8-Sphingadienines (SD), metabolites of glucosylceramides (GlcCer), are sometimes determined as key mediators of the biological activity of dietary GlcCer, and cis/trans geometries of 4,8-SD have been reported to affect its activity. Since regulating excessive activation of mast cells seems an important way to ameliorate allergic diseases, this study was focused on cis/trans stereoisomeric-dependent inhibitory effects of 4,8-SD on mast cell activation. Degranulation of RBL-2H3 was inhibited by treatment of 4-cis-8-trans- and 4-cis-8-cis-SD, and their intradermal administrations ameliorated ear edema in passive cutaneous anaphylaxis reaction, but 4-trans-8-trans- and 4-trans-8-cis-SD did not. Although the activation of mast cells depends on the bound IgE contents, those stereoisomers did not affect IgE contents on RBL-2H3 cells after the sensitization of anti-TNP IgE. These results indicated that 4-cis-8-trans- and 4-cis-8-cis-SD directly inhibit the activation of mast cells. In conclusion, it was assumed that 4,8-SD stereoisomers with cis double bond at C4-position shows anti-allergic activity by inhibiting downstream pathway after activation by the binding of IgE to mast cells.

Naturally occurring DNA, RNA, and proteins predominantly exist in only one enantiomeric form (homochirality). Advances in biotechnology and chemical synthesis allow the production of the respective alternate enantiomeric form, enabling access to mirror-image versions of these natural biopolymers. Exploiting the unique properties of such mirror molecules has already led to many applications, such as biostable and nonimmunogenic therapeutics or sensors. However, a 'roadblock' for unlocking the mirror world is the lack of biological systems capable of synthesizing critical building blocks including mirror oligonucleotides and oligopeptides to reducing cost and improve purity. Here, we provide an overview of the current progress, applications, and challenges of the molecular mirror world by identifying milestones towards mirroring life.

Drug isomers may differ in their proarrhythmia risk. An interesting example is the drug sotalol, an antiarrhythmic drug comprising d- and l- enantiomers that both block the hERG cardiac potassium channel and confer differing degrees of proarrhythmic risk. We developed a multi-scale in silico pipeline focusing on hERG channel - drug interactions and used it to probe and predict the mechanisms of pro-arrhythmia risks of the two enantiomers of sotalol. Molecular dynamics (MD) simulations predicted comparable hERG channel binding affinities for d- and l-sotalol, which were validated with electrophysiology experiments. MD derived thermodynamic and kinetic parameters were used to build multi-scale functional computational models of cardiac electrophysiology at the cell and tissue scales. Functional models were used to predict inactivated state binding affinities to recapitulate electrocardiogram (ECG) QT interval prolongation observed in clinical data. Our study demonstrates how modeling and simulation can be applied to predict drug effects from the atom to the rhythm for dl-sotalol and also increased proarrhythmia proclivity of d- vs. l-sotalol when accounting for stereospecific beta-adrenergic receptor blocking.

Background Ketamine has cardiac excitatory side-effects. Currently, data on the effects of ketamine and metabolite concentrations on cardiac output are scarce. We therefore developed a pharmacodynamic model derived from data from a randomised clinical trial. The current study is part of a larger clinical study evaluating the potential mitigating effect of sodium nitroprusside on the psychedelic effects of ketamine. Methods Twenty healthy male subjects received escalating esketamine and racemic ketamine doses in combination with either placebo or sodium nitroprusside on four visits: (i) esketamine and placebo, (ii) esketamine and sodium nitroprusside, (iii) racemic ketamine and placebo, and (iv) racemic ketamine and sodium nitroprusside. During each visit, arterial blood samples were obtained and cardiac output was measured. Nonlinear mixed-effect modelling was used to analyse the cardiac output time-series data. Ketamine metabolites were added to the model in a sequential manner to evaluate the effects of metabolites. Results A model including an S-ketamine and S-norketamine effect best described the data. Ketamine increased cardiac output, whereas modelling revealed that S-norketamine decreased cardiac output. No significant effects were detected for R-ketamine, metabolites other than S-norketamine, or sodium nitroprusside on cardiac output. Conclusions S-Ketamine, but not R-ketamine, increased cardiac output in a dose-dependent manner. In contrast to S-ketamine, its metabolite S-norketamine reduced cardiac excitation in a dose-dependent manner. Clinical trial registration Dutch Cochrane Center 5359.

Curcumin is a potential candidate in cancer therapy due to its ability to inhibit many signalling pathways at the same time of exposure because of its unique content of aromatic ring, B diketone, olefinic linker, and O methoxy phenolic groups. Its applications in biomedical therapy is limited because of its sensitivity, and its rapid degradation. In the current study, curcumin inserted into polyelectrolyte pairs (protamine and dextran) and then was functionalized by folic acid conjugated chitosan used for the first time, as theranostic system. Such this strategy allows to improve its mucoadhesion and penetration that increases their accumulation inside cancer cells. CUR-LbL NPs were then used to investigate drug release inside Human Mammary Carcinoma (MCF-7 cell lines) after their incubations for 3 h, 6 h and 24 h. Flow cytometry indicated that the percentages of apoptosis, necrosis and cell cycle arrest were increased significantly in MCF-7 cell lines treated by CUR-LbL NPs. Furthermore, SEM image showed many debris in the section of MCF-7 treated by CUR-LbL NPs. Here, it can be summarized that curcumin functionalized by multi-layered polyelectrolyte capsules can be used as a model to study the fate of the adsorbed nanocarriers and to investigate the drug release inside cells.

Metal organic frameworks (MOFs) are hybrid organic inorganic materials with unique properties such as well-defined pore structure, extremely high surface area, excellent chemical-thermal stability. MOFs-based constructs have been extensively engineered and used for applications, such as enzyme immobilization for bio-catalysis. To obtained a zeolitic imidazole framework-8 (ZIF-8) for enzyme immobilization, Candida rugosa lipase (CRL) was pretreated with calix [4]arene tetracarboxylic acid (Calix) and reacted with Zn and imidazole by co-precipitation method. The prepared biocomposite was characterized by SEM, EDX, FT-IR, and XRD. The prepared CRL@Calix-ZIF-8 with high encapsulation efficiency showed improved resistance to alkali and thermal conditions. The CRL@Calix-ZIF-8 with the biocatalytic activity was 2-folds higher than that of the CRL@ZIF-8 (without Calix). The free lipase lost its catalytic activity completely at 60 °C after 100 min, while the CRL@Calix-ZIF-8 and CRL@ZIF-8 retained about 84% and 73%. It was found that CRL@Calix-ZIF-8 and CRL@ZIF-8 still retained ~83 and 67% of catalytic activity after its 6th use, respectively. The kinetic resolution of the immobilized lipases was examined for enantioselective hydrolysis of racemic naproxen methyl ester. CRL@Calix-ZIF-8 showed enantioselectivity against the racemic naproxen methyl ester, with E = 183 and 131 compared to the CRL@ZIF-8.

Lipase immobilization with hydrophobic interaction is of interesting exploration, and some functionalized groups on supports are special for activity increasing. To achieved a good performance of cost-effective immobilization on macro-supports for feasible usage and recycle, eco-friendly PLA-based 3D printing macro-scaffolds with fabrication was designed, and phenyl groups with different length of linkers and combined two kinds of groups were anchored for lipase YCJ01 binding with improving payload, the highest enzyme expression of 2227.5 U/g, activity recovery of 137.3%, and increasing specific activity of 815.9 U/mg were attained by using PLA@AMTS-C7-Ph/PLA@AMTS-C9-Ph scaffolds as carries. The immobilized lipase YCJ01 on bifunctionalized 3D printing scaffolds was further applied to the efficient resolution of racemic 1-indanol (267 mM) with high stereoselectivity using a binary solvent system. The immobilized lipase YCJ01 could control the over transesterification of (S)-1-indanol and exhibit good operational stability of repetitive usage for 9 cycles. This is beneficial to obtain the high enantiomerical pure product by feasible separation of immobilized biocatalyst without rigorous operation.

In this study, Graphene Oxide (GO) was used to screen the binding with the aptamers of L-carnitine chiral enantiomers. The ssDNA library was prepared by the method of Lambda exonuclease. In addition, a simple casing device was designed to improve the purification and recovery efficiency of the small ssDNA fragments in the process of screening. Finally, more than 160,000 aptamer sequences were obtained by high-throughput sequencing. We determined the strongest affinity aptamer sequence, CA04, by the Resonance Rayleigh scattering (RRS) technology. We also analyzed the key binding sites (in the 16th position case) of the truncated aptamer sequence CAD10. Interestingly, we found that aptamer CA10 and CA06 were both C-rich bases through sequence alignment and analysis, and the aptamer CA10 was confirmed that the CA10 and CA06 were formed under acidic conditions (pH 4.5) by CD spectrum and ESI-MS analysis. The interaction between gold nanoparticles (AuNPs) and functionalized aptamer CA10 was analyzed. We used Site-directed mutagenesis design and QGRS Mapper to optimize aptamer CA10, where an optimal aptamer CA10-03 were obtained after affinity analysis. It is also proved to be an effective method to obtain stronger affinity aptamer. Meanwhile, Native-PAGE and UV spectrum analysis were performed on the mutation sequences, and the interaction with ThT was analyzed. Finally, it is hoped that my study can provide help for later identification and detection of L-carnitine.

Only a few known epoxide hydrolases (EHs) displayed activity towards o-nitrostyrene oxide (4a), presumably owing to the large steric hindrance caused by o-nitro substituent. Therefore, excavating EHs with high activity and enantio- and/or regio-selectivity towards racemic (rac-) 4a is essential but challenging. Here, AuEH2 from Aspergillus usamii was expressed in E. coli BL21(DE3). E. coli/Aueh2, an E. coli transformant expressing AuEH2, possessed EH activities of 16.2–184 U/g wet cell towards rac-styrene oxide (1a) and its derivatives (2a–13a), and the largest enantiomeric ratio of 96 towards rac-4a. The regioselectivity coefficients, βR and βS, of AuEH2 were determined to be 99.2% and 98.9%, suggesting that it regiopreferentially attacks the Cβ in the oxirane rings of (R)- and (S)-4a. Then, the nearly perfect kinetic resolution of 20 mM rac-4a in pure water was carried out using 20 mg/mL wet cells of E. coli/Aueh2 at 25 °C for 50 min, retaining (S)-4a with over 99% ees and 48.9% yields, while producing (R)-o-nitrophenyl-1,2-ethanediol (4b) with 95.3% eep and 49.8% yieldp. To elucidate the molecular mechanism of AuEH2 with high enantiopreference for (R)-4a, its crystal structure was solved by X-ray diffraction and the molecular docking of AuEH2 with (R)- or (S)-4a was simulated.

Introduction In vivo imaging methods such as Positron Emission Tomography (PET) can be used to examine the relationship between in vitro binding affinity and in vivo occupancy of binding sites in the brain for new drug candidates. In this study, PET imaging in monkey brain was used to evaluate that correlation for a set of four diastereomers of the compound dihydrotetrabenazine (DTBZ), the pharmacologically active metabolite of the drug tetrabenazine. Methods PET studies of DTBZ diastereomers were completed in a single monkey brain. In vivo occupancies (ED50) were estimated using multiple drug doses and the vesicular monoamine transporter 2 specific radioligand (+)-α-[11C] DTBZ, employing a test-retest sequence of control PET scan, drug administration and a second PET scan completed on a single day. Results DTBZ has three chiral carbon centers and eight possible stereoisomers, and in vivo occupancy of the target site VMAT2 was observed only for the four diastereomers of DTBZ having the 11bR absolute configuration. The estimated in vivo occupancies (ED50 values from 0.023 to >3.15 mg/kg) correlated well (R2 = 0.95) with the in vitro binding affinities (Ki values of 4 to 600 nM for the VMAT2), and an even better correlation (R2 = 0.99) was found for the three isomers with in vitro binding affinities Conclusions If the physiochemical (MW, log P, pKa) or physiological (metabolism, transport, protein binding) properties of a set of drug stereoisomers are considered similar, the binding affinities determined from in vitro assays may predict the in vivo occupancies of the target binding site in the monkey brain.

This work reports on targeted UHPLC-tandem mass spectrometry methods for the chiral separation of anteiso-methyl branched fatty acids (aiFAs). The methods involve precolumn derivatization with 1-naphthylamine and chiral separation on Chiralpak IG-U. anteiso-Methyl branched fatty acids with up to eight carbons can be separated. A method was used for the assignment of the absolute configuration of an aiFA present as fatty acyl residue of the teicoplanin mixture, namely teicoplanin RS3. Furthermore, the excellent methylene selectivity and improved selectivity for constitutional isomers of the polysaccharide columns was exploited for the elucidation and structural confirmation of previously unknown fatty acyl residues in teicoplanin. This shows the versatility and practical applicability of polysaccharide columns as orthogonal stationary phases to reversed-phase for structural elucidation of natural compounds. The developed methods are useful tools for related subdisciplines such as targeted metabolomics and lipidomics.

Environmental risks associated with neonicotinoid insecticides have attracted considerable attention. This study systematically investigated the stereoselective behavior of dinotefuran in a water-sediment system. The results showed that S-dinotefuran accumulated more easily in sediment and zebrafish. Although dinotefuran enantiomers and metabolites present a low risk to aquatic organisms, the risk of dinotefuran enantiomers to sediment organisms should be considered. Additionally, S-dinotefuran induced more remarkable oxidative damage in zebrafish than that of R-dinotefuran. Nevertheless, R-dinotefuran remarkably activated antioxidant and detoxifying enzymes. Multi-omics analyses revealed that S-dinotefuran induced more differentially expressed genes (DEGs) and differentially expressed proteins (DEPs) in zebrafish. In particular, S-dinotefuran inhibited the expression of ribosome- and proteasome-related genes and proteins, affecting the synthesis and degradation of proteins in zebrafish. R-dinotefuran remarkably activated peroxisome-related genes and proteins, thereby enhancing antioxidant and detoxification abilities of zebrafish. The stereoselective interactions between dinotefuran enantiomers and key DEPs were elucidated using AlphaFold2 modeling and molecular docking techniques, which may serve as the main reason for stereoselective subchronic toxicity. The present study is beneficial for the correct use of dinotefuran and provides an effective means for elucidating the mechanism of the stereoselective behavior of chiral compounds.

As a special category of pesticides, chiral pesticides have increased the difficulty in investigating pesticide toxicity. Based on their usage, chiral pesticides can be divided into insecticides, herbicides, and fungicides. Over the past decades, great efforts have been made on elucidating their toxicological effects. However, no literature has reviewed the enantioselective toxicity of chiral pesticides since 2014. In recent years, more chiral pesticides have been registered for application. As such, huge research progresses have been achieved in enantioselective toxicity of chiral pesticides. Generally, more researches have remedied the knowledge gap in toxicological effects of old and new chiral pesticides. And the toxicological endpoints being evaluated have become more specific rather than centering on basic toxicity and target organisms. Besides, the underlying mechanisms accounting for the enantioselectivity in toxicological effects of chiral pesticides have been discussed as well. All in all, this review provides the critical knowledge for risk assessments, and help to drive the green-technology of single- or enriched-enantiomer pesticides and formulation of relevant laws and regulations.

Herein, we describe the impact of the introduction of tertiary amines as additive during the enzymatic kinetic resolution via deacylation of some arylalkyl acetates 1a-8a under promiscuous conditions. Two CAL-B preparations were examined: Novozym®435 and CHIRAZYME® L-2, c.-f. C2, lyo. The influence of the introduction of seven amines is checked: Triethylamine (Et

Chiral analysis of food components can provide important information for food quality, bioactivity and safety. Determination of enantiomeric ratios in food is a tedious task, due to the poor resolution and insufficient sensitivity for simultaneous discrimination and quantification of trace amounts of d-form metabolites. Herein, a high-throughput, high-sensitive and high-resolution method was developed for simultaneously determining enantiomeric ratios of multiple chiral α-hydroxy/amino acids (HA/AAs) from fermented milks in one-run by [d

Lipoxygenases (LOXs) are implicated in the biosynthesis of pro- and anti-inflammatory lipid mediators involved in immune cell signaling, most of which catalyze peroxidation of polyunsaturated fatty acids by distinct regio- and stereoselectivity. Current reports suggested that conserved amino acid, Gly in R-LOXs and Ala in S-LOXs, in the catalytic domain play an important role in determining the position as well as the stereochemistry of the functional group. Recently, we have confirmed that the catalytic specificity of cyanobacterial lipoxygenase, named Osc-LOX, with alanine at 296 was 13S-type toward linoleic acid, and producing a 17S- hydroxy-docosahexaenoic acid from docosahexaenoic acid (DHA). Here, we aimed to change the catalytic property of LOX from13S-LOX to 9R-LOX by replacing Ala with Gly and to produce a lipid mediators different from the wild-type using DHA. Finally, we succeeded in generating human endogenous a 13R-hydroxy-docosahexaenoic acid and a 13R,20-dihydroxy-docosahexaenoic acid from DHA through an enzymatic reaction using the Osc-LOX-A296G. Our study could enable physiological studies and pharmaceutical research for the 13R,20-dihydroxy-docosahexaenoic acid.

Epoxide hydrolases (EHs) have been characterized and engineered as biocatalysts that convert epoxides to valuable chiral vicinal diol precursors of drugs and bioactive compounds. Nonetheless, the regioselectivity control of the epoxide ring opening by EHs remains challenging. Alp1U is an α/β-fold EH that exhibits poor regioselectivity in the epoxide hydrolysis of fluostatin C (compound 1) and produces a pair of stereoisomers. Herein, we established the absolute configuration of the two stereoisomeric products and determined the crystal structure of Alp1U. A Trp-186/Trp-187/Tyr-247 oxirane oxygen hole was identified in Alp1U that replaced the canonical Tyr/Tyr pair in α/β-EHs. Mutation of residues in the atypical oxirane oxygen hole of Alp1U improved the regioselectivity for epoxide hydrolysis on 1. The single site Y247F mutation led to highly regioselective (98%) attack at C-3 of 1, whereas the double mutation W187F/Y247F resulted in regioselective (94%) nucleophilic attack at C-2. Furthermore, single-crystal X-ray structures of the two regioselective Alp1U variants in complex with 1 were determined. These findings allowed insights into the reaction details of Alp1U and provided a new approach for engineering regioselective epoxide hydrolases.

Central administration of L-arginine was reported to attenuate stress responses in neonatal chicks. The present study aimed to elucidate the differential effects of centrally administered L-arginine and its enantiomer, D-arginine, on the stress response in chicks and the associated mechanisms. Intracerebroventricular injection of L-arginine attenuated acute isolation stress by inducing sleep-like behavior, while central administration of D-arginine potentiated the stress response, reducing the time spent standing motionless with eyes open and increasing distress vocalizations compared to the control. The brain concentrations of amino acids and monoamines following L- and D-arginine administration during stress were also determined. L-Arginine significantly increased the mesencephalic L-glutamine concentration. D-Arginine administration did not affect the levels of L-arginine or other amino acids in the examined brain regions. 3,4-Dihydroxyphenylacetic acid (DOPAC) level and dopamine (DA) metabolic rate (DOPAC/DA) were significantly higher in the diencephalon in the D-arginine group compared to the L-arginine group, while the mesencephalic DA level was significantly lower in the D-arginine group compared to the control. In vitro experiment using the brain slice culture demonstrated that extracellular perfusion of D-arginine significantly elevated the mRNA expression level of monoamine oxidase B, the major enzyme involved in DA metabolism, in the locus coeruleus region of the brainstem. In conclusion, in neonatal chicks, central administration of D-arginine exerted a stimulant effect on the stress response, in contrast to the stress-attenuating effects of L-arginine, partly through an effect on brain dopaminergic metabolism and not through competition with the L-stereoisomer.

Cyanobacteria can grow photoautotrophically, producing a range of substances by absorbing sunlight and utilizing carbon dioxide, and can potentially be used as industrial microbes that have minimal sugar requirements. To evaluate this potential, we explored the possibility of l -glutamate production using the Synechocystis sp. PCC6803. The ybjL gene encoding the putative l -glutamate exporter from Escherichia coli was introduced, and l -glutamate production reached 2.3 g/L in 143 h (34°C, 100 μmol m−2 s−1). Then, we attempted to produce two flavor substances, (S)-linalool, a monoterpene alcohol, and the sesquiterpene (+)-valencene. The Synechocystis sp. PCC6803 strain in which the linalool synthase gene (LINS) from Actinidia arguta (AaLINS) was expressed under control of the tac promoter (GT0846K-Ptac-AaLINS) produced 11.4 mg/L (S)-linalool in 160 h (30°C, 50 μmol m−2 s−1). The strain in which AaLINS2 and the mutated farnesyl diphosphate synthase gene ispA∗ (S80F) from E. coli (GT0846K-PpsbA2-AaLINS-ispA∗) were expressed from the PpsbA2 promoter accumulated 11.6 mg/L (S)-linalool in 160 h. Genome analysis revealed that both strains had mutations in slr1270, suggesting that loss of Slr1270 function was necessary for high linalool accumulation. For sesquiterpene production, the valencene synthase gene from Callitropsis nootkatensis and the fernesyl diphosphate synthase (ispA) gene from E. coli were introduced, and the resultant strain produced 9.6 mg/L of (+)-valencene in 166 h (30°C, 50 μmol m−2 s−1). This study highlights the production efficiency of engineered cyanobacteria, providing insight into potential industrial applications.

The enzyme β-carotene oxygenase 2 (BCO2) converts carotenoids into more polar metabolites. Studies in mammals, fish, and birds revealed that BCO2 controls carotenoid homeostasis and is involved in the pathway for vitamin A production. However, it is controversial whether BCO2 function is conserved in humans, because of a 4-amino acid long insertion caused by a splice acceptor site polymorphism. We here show that human BCO2 splice variants, BCO2a and BCO2b, are expressed as pre-proteins with mitochondrial targeting sequence (MTS). The MTS of BCO2a directed a green fluorescent reporter protein to the mitochondria when expressed in ARPE-19 cells. Removal of the MTS increased solubility of BCO2a when expressed in Escherichia coli and rendered the recombinant protein enzymatically active. The expression of the enzymatically active recombinant human BCO2a was further improved by codon optimization and its fusion with maltose-binding protein. Introduction of the 4-amino acid insertion into mouse Bco2 did not impede the chimeric enzyme's catalytic proficiency. We further showed that the chimeric BCO2 displayed broad substrate specificity and converted carotenoids into two ionones and a central C14-apocarotendial by oxidative cleavage reactions at C9,C10 and C9',C10'. Thus, our study demonstrates that human BCO2 is a catalytically competent enzyme. Consequently, information on BCO2 becomes broadly applicable in human biology with important implications for the physiology of the eyes and other tissues.

Carbohydrates play important roles in life science, but their synthesis is always hampered by their complicated chemical structures. Scientists have never stopped trying to solve the problem of glycan synthesis from various aspects. Here a brief overview of recent progress in glycan synthesis, including chemical approaches, chemoenzymatic approaches, and automated synthesis, will be discussed, focusing on the efficiency of new glycosylation methods, the stereoselectivity of coupled products, and their applications in the assembly of complex glycan chains.

Starting from cyclooctatetraene, bis-homoconduritols with cis-inositol and allo-inositol (or bicyclo[4.2.0]octane motif) structures were synthesized. Photooxygenation of trans-7,8-dibromo-bicyclo[4.2.0]octa-2,4-diene allowed the preparation of tricyclic endoperoxide. The compound diacetate was obtained by reduction of endoperoxide with thiourea followed by acetylation reaction. Removal of halides with zinc dust in acetic acid yielded the dien-diacetate, a key compound of the designed molecules. OsO

Development of chiral drugs has become one of the most important subjects for academic research and pharmaceutical industry. Even though it has been well known that a pair of enantiomers of a chiral drug frequently have different biological and physiological effects, a certain number of chiral drugs are currently still commercialized and used in the form of racemate. In the above context, investigation of enantio-differentiating release and enantioselective effects may lead to alternative use approaches for chiral drugs. In spite of the noticeable advancements, there has been no review paper yet focused on enantioselective release, except the earlier one published by us (Ind. Eng. Chem. Res. 2016, 55, 6037). The present paper reviews the latest advancements made in enantioselective release, with emphasis on the construction of chiral releasing materials. The currently existing difficulty in performing enantioselective release and the emerging issues associated with chiral drugs (and chiral agrochemicals, etc.) are summarized. The issues include: enantioselective effects in chiral carrier-chiral guest compound and chiral carrier-achiral guest compound releasing systems; chiral inversion in release process; chiral pollutants in environment, etc. The review paper might provide new insights into the development and use of chiral drugs.

The pressure-temperature phase diagram of the dimorphism of the contraceptive drug gestodene is constructed using the temperature and enthalpy of fusion of form I (469.5K, 107Jg-1), and those of the endothermic transition from form II to form I (311K, 8.52Jg-1). At ordinary pressure, the sign of the enthalpy of this transition indicates that these polymorphs are enantiotropically related and that form II, whose melting temperature is calculated to be about 452K, is the stable form at room temperature. Considering the inequality in the specific volumes of the two polymorphs, it is shown that the two forms remain enantiotropically related on increasing pressure, because the I-II equilibrium and the melting equilibria I-L and II-L diverge as a consequence of the negative slope dP/dT of the solid-solid equilibrium. In addition, it is demonstrated that the heats of dissolution, inferred from solubility measurements, lead to virtually the same value of the heat of transition from II to I as for the differential scanning calorimetry measurements.

Spiropyrans are a class of photochromic compounds that feature two main isoforms, a closed spiropyran (SP) and an open merocyanine (MC), which are inter-switchable with UV and visible light. The double bond containing MC isomer has several resonance forms with different optical spectra. Consequently, the overall spectral characteristic of the spiropyran solution depends on the relative contribution of each resonance form. In addition, the reversible changes of the absorbance or fluorescence spectra of MC are dictated by the environment. By utilizing these properties, spiropyran and its derivatives have been employed to monitor a variety of molecules and ions that can alter the resonance forms of MC. In this review, we provide a comprehensive discussion on a range of species that can be detected by spiropyran including metal ions, anions, acids, solvents, and even biomolecules, the major analytical detection mechanisms involved, and the novelty of such techniques. This study establishes spiropyran as a versatile platform for the detection of both organic and inorganic target molecules.

Exploitation of enzymes in biocatalytic processes provides scope both in the synthesis and degradation of molecules. Enzymes have power not only in their catalytic efficiency, but their chemoselectivity, regioselectivity, and stereoselectivity means the reactions they catalyze are precise and reproducible. Focusing on carbohydrate processing enzymes, this review covers advances in biocatalysis involving carbohydrates over the last 2-3 years. Given the notorious difficulties in the chemical synthesis of carbohydrates, the use of enzymes for synthesis has potential for significant impact in the future. The use of catabolic enzymes in the degradation of biomass, which can be exploited in the production of biofuels to provide a sustainable and greener source of energy, and the synthesis of molecules that have a range of applications including in the pharmaceutical and food industries will be explored.

Pictet-Spenglerases provide a key role in the biosynthesis of many biologically active alkaloids. There is increasing use of these biocatalysts as an alternative to traditional organic synthetic methods as they provide stereoselective and regioselective control under mild conditions. Products from these enzymes also contain privileged drug scaffolds (such as tetrahydroisoquinoline or β-carboline moieties), so there is interest in the characterization and use of these enzymes as versatile biocatalysts to synthesize analogs of the corresponding natural products for drug discovery. This review discusses all known Pictet-Spenglerase enzymes and their applications as biocatalysts.

Chirality seems to play a key role in mineralization. Indeed, in biominerals, the biomolecules that guide the formation and organization of inorganic crystals and help construct materials with exceptional mechanical properties, are homochiral. Here, we show that addition of homochiral l -(+)-tartaric acid improved the mechanical properties of brushite bioceramics by decreasing their crystal size, following the classic Hall-Petch strengthening effect; d -(-)-tartaric acid had the opposite effect. Adding l -(+)-Tar increased both the compressive strength (26 MPa) and the fracture toughness (0.3 MPa m1/2) of brushite bioceramics, by 33% and 62%, respectively, compared to brushite bioceramics without additives. In addition, l -(+)-tartaric acid enabled the fabrication of cements with high powder-to-liquid ratios, reaching a compressive strength and fracture toughness as high as 32.2 MPa and 0.6 MPa m1/2, respectively, approximately 62% and 268% higher than that of brushite bioceramics prepared without additives, respectively. Characterization of brushite crystals from the macro- to the atomic-level revealed that this regulation is attributable to a stereochemical matching between l -(+)-tartaric acid and the chiral steps of brushite crystals, which results in inhibition of brushite crystallization. These findings provide insight into understanding the role of chirality in mineralization, and how to control the crystallographic structure of bioceramics to achieve high-performance mechanical properties. Statement of Significance Calcium-phosphate cements are promising bone repair materials. However, their suboptimal mechanical properties limit their clinical use. Natural biominerals have remarkable mechanical properties that are the result of controlled size, shape and organization of their inorganic crystals. This is achieved by biomineralization proteins that are homochiral, composed of l - amino acids. Despite the importance of chiral l -biomolecules in biominerals, using homochiral molecules to fabricate bone cements has not been studied yet. In this study, we showed that homochiral l -(+)-tartaric acid can regulate the crystal structure and improve the mechanical properties of a calcium-phosphate cement. Hence, these findings open the door for a new way of designing strong bone cement and highlight the importance of chirality in bioceramics.

The integration of biocatalysis with chemocatalysis combines the excellent selectivity of the former with the robust reactivity of the latter and offers many advantages, such as lower cost, higher yield, enhanced selectivity, as well as less waste generation. In spite of the challenge of incompatibilities between different classes of catalysts, recent advances in synthetic chemistry and biology provide ample opportunities for multistep cascade transformations that combine biocatalysis and chemocatalysis. Herein, we review recent progress in merging biocatalysis with chemocatalysis, highlighting selected examples of photo-/electricity-driven biotransformations and recently developed strategies for addressing the catalyst incompatibility issue.

An open reading frame of sleh1, a gene encoding for a novel epoxide hydrolase from Solanum lycopersicum (SlEH1), was amplified by RT-PCR and expressed in E. coli BL21(DE3). The substrate spectrum assay showed that E. coli/sleh1 had EH activities towards all tested substrates except for racemic (rac-) 5a, and the highest enantiomeric ratio (E > 200) towards rac-2a, retaining (R)-2a with 99.1% ees and 49.2% yields and affording (R)-2b with 89.8% eep and 46.7% yieldp. Besides, E. coli/sleh1 also hydrolyzed of rac-7a–9a with moderate regioselectivities, producing (S)- or (R)-7b–9b with 40.5–51.3% eep and 69.4–75.2% yieldp. The pH optimum and stability of the purified SlEH1 were 7.5 and at a range of 6.5–8.5, and it was thermostable at or below 40 °C. Its catalytic efficiency (kcatS/KmS = 7.49 mM−1 s−1) for (S)-2a was much higher than that for (R)-2a. The gram-scale kinetic resolution of 150 mM rac-2a was carried out by E. coli/sleh1 at 20 °C for 8 h, producing (R)-2a with 98.2% ees and 45.3% overall yields after purification by silica gel column chromatography. Furthermore, the source of extremely high enantioselectivity of SlEH1 towards rac-2a was analyzed by molecular docking simulations.

Cytochrome P450 enzymes (P450 or CYP) are some of the most versatile biocatalysts, and offer advantages for oxidizing unreactive C–H bonds in mild conditions. In this study, we identified a novel cytochrome P450 154C2 from Streptomyces avermitilis and characterized its function in 2α-hydroxylation of testosterone with regio- and stereoselectivity. To investigate the efficiency of electron transfer, we conducted biotransformation using two different P450 redox partners—RhFRED (RhF reductase domain) from Rhodococcus sp. and Pdx (putidaredoxin)/Pdr (putidaredoxin reductase) from Pseudomonas putida and revealed that RhFRED was more effective than Pdx/Pdr, especially in vivo. The Km and kcat values for testosterone were estimated to be 0.16 ± 0.05 mM and 0.13 ± 0.02 min−1, and kcat/Km was 0.81 min−1 mM−1. We also determined the crystal structure of the substrate-free form of CYP154C2 at 1.5 A resolution. The structure has a closed conformation, and the substrate binding pocket is narrow, which can explain the strict substrate specificity of the enzyme.

A polyaminosaccharide (chitosan, CS) and an aminosaccharide ( d -galactosamine, GalN) were integrated together via hydrothermal assembly to obtain a bis-aminosaccharides composite (CS-GalN), and a novel and facile chiral sensing platform based on CS-GalN modified glassy carbon electrode (CS-GalN/GCE) was fabricated and used for electrochemical recognition of tyrosine (Tyr) enantiomers. CS-GalN composite was characterized by Fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS) and contact angle goniometry. It was observed that CS-GalN composite exhibited different binding ability for Tyr enantiomers. Under optimized experimental conditions, the oxidation peak current ratio of L-Tyr to D-Tyr (IL/ID) and the difference between their peak potentials (ΔEp = ED-EL) were 1.70 and 28 mV at CS-GalN/GCE by square wave voltammetry (SWV). In addition, the peak currents increase linearly with the concentration of Tyr enantiomers in the concentration range 0.01–1.00 mM with detection limits of 0.65 μM and 0.86 μM for L-Tyr and D-Tyr (S/N = 3), respectively. CS-GalN/GCE also exhibited the ability to determine the percentage of D-Tyr in the racemic mixture. In addition, CS-GalN/GCE possessed remarkable sensitivity, great stability as well as fine reproducibility. It could be concluded that the chiral interface of CS-GalN/GCE can provide an ideal platform for electrochemical recognition and determination of Tyr enantiomers.

Using over fifty steroidal ligands, CYP3A4 Template system established in our previous study (DMPK 34: 113-125, 2019) has been evaluated for the applicability for prediction of regioselective metabolisms of steroids in the present study. Plural regional interactions near Site of Oxidation of CYP3A4 (Slide-down and Adaptation) are newly defined for steroid ligands in addition to previously characterized Trigger- and IJL-interactions on Template. Interaction of steroids at ring-A with CYP3A4 residue (Front-residue), at the facial side of Ring B of Template, determined the availability of ligand sitting at Rings A and B of Template. Steroids having 3-one-4-ene structures, which are not stacked on Front-residue, thus slide down for their 6-oxidations. Some steroids with 3β-ol structures undergo the further right-side movement (Adaptation) for their 7-oxidations. Similar overpassing phenomena are also expected for steroid 15/16-oxidations and 2/1-oxidations. Allowable width on ligand accommodation was also defined as Width-gauge of Template. Reciprocal comparison of sittings of steroids on Template with experimental data offered idea of CYP3A4-mediated oxidations of steroids through seven distinct types of placements on Template and of the relationship with their usage abundance. The present system would offer practical way for structural identification and verification of CYP3A4-mediated metabolisms of various types of steroids.

Although d-amino acids are less prevalent in nature, they have been detected in mammals (including humans) and it is widely accepted that they might play important physiological roles. While an analytical method for chiral amino acid profiling is strongly required, it has not been well-established because of the difficulties associated with analysis. A high-sensitivity and high-throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) analytical method was recently reported by our group for chiral amino acids; however, it lacked sufficient repeatability for several d-amino acids. Thus, the aim of this research was to reduce the experimental variation of chiral amino acid analysis. By installing an automatic switching valve system in LC-MS/MS, it was possible to reduce the relative standard deviations of d-amino acid ratios (d/(d+l)) in rat urine obtained from three technical replicates. The results indicated that the automatic switching valve system was effective in minimizing the variation of d-amino acid ratios, and could be applied for profiling d-amino acids because of its high repeatability.

An intracellular esterase (BCE) from Bacillus cereus WZZ006 was purified to homogeneity with an 89.5-fold purification, specific activity of 1.79 U/mg, and 26.7% recovery. The estimated molecular weight of BCE was 96 kDa which was analyzed by SDS-PAGE and MALDI-TOF-MS. Activity staining denotes that BCE has an unexplored new carboxyl esterase characteristic. BCE enzyme activity was maximum at pH 8.5 and also at 50 °C with pNP-caproate as a substrate. This indicates that the studied BCE as a thermoalkaliphilic esterase. The kinetic properties like Km, Vmax, kcat and kcat/Km value for BCE was found to be 0.98 mM, 0.03 mM/min, 69.47 min−1 and 70.89 mM−1 min−1, respectively. Synthesis of (S)-5-chloro-1-oxo-2,3-dihydro-2-hydroxy-1H-indole-2-carboxylic acid methyl ester ((S)-CODHCM) by BCE can be shortened to 3 h compared to 36 h with whole-cell catalysis. The e.e.s achieved was 93.83%, and conversion around 52.78% with E being 39.95. These features render BCE as a promising biocatalyst for the synthesis of a key chiral intermediate for indoxacarb.

Protein chains contain only l-amino acids, with the exception of the achiral glycine, making the chains homochiral. This homochirality is a prerequisite for proper protein folding and, hence, normal cellular function. The importance of d-amino acids as a component of the bacterial cell wall and their roles in neurotransmission in higher eukaryotes are well-established. However, the wider presence and the corresponding physiological roles of these specific amino acid stereoisomers have been appreciated only recently. Therefore, it is expected that enantiomeric fidelity has to be a key component of all of the steps in translation. Cells employ various molecular mechanisms for keeping d-amino acids away from the synthesis of nascent polypeptide chains. The major factors involved in this exclusion are aminoacyl-tRNA synthetases (aaRSs), elongation factor thermo-unstable (EF-Tu), the ribosome, and d-aminoacyl-tRNA deacylase (DTD). aaRS, EF-Tu, and the ribosome act as "chiral checkpoints" by preferentially binding to l-amino acids or l-aminoacyl-tRNAs, thereby excluding d-amino acids. Interestingly, DTD, which is conserved across all life forms, performs "chiral proofreading," as it removes d-amino acids erroneously added to tRNA. Here, we comprehensively review d-amino acids with respect to their occurrence and physiological roles, implications for chiral checkpoints required for translation fidelity, and potential use in synthetic biology.

Glycoside hydrolase 13 (GH13) family represents a large and diverse enzyme family. Cqm1, an amylomaltase of Culex mosquito, belongs to the GH13 family and subfamily 17 (GH13_17). The protein acts as the receptor for mosquito-larvicidal BinAB toxin that is used world-wide for control of the mosquito population. The protein was crystallized in the presence of a mixture of divalent metal ions. Cqm1 crystal structure was solved using the MRSAD method using Cd(II) anomalous at 1.9 A wavelength and the structure was refined against 1.8 A synchrotron data. One tightly bound Ca(II) ion in each of the monomer was observed and this site is suggested here to be unique to the GH13_17 family. Molecular dynamics simulations provide clues for the functional role of Ca(II) ion shown earlier to be essential for enzymatic activity. An optimized substrate (maltotriose) bound structure of the complex was constructed based on which ‘retaining-type’ mechanism can be predicted reliably. It reveals large conformational change in aromatic residues situated at active-site entrance. A Cd(II) ion was observed overlapping with the substrate-binding site. Kinetics data suggests non-competitive inhibition of Cqm1 by Cd(II). This is the first structure from the GH13_17 family and provides template for constructing reliable models for other members.

The enantioselective effects of imazethapyr (IM) enantiomers on wheat seedlings in a hydroponic medium were studied. R-IM at 0.05mg/L exerted a stronger inhibitory effect on shoot weight and root weight than 0.05mg/L S-IM, suggesting that R-IM more severely inhibited growth. Oxidative damage, based on the anthocyanin content, malondialdehyde (MDA) content, antioxidant enzyme activities and transcript levels of antioxidant enzyme genes, were studied together with the cellular ultrastructure of wheat leaves. The anthocyanin and MDA contents in the R-IM treatment group were significantly increased compared with those in the control group, but no significant changes were observed in the S-IM treatment group. The antioxidant enzyme activities of CAT and SOD were inhibited by 0.32- and 0.73-fold, respectively, in the 14day R-IM treatment group compared to those in the control. However, the transcript levels of antioxidant enzyme genes, including CuZnSOD, POD and CAT, were downregulated in the 14day R-IM exposure group, but those of DHAR were not. The number and size of starch granules increased and chloroplast swelling was observed in wheat leaf cells after R-IM exposure, which showed that photosynthetic functions were potentially disturbed. These results directly or indirectly imply that R-IM exposure causes more oxidative stress and exerts a stronger negative effect on wheat than S-IM. A metabolomics approach revealed that the tricarboxylic acid cycle was heavily suppressed by R-IM treatment. Some amino acids (proline, threonine, lysine, valine) were increased by only the R-IM treatment, indicating the activation of antioxidant pathways. The decrease in a series of fatty acids implied that the cell membrane composition changed in response to R-IM. These results provide a deeper understanding of the enantioselective effects of IM enantiomers on the molecular and metabolic responses in wheat seedlings.

In this work, molecularly imprinted electrospun chromogenic membrane(MIP-ECM) was synthesized using P-Hydroxybenzene propanoic acid as imitate template molecule and Poly (vinyl alcohol)(PVA) as substrate. Ninhydrin is used as chromogenic agent for visualizing detection of target molecule l-tyrosine(Tyr). The results showed that molecularly imprinted membrane exhibited selectivity and specific adsorption capacity for l-tyrosine. The influences of environmental factors, such as reaction temperature, the amount of ninhydrin and elution time, on chromogenic properties were examined by SEM and optical microscope. Gray value analysis is proceeded by software Image J. Due to the facilitate preparation and intuitive visualization, the proposed method was a promising candidate for selective recognition of bio-generated acid and amino molecules.

Pseudopaline and staphylopine are opine metallophores biosynthesized by Pseudomonas aeruginosa and Staphylococcus aureus, respectively. The final step in opine metallophore biosynthesis is the condensation of the product of a nicotianamine (NA) synthase reaction (i.e. l-HisNA for pseudopaline and d-HisNA for staphylopine) with an α-keto acid (α-ketoglutarate for pseudopaline and pyruvate for staphylopine), which is performed by an opine dehydrogenase. We hypothesized that the opine dehydrogenase reaction would be reversible only for the opine metallophore product with (R)-stereochemistry at carbon C2 of the α-keto acid (prochiral prior to catalysis). A kinetic analysis using stopped-flow spectrometry with (R)- or (S)-staphylopine and kinetic and structural analysis with (R)- and (S)-pseudopaline confirmed catalysis in the reverse direction for only (R)-staphylopine and (R)-pseudopaline, verifying the stereochemistry of these two opine metallophores. Structural analysis at 1.57–1.85 Å resolution captured the hydrolysis of (R)-pseudopaline and allowed identification of a binding pocket for the l-histidine moiety of pseudopaline formed through a repositioning of Phe-340 and Tyr-289 during the catalytic cycle. Transient-state kinetic analysis revealed an ordered release of NADP(+) followed by staphylopine, with staphylopine release being the rate-limiting step in catalysis. Knowledge of the stereochemistry for opine metallophores has implications for future studies involving kinetic analysis, as well as opine metallophore transport, metal coordination, and the generation of chiral amines for pharmaceutical development.

Agricultural chemicals are normally used as mixtures of several isomers, e.g. , enantiomers. In theory, in order to minimize the pesticides dose, it is desirable to use the most target-active isomer. Metolachlor is a typical multichiral herbicide belonging to amide herbicides. An asymmetric carbon atom and a chiral axis yield four stereoisomers. In this study, a novel laboratory method was developed to prepare the S -metolachlor and the four stereoisomers using high performance liquid chromatography. The separated isomers had a purity of >99%, with their absolute configurations assigned by electronic circular dichroism. The enantioseparation by ultra performance convergence chromatography tandem mass spectrometry was also performed for the rapid and sensitive detection of metolachlor stereoisomers. The enantioselective herbicidal activity toward the target weed ( Echinochloa crusgalli ) was systematically assessed for the first time by measuring the morphology of the weed after treatment with rac -, S -metolachlor and the four stereoisomers, respectively. Among the commercial pesticides, S -metolachlor was more effective in weed inhibition than rac -metolachlor, and to the four stereoisomers, the herbicidal activities were ranked as: SS > SR ≫ RS > RR , and the RR -isomer even had some stimulative effect to the weed growth at lower concentration (1 ppm). Thus, we concluded that in these cases, the chiral carbon feature played a major role in herbicidal activity rather than the chiral axis feature, and the higher bioactivity of the S -isomers was confirmed by more effective uptake and stronger interaction with target enzymes that were involved in the gibberellic acid biosynthesis. Although the SS -isomer shows the highest herbicidal activity, controlling the major chiral feature is still much easier and more economical than controlling two chiral features.

The mechanisms of the tandem sequential [4 + 2]/[3 + 2] and [3 + 2]/[4 + 2] cycloaddition sequences involving an ester, cyclooctatetraene (COTE), and cyclic and acyclic nitrones for the formation of a diverse range of isoxazolidine derivatives and other synthetic precursors are reported. A thorough exploration of the PES has characterized several regio-, stereo- and enantio-selective mechanistic channels involved in these reactions. A perturbation molecular orbital (PMO) analysis been employed to rationalize the results. It has also been found that the initial electrocyclic ring closure of the COTE is the rate-determining step in the tandem sequential [4 + 2]/[3 + 2] addition sequence. The thermolytic breakdown of the tandem adducts to subsequent monocyclic, bicyclic and tricyclic adducts occurs generally with very high activation barriers making it an inconvenient synthetic approach. The different reactivity of all the three double bonds present in the dipolarophile is reported. Finally, the mechanistic possibilities of [3 + 2]/[4 + 2] addition sequences involving the same reaction components in the case of cyclic and acyclic nitrones are explored extensively. The results suggest a novel and convenient routes for obtaining products of high selectivity with less energetic requirements. In some instances, new cycloadducts hitherto unreported are obtained.