Amino Acid Research Articles

Flue-cured tobacco intercropping with insectary floral plants improves rhizosphere soil microbial communities and chemical properties of flue-cured tobacco

BackgroundContinuous cropping of the same crop leads to land degradation. This is also called the continuous-cropping obstacle. Currently, intercropping tobacco with other crops can serve as an effective strategy to alleviate continuous cropping obstacles.ResultsIn this study, tobacco K326 and insectary floral plants were used as materials, and seven treatments of tobacco monoculture (CK), tobacco intercropped with Tagetes erecta, Vicia villosa, Fagopyrum esculentum, Lobularia maritima, Trifolium repens, and Argyranthemum frutescens respectively, were set up to study their effects on rhizosphere soil chemical properties and composition and structure of rhizosphere soil microbial community of tobacco. The 16 S rRNA gene and ITS amplicons were sequenced using Illumina high-throughput sequencing. tobacco/insectary floral plants intercropping can influence rhizosphere soil chemical properties, which also change rhizosphere microbial communities. The CK and treatment groups tobacco rhizosphere soil microorganisms had significantly different genera, such as tobacco intercropping with T. repens and A. frutescens significantly increased the number of Fusarium and intercropping T. erecta, V. villosa, L. maritima, T. repens, and A. frutescens significantly increased the number of Sphingomonas and unknown Gemmatimonadaceae. Additionally, intercropping T. erecta, V. villosa and L. maritima changed the rhizosphere fungal and bacteria community and composition of tobacco and the positive correlation between tobacco rhizosphere the genera of fungi and bacterial were greater than CK. The pathway of the carbohydrate metabolism, amino acid metabolism, and energy metabolism in rhizosphere bacteria were significantly decreased after continuous cropping. Fungal symbiotic trophic and saprophytic trophic were significantly increased after intercropping V. villosa, L. maritima and plant pathogen and animal pathogen were increased after intercropping T. repens and A. frutescens. Additionally, bacterial and fungal communities significantly correlated with soil chemical properties, respectively.ConclusionThis study reveals that intercropping tobacco with insectary floral plants, particularly T. erecta, V. villosa, L. maritima and A. frutescens significantly affects soil chemical properties and alters rhizosphere microbial communities, increasing the abundance of certain microbial genera. Additionally, intercropping enhances pathways related to carbohydrate, amino acid, and energy metabolism in rhizosphere bacteria. These findings suggest that intercropping could provide a promising strategy to overcome challenges associated with continuous tobacco cropping by regulating the rhizosphere environment.

Optimizing Probiotic Low‐Fat Yogurt: The Benefits of Incorporating Defatted Rice Bran for Enhanced Quality and Functionality

ABSTRACTThis study investigated the effects of incorporating defatted rice bran (DRB) at different concentrations (0.5%, 1%, 1.5%, and 2%) on the quality, microbiological, and sensory characteristics of probiotic low‐fat yogurt (LFY) during a 21‐day storage period at 4°C. DRB is rich in dietary fiber, essential amino acids, vitamins, and bioactive compounds, and its addition aimed to enhance the nutritional and functional properties of LFY. LFY samples were evaluated for proximate composition, hardness, viscosity, syneresis, color characteristics, rheology, and sensory evaluation. Also, microbiological analysis includes total bacterial counts (TBC), Lactobacillus delbrueckii ssp. bulgaricus, Streptococcus thermophilus, Bifidobacterium bifidum counts. Results indicated that DRB addition significantly (p < 0.05) increased probiotic counts, especially at 1% and 1.5% concentrations, with these samples maintaining higher bacterial stability over the storage period. DRB‐added LFY also showed improved nutritional profiles, with increased TS, protein, ash, fiber, and antioxidant activity. Additionally, hardness, viscosity, and rheology (G′ and G″) values significantly (p < 0.05) increased with the addition of DRB, while syneresis significantly (p < 0.05) decreased. However, higher DRB concentrations negatively affected the color, with lightness decreasing and the browning index increasing. This also impacted sensory characteristics, resulting in lower scores for color, flavor, and overall acceptability, particularly in LFY with 2% DRB. To conclude, moderate DRB addition (up to 1.5%) optimally balances the enhancement of probiotic and nutritional properties with acceptable sensory quality offering a viable strategy for producing functional low‐fat yogurt.

Hierarchical Structures of Amino Acid Derived Polyhydroxyurethanes: Promising Candidates as Drug Carriers and Cell Adhesive Scaffolds.

In this study, we examined the self-assembly of a series of biodegradable and biocompatible amino acid-based polyhydroxyurethanes (PHUs), investigating the structural influence of these polymers on their self-assembly and the resulting morphological features. The presence of hydrophilic and hydrophobic segments, along with carbonyl urethane, ester, and hydroxyl groups in the PHU backbone, facilitates intermolecular hydrogen bonding, enabling the formation of self-assemblies with hierarchical nanodimensional morphologies. We determined the critical aggregation concentration (CAC) and found that it largely depends on the PHU's structure. In-depth morphological studies demonstrated that the evolution of morphology proceeds in four steps: (1) the initial formation of micelles, which act as seeds at very low concentrations, (2) the elongation of these micelles into nanorod or nanopalette shapes below the CAC range, (3) the epitaxial growth of nanofibers at the CAC, and (4) the complete formation of fibrous mats above the CAC. Additionally, these hierarchical structures were utilized for the encapsulation and release of the drug doxorubicin (DOX). We observed that 75% of the encapsulated DOX was readily released in a mildly acidic environment, similar to the physiological conditions of cancer cells. Cellular uptake studies confirmed the effective uptake of the drug-loaded nanoassemblies into the cytoplasm of cells. Our studies also confirmed that these self-assembled structures can serve as effective cell adhesive scaffolds for tissue engineering applications.

Structure and functionality of Pleurotus geesteranus protein isolate as a function of pH.

Edible mushroom proteins hold great potential for food applications, but those extracted using the alkaline extraction-acid precipitation method typically exhibit poor solubility in neutral water, with the structural changes during acid precipitation remaining unclear. In this study, Pleurotus geesteranus protein isolate (PGPI) with high water solubility was prepared with alkaline extraction, followed by dialysis and freeze-drying, and the effects of pH on the structural and functional properties of PGPI were systematically investigated. PGPI was enriched in essential and aromatic amino acids, and the molecular weight of bands in the sodium dodecyl sulfate-polyacrylamide gel electrophoresis profile was mainly distributed below 45kDa. The zeta potential of PGPI changed from +16.84 to 17.58mV when the pH increased from 2 to 9, with a pI of 4.3. At pH 7, PGPI showed a size of 232.7nm. Away from pH 7, the particle size of PGPI increased. When the pH decreased from 7 to 2, PGPI exhibited a lower α-helix structure content and a higher β-sheet content and a gradual decrease in fluorescence intensity. In addition, as the pH approached 4, H0 and the content of SS group increased to a peak. These results indicated that lowering the pH induced the development of more ordered protein structure, which could be the primary reason for the poor water solubility of P. geesteranus protein obtained through alkaline extraction and acid precipitation. Additionally, these structural changes result in alterations to its functional properties, including water-holding capacity, oil-holding capacity, foaming capacity, foaming stability, emulsion activity index, and emulsion stability index.

Effects of nanoplastics on the growth, transcription, and metabolism of rice (Oryza sativa L.) and synergistic effects in the presence of iron plaque and humic acid

Nanoplastics (NPs) can adversely affect living organisms. However, the uptake of NPs by plants and the physiological and molecular mechanisms underlying NP-mediated plant growth remain unclear, particularly in the presence of iron minerals and humic acid (HA). In this study, we investigated NP accumulation in rice (Oryza sativa L.) and the physiological effects of exposure to polystyrene NPs (0, 20, and 100 mg L−1) in the presence of iron plaque (IP) and HA. NPs were absorbed on the root surface and entered cells, and confocal laser scanning microscopy confirmed NP uptake by the roots. NP treatments decreased root superoxide dismutase (SOD) activity (28.9–44.0%) and protein contents (31.2–38.6%). IP and HA (5 and 20 mg L−1) decreased the root protein content (20.44–58.3% and 44.2–45.2%, respectively) and increased the root lignin content (22.3–27.5% and 19.2–29.6%, respectively) under NP stress. IP inhibited the NP-induced decreasing trend of SOD activity (19.2–29.5%), while HA promoted this trend (48.7–50.3%). Transcriptomic and metabolomic analysis (Control, 100NPs, and IP-100NPs-20HA) showed that NPs inhibited arginine biosynthesis, and alanine, aspartate, and glutamate metabolism and activated phenylpropanoid biosynthesis related to lignin. The coexistence of IP and HA had positive effects on the amino acid metabolism and phenylpropanoid biosynthesis induced by NPs. Regulation of genes and metabolites involved in nitrogen metabolism and secondary metabolism significantly altered the levels of protein and lignin in rice roots. These findings provide a scientific basis for understanding the environmental risk of NPs under real environmental conditions.

Rapid production of recombinant rotaviruses by overexpression of NSP2 and NSP5 genes with modified nucleotide sequences.

Reverse genetics systems for rotaviruses (RV) facilitate the generation of genetically engineered RVs by transfection of 11 plasmids encoding 11 genomic viral RNA segments. In addition to viral genome expression, overexpression of NSP2 and NSP5 has been used to increase the rescue efficiency of recombinant RVs. Here, we showed that the overexpression of nucleotide sequence-modified NSP2 and NSP5 enabled the rapid and efficient production of recombinant RVs. Using improved reverse genetics, we established a reverse genetics system for human and bovine RV clinical isolates, as well as laboratory strains of bovine RV (NCDV and UK) and porcine RV (Gottfried). In addition, we rescued low-replicating recombinant RVs carrying a mutant NSP4 lacking the double-layered particle-binding domain, which was deficient in the efficient production of mature virions. These advancements in reverse genetics enabled the generation of molecular clones of RV clinical isolates and recombinant RVs harboring critical amino acid mutations, offering a versatile platform for investigating RV biology and pathogenesis.IMPORTANCERecombinant rotavirus (RV) synthesis via reverse genetics relies on both the viral propagation capacity and the efficiency of the experimental system. Since the establishment of our reverse genetics system, several enhancements have been implemented to augment the rescue efficiency. Nevertheless, challenges persist in generating RV clinical strains and recombinant viruses with low replication capacities. Notably, this improved reverse genetics system successfully facilitated the establishment of molecular clones of human and bovine RV clinical isolates. Fecal samples from patients with RV typically harbor quasi-species or, occasionally, multiple genotypes of RV. In the present study, we performed the genetic sequencing of clinical viral strains during the early propagation stages in cultured cells. Subsequently, infectious viruses were synthesized, allowing the characterization of circulating viruses in nature. This approach provides valuable insights into the genetic diversity and dynamics of RV populations and contributes to a more comprehensive understanding of viral pathogenesis and evolution.

Beyond soybean meal: investigating the effects of dietary protein alternatives on gut health, liver function and microbiota in traditional slow-growing chicken breeds

The use of soybean meal in animal feed is crucial for nutrition but has a significant environmental impact, especially related to deforestation and biodiversity loss. With the increasing demand for sustainable agricultural practices, it becomes essential to explore alternatives to soybean meal. This study evaluated the effect of a soy-free diet, replaced with fava beans and pea protein, on an Italian slow-growing chicken breed: the Bianca di Saluzzo. The results indicate that the experimental diet did not compromise the growth, final weight or intestinal health of the animals, demonstrating a good tolerance and adaptability of the breed to these alternative ingredients. Furthermore, the analysis of the intestinal microbiota showed a positive impact, with an increase in organic acids such as succinic and citric, which could improve intestinal health and metabolic efficiency. Some changes in liver enzyme levels were observed, such as the increase in glutamate-oxaloacetate transaminase, an enzyme involved in amino acid metabolism. This increase could indicate a higher efficiency in protein metabolism, suggesting that the diet based on alternative ingredients could support an improvement in liver metabolism. Although this aspect deserves further investigation, it could represent a positive effect of the diet on liver function and overall health of the animals. Further studies are needed to fully understand the long-term implications of these dietary modifications, particularly in relation to the gut microbiota and metabolism.

Functional analysis of a S-adenosylmethionine-insensitive methylenetetrahydrofolate reductase identified in methionine-accumulating yeast mutants.

Essential amino acids (EAAs) are important for the maintenance of brain functions. Therefore, the yeast Saccharomyces cerevisiae that accumulate EAAs would help elderly people ingest appropriate levels of EAAs, which in turn could slow neurodegeneration, extend the healthy lifespan and improve quality of life. Here, we isolated two mutant strains, ETH-80 and ETH-129, that accumulate the EAA methionine. Both strains were derived from a diploid laboratory yeast by conventional mutagenesis and carry a novel mutation in the MET13 gene, which encodes the Ser443Phe variant of methylenetetrahydrofolate reductase. Enzymatic analysis revealed that the Ser443Phe substitution abolished the sensitivity to S-adenosyl methionine (SAM)-mediated inhibition even in the presence of 2mM SAM, while increasing the activity for NADPH-dependent reduction. Furthermore, yeast cells expressing the Ser443Phe variant showed a fourfold increase in intracellular methionine content compared to the wild-type Met13. These findings will be useful for the future development of methionine-accumulating yeast strains.

L858R/L718Q and L858R/L792H Mutations of EGFR Inducing Resistance Against Osimertinib by Forming Additional Hydrogen Bonds.

Acquired resistance to first-line treatments in various cancers both promotes cancer recurrence as well as limits effective treatment. This is true for epidermal growth factor receptor (EGFR) mutations, for which secondary EGFR mutations are one of the principal mechanisms conferring resistance to the covalent inhibitor osimertinib. Thus, it is very important to develop a deeper understanding of the secondary mutational resistance mechanisms associated with EGFR mutations arising in tumors treated with osimertinib to expedite the development of innovative therapeutic drugs to overcome acquired resistance. This work uses all-atom molecular dynamics (MD) simulations to investigate the conformational variation of two reported EGFR mutants (L858R/L718Q and L858R/L792H) that resist osimertinib. The wild-type EGFR kinase domain and the L858R mutant are used as the reference. Our MD simulation results revealed that both the L718Q and L792H secondary mutations induce additional hydrogen bonds between the residues in the active pocket and the residues with the water molecules. These additional hydrogen bonds reduce the exposure area of C797, the covalent binding target of osimertinib. The additional hydrogen bonds also influence the binding affinity of the EGFR kinase domain by altering the secondary structure and flexibility of the amino acid residues in the domain. Our work highlights how the two reported mutations may alter both residue-residue and residue-solvent hydrogen bonds, affecting protein binding properties, which could be helpful for future drug discovery.

Structure and function of the proton channel OTOP1

OTOP1 belongs to the otopetrin family of membrane proteins that form proton channels in cells of diverse types. In mammals, OTOP1 is involved in sour transduction in taste cells and contributes to otoconia formation in the inner ear. From the structural point of view, otopetrins, including OTOP1, represent a quasi-tetramer consisting of four α-barrels. The exact transport pathways mediating proton flux through the OTOP1 channel and gating units modulating its activity are still a matter of debate. This review discusses current data on structural and functional features of OTOP1. Suggested proton transport pathways, regulatory mechanisms, and key amino acid residues determining functionality of the otopetrins are considered. The existing kinetic models of OTOP1 are discussed as well. Based on revealed functional properties, OTOP1 is suggested to operate as a logical XOR element that allows for proton flux only if transmembrane pH gradient exists.

Identification of Olea europaea CBF/DREB1 Family Genes in Abnormal Temperature Stress Response

Olea europaea, native to the Mediterranean region, has been widely cultivated for the nutritional content of its fruits and leaves. To adapt to climates beyond the Mediterranean, research on the climatic adaptation of O. europaea is urgently needed. Therefore, in this study, the CBF/DREB1 family genes in olives, which are related to cold tolerance, were genome-wide characterized. In total, four OeCBFs were screened; their open reading frame (ORF) were 552~684 bp in length, the encoded proteins were 20,786.60~25,235.16 Da, with 183~227 amino acids, and the theoretical isoelectric point (pI) were 4.91~9.13. The transcription of four OeCBFs was significantly different in olive tissues, especially OeCBF2 and OeCBF4, which were substantially more highly-expressed in aboveground tissues (terminal buds, young leaves, old leaves, and stems) than in underground tissues (thick and fine roots). Exposed to cold stress, OeCBF2 and OeCBF4 were up-regulated more obviously in olive leaves and stems, while OeCBF3 was induced to higher levels in the roots, implying the adaptation potential of the OeCBFs to low temperature stress. A comparison of the expression of the four OeCBFs in two varieties, Picholine (more tolerant to the other one) and Arbequina, found that the expression of OeCBFs was higher in most tissues of Picholine than that of Arbequina. When exposed to cold stress, the expression of the OeCBFs was also higher in the leaves and roots of Picholine than in Arbequina, further confirming the correlation between OeCBFs and cold tolerance of olive trees. This study not only deepens our understanding of the CBF gene family in olives, but also provides significant genetic information for olive breeding and improvement.

Zinc oxide nanoparticles cooperate with the phyllosphere to promote grain yield and nutritional quality of rice under heatwave stress

To address rising global food demand, the development of sustainable technologies to increase productivity is urgently needed. This study revealed that foliar application of zinc oxide nanoparticles (ZnO NPs; 30 to 80 nm, 0.67 mg/d per plant, 6 d) to rice leaves under heatwave (HW) stress increased the grain yield and nutritional quality. Compared with the HW control, the HWs+ZnO group presented increases in the grain yield, grain protein content, and amino acid content of 22.1%, 11.8%, and 77.5%, respectively. Nanoscale ZnO aggregated on the leaf surface and interacted with leaf surface molecules. Compared with that at ambient temperature, HW treatment increased the dissolution of ZnO NPs on the leaf surface by 25.9% and facilitated their translocation to mesophyll cells. The Zn in the leaves existed as both ionic Zn and particulate ZnO. Compared with the HW control, foliar application of ZnO NPs under HW conditions increased leaf nutrient levels (Zn, Mn, Cu, Fe, and Mg) by 15.8 to 416.9%, the chlorophyll content by 22.2 to 24.8%, Rubisco enzyme activity by 21.2%, and antioxidant activity by 26.7 to 31.2%. Transcriptomic analyses revealed that ZnO NPs reversed HW-induced transcriptomic dysregulation, thereby enhancing leaf photosynthesis by 74.4%. Additionally, ZnO NPs increased the diversity, stability, and enrichment of beneficial microbial taxa and protected the phyllosphere microbial community from HW damage. This work elucidates how NPs interact with the phyllosphere, highlighting the potential of NPs to promote sustainable agriculture, especially under extreme climate events (e.g., HWs).

DNA-Encoded Noncanonical Substrate Library for Protease Profiling.

Profiling the substrate sequence preferences of proteases is important for understanding both biological functions as well as for designing protease inhibitors. Several methods are available for profiling the sequence specificity of proteases. However, there is currently no rapid and high-throughput method to profile specificity of proteases for noncanonical substrates. In this study, we described a strategy to use a DNA-encoded noncanonical substrate library to identify the protease substrates composed of both canonical and noncanonical amino acids. This approach uses a DNA-encoded peptide library and introduces a biotin molecule at the N-terminus to immobilize the library on a solid support. Upon protease hydrolysis, the released DNA tag of the substrate peptides can be sequenced to identify the substrate structures. Using this approach, we profiled trypsin and fibroblast activation protein α and discovered noncanonical substrates that were more efficiently cleaved than the commonly used substrates. The identified substrates of FAP were further used to design corresponding covalent inhibitors containing non-canonical sequences with high potency for the target protease. Overall, our approach can aid in the development of new protease substrates and inhibitors.

TCellR2Vec: efficient feature selection for TCR sequences for cancer classification

Cancer remains one of the leading causes of death globally. New immunotherapies that harness the patient’s immune system to fight cancer show promise, but their development requires analyzing the diversity of immune cells called T-cells. T-cells have receptors that recognize and bind to cancer cells. Sequencing these T-cell receptors allows to provide insights into their immune response, but extracting useful information is challenging. In this study, we propose a new computational method, TCellR2Vec, to select key features from T-cell receptor sequences for classifying different cancer types. We extracted features like amino acid composition, charge, and diversity measures and combined them with other sequence embedding techniques. For our experiments, we used a dataset of over 50,000 T-cell receptor sequences from five cancer types, which showed that TCellR2Vec improved classification accuracy and efficiency over baseline methods. These results demonstrate TCellR2Vec’s ability to capture informative aspects of complex T-cell receptor sequences. By improving computational analysis of the immune response, TCellR2Vec could aid the development of personalized immunotherapies tailored to each patient’s T-cells. This has important implications for creating more effective cancer treatments based on the individual’s immune system.

Zinc bioinspired catalytic system for the valorisation of CO2 into cyclic carbonates

Cyclic organic carbonates are defined as key compounds for a sustainable chemical economy. Their synthesis from CO2 under mild conditions is a useful way to valorise this greenhouse gas as carbon source. Even if a wide range of catalysts were described to promote the carbon dioxide cycloaddition into epoxides, only few ones concern enzymatic systems. The zinc/L‐histidine active site of carbonic anhydrase inspired the present work, pointing out that the imidazole moiety of the amino acid ligand has a crucial role. An extensive study was undertaken to establish the structure‐activity relationship of imidazole derivatives, zinc salts and their respective catalytic activity in the CO2 cycloaddition reaction. The effect of aromatic, alkyl or iodine substituents and their position in N‐heterocycles were highlighted. A synergic effect was noted when combining imidazole compounds with zinc salts. The optimisation of reaction conditions emphasised the in‐situ ZnI2/1‐methylimidazole catalytic system is selective towards cyclic styrene carbonates and efficient under solvent‐free mild conditions (50°C, atmospheric CO2 pressure). Once reusing tests confirmed the catalytic system robustness, the reaction scope was enlarged to several epoxides resulting in 84‐99% yields of their corresponding cyclic carbonates.

Molecular Evolution and Adaptation Strategies in Marine Ciliates: An Inspiration for Cold-Adapted Enzyme Engineering and Drug Binding Analysis

In the present review, we summarize genome mining of genomic data obtained from the psychrophilic Antarctic marine ciliate Euplotes focardii and its evolutionary-close mesophilic cosmopolitan counterpart E. crassus. This analysis highlights adaptation strategies that are unique to the Antarctic ciliate, including antioxidant gene duplication and distinctive substitutions that may play roles in increased drug binding affinity and enzyme reaction rate in cold environments. Enzymes from psychrophiles are usually characterized by high activities and reaction rates at low temperatures compared with their counterparts from mesophiles and thermophiles. As a rule, catalyst cold activity derives from an increased structural flexibility that may lead to protein denaturation in response to temperature fluctuation. Molecular thermolability has been a major drawback of using macromolecules from psychrophiles in industrial applications. Here, we report a case study in which the role of peculiar amino acid substitution in cold adaptation is demonstrated by site-directed mutagenesis. Combined with a rational design approach, these substitutions can be used for site-directed mutagenesis to obtain cold-active catalysts that are structurally stable. Furthermore, molecular docking analysis of β-tubulin isotypes extrapolated from E. focardii and E. crassus genomes allowed us to obtain additional insight on the taxol binding site and drug affinity. E. focardii genome mining and the comparison with the mesophilic sibling counterpart can be used as an inspiration for molecular engineering for medical and industrial applications.

Chemical profiling, in-silico investigation and in vivo toxicity assessment of lacatomtom (a psychoactive mixture) on selected indices in albino wistar rats

aims: To evaluate the safety and identify the psychoactive constituent of a new psychoactive cocktail that is abused among African youths. background: The use of lacatomtom (LTT), a psychoactive mixture of tomtom (TT) candies with lacasera (LC) beverage, has recently increased among young Nigerians and Africans. There isn't much scientific study on the constituent and effects of this psychoactive substance. objective: To chemically-profile the mixture using GCMS analysis, and evaluate the effect of the constituent of neural targets. To evaluate the toxicological effects in albino rats. To identify neur method: Herein, LTT was chemically-profiled using GCMS analysis, and the toxicological effects were examined in albino rats. In vivo experiment consists of five groups of six rats each (group 2 - LTT ad libitum; groups 1, 3, & 4 - TTT, TT, LC (1 mL) mg/mL kg/body weight once/day respectively, group 5 - distilled water ad libitum). Identified constituents were examined against human monoamine oxidase (hMOA) and human catechol O-methyltransferase (hCOMT) using in silico methods. result: Forty-seven chemical compounds were identified. Ad libitum intake of LTT elevated plasma alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, creatinine, total cholesterol, and LDL-cholesterol levels. The docked poses, binding scores, and interactions with amino acids informed the selection of (4-Methoxymethoxy-hex-5-ynylidene)-cyclohexane (MM) (-9.4 kcal/mol) and 3-(hydroxyphenylmethyl)-3,4-dimethyl-1-phenylpentan-2-one (HP) for hCOMT (-9.4 kcal/mol), while propionylcodeine (-10.1 kcal/mol) and HP (-8.9 kcal/mol) for hMOA. Top-docked compounds (TDC) demonstrated the potential to permeate the blood-brain barrier. TDC was predicted to be a positive substrate of the P-glycoprotein and presents inhibitory potential for cytochrome P450 descriptors. HP was mutagenic and could induce human hepatotoxicity and drug-induced liver injury, while propionylcodeine had human hepatotoxic prediction. conclusion: The present study, for the first time, confirmed the potential toxicity of lacatomtom to the liver, kidney, heart, and central nervous system supported by the identified top-docked compounds regarded as potential psychoactive constituents of hMOA and hCOMT. other: Nono

Nitrogen deposition and heathland management cause multi‐element stoichiometric mismatches, reducing insect fitness

Abstract Increased nitrogen and acidifying deposition negatively affect biogeochemical cycles and nutrient stoichiometry, particularly in oligotrophic habitats. In heathlands, management aims to mitigate effects of deposition by topsoil removal (sod‐cutting) or by liming to increase soil pH. However, these management efforts may further exacerbate macro‐ and micronutrient stoichiometric mismatch, with cascading effects on heathland insects. Here, we investigate the physiological mechanisms by which deposition and management affect insects, focusing on nutrient uptake and excretion. We combined a phosphorus (P) addition and liming experiment in previously sod‐cut heathland with laboratory‐rearing experiments using the field cricket (Gryllus campestris L., 1) as a model species. We hypothesized that P addition would benefit cricket performance by reducing food N:P ratio. Conversely, we hypothesized that liming would reduce performance by decreasing food Mn:Mg and/or Fe:Mg ratios. Elemental uptake efficiency, consumption and reproduction, as well as cricket field density, were significantly higher under increased P availability, confirming insect P limitation in the field. Liming decreased food Mn:Mg ratio and reduced reproductive success, eliminating any beneficial effects of P addition. Moreover, cricket stoichiometry better explained growth rates than the amount of food consumed. The best model highlighted cricket N and Mn content, with increasing N content negatively influencing growth, while the opposite was true for Mn. We found no support for liming‐induced Fe limitation. We propose that both P and Mn limit crickets independently. Specifically, P limitation affects growth via reduced ribosomal content, and Mn limitation impacts growth by limiting arginase activity and hence restricting the recycling of amino acids. Our study shows that restoration management can fail to restore habitat quality for insects and may even amplify mismatches in macro and micronutrient stoichiometry. To address this, the addition of not only P but also Mn may be required as follow‐up management, but reducing N‐emission remains imperative as this removes the need for disruptive management interventions. Read the free Plain Language Summary for this article on the Journal blog.

Molecular dynamics simulations unveil the aggregation patterns and salting out of polyarginines at zwitterionic POPC bilayers in solutions of various ionic strengths

This study employs molecular dynamics (MD) simulations to investigate the adsorption and aggregation behavior of simple polyarginine cell-penetrating peptides (CPPs), specifically modeled as R9 peptides, at zwitterionic phosphocholine POPC membranes under varying ionic strengths of two peptide concentrations and two concentrations of NaCl and CaCl2. The results reveal an intriguing phenomenon of R9 aggregation at the membrane, which is dependent on the ionic strength, indicating a salting-out effect. As the peptide concentration and ionic strength increase, peptide aggregation also increases, with aggregate lifetimes and sizes showing a corresponding rise, accompanied by the total decrease of adsorbed peptides at the membrane surface. Notably, in high ionic strength environments, large R9 aggregates, such as octamers, are also observed occasionally. The salting-out, typically uncommon for short positively charged peptides, is attributed to the unique properties of arginine amino acid, specifically by its side chain containing amphiphilic guanidinium (Gdm+) ion which makes both intermolecular hydrophobic like-charge Gdm+ – Gdm+ and salt-bridge Gdm+ – C-terminus interactions, where the former are increased with the ionic strength, and the latter decreased due to electrostatic screening. The aggregation behavior of R9 peptides at membranes can also be linked to their CPP translocation properties, suggesting that aggregation may aid in translocation across cellular membranes.

High-throughput screening and identification of lignin peroxidase based on spore surface display of Bacillus subtilis.

Lignin peroxidase is closely related to agriculture and food as it improves the quality of feedstuffs, facilitates the degradation of lignin in agricultural wastes, and degrades azo dyes that have similar complex structures to lignin. However, the current status of homologous or heterologous expression of lignin peroxidase is unsatisfactory and needs to be modified with the help of immobilization and directed evolution to maximize its potential. Directed evolution technology is an effective strategy for designing and improving enzyme characteristics, and Bacillus subtilis spore surface display technology is an efficient method for preparing immobilized enzymes. A colorimetric dye decolorization assay using Congo red as a substrate was developed and optimized for high-throughput screening of spore surface display in a 96-well plate. After two rounds of screening, a superior mutant strain was selected from 2700 mutants. Its highest catalytic activity was 196.36%. The amino acid substitution sites were identified as N120D and I242T. The mechanism for the enhanced catalytic activity was explained using protein modeling and functional analysis software. This study provides insights into the rational design of lignin peroxidase and its application in food and agriculture. © 2024 Society of Chemical Industry.