Search

Background: Previous studies have identified structurally diverse N-acyl amino acid methyl esters (NAMEs) in culture extracts of Roseovarius tolerans EL-164 (Roseobacteraceae). NAMEs are structural analogues of the common signaling compounds N-acyl homoserine lactones (AHLs), but do not participate in AHL-mediated signaling. NAMEs show minor antialgal and antimicrobial activity, but whether this activity serves as the primary ecological role remains unclear., Results: To enable dose-dependent bioactivity-testing, we have established a chromatographic method for quantification of NAMEs in bacterial culture extracts. The concentrations determined for the two major NAMEs produced by EL-164, C16:1-NAME and C17:1-NAME, ranged between 0.685 and 5.731 mg L - 1 (2.0-16.9 µM) and 5.3-86.4 µg L - 1 (15.0-244.3 nM), respectively. Co-quantification of the C14:1-AHL showed concentrations ranging between 17.5 and 58.7 mg L - 1 (56.6-189.7 µM). We observed distinct production patterns for NAMEs and AHLs, with a continuous NAME production during the entire incubation period. We conducted a spike-in experiment, using the determined metabolite concentrations. By comparing the transcriptomes of pre- and post-metabolite-spikes, we identified three clusters of differentially expressed genes with distinct temporal expression patterns. Expression levels of stress response genes differed between NAME- and AHL-spiked EL-164 cultures in the stationary phase., Conclusions: Our findings support previous studies suggesting an ecological role for C16:1-NAME as antibiotic, by proving that NAME concentrations in batch cultures were higher than the minimal inhibitory concentrations against Maribacter sp. 62 - 1 (Flavobacteriia) and Skeletonema costatum CCMP 1332 (Coscinodiscophyceae) reported in the literature. Our study further exemplified the broad application range of dose-dependent testing and highlighted the different biological activities of NAMEs and AHLs., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Lysine residues on protein surfaces are abundant and often found in enzyme active sites, making them critical targets for studying undruggable proteins. However, the varied microenvironment surrounding lysine residues results in a wide range of p K a values, complicating site-specific covalent binding. In this study, we address the challenges posed by the diverse reactivity of amino side chains by modulating the amide reaction activity of heteroaromatic activated esters. By fine-tuning the type, position, and number of heteroatoms, we successfully rationalized the regulation of their amide reaction activity, leading to the design of probes for selective lysine labeling within the proteome for profiling purposes. Systematic optimization of these esters' reactivity and selectivity has yielded a series of effective probes suitable for both in vitro and cellular applications. These findings significantly enhance our understanding of protein functions and mechanisms, facilitated by the precise identification and analysis of protein labeling and profiling.

Hydroponics combined with fugacity model was employed to investigate the kinetics of uptake, accumulation, and metabolism of organophosphate esters (OPEs) by japonica rice. The time-dependent process for uptake and accumulation of 5 OPEs and their diester-metabolites in both rice root and shoot fitted well with the pseudo-first-order kinetic model. The peak OPE accumulations in rice root and shoot were significantly positively or negatively correlated with their octanol-water partition coefficient (logK ow ) respectively, but not for their apparent accumulation rates. Root concentration factors (RCFs) and root-to-shoot translocation factors (TFs) of OPEs were found to be positively and negatively correlated with their logK ow , respectively. Triphenyl phosphate with benzene ring substituents showed the highest RCF, but the lowest TF, because of its high potential for root adsorption due to the π electron-rich structures. Sterilized root exudates can hinder the root adsorption and absorption of OPEs from solution probably through competitive adsorption of OPEs with root surface. The first-hand transport and metabolism rates were also obtained by generating these rates to fit the dynamic fugacity model with the measurement values. The simulation indicated that the kinetics of OPE accumulation in rice plants may be controlled by multiple processes and physicochemical properties besides K ow ., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Geranyl esters (GEs) are valuable monoterpene esters derived from the esterification of geraniol and various carboxylic acids with a range of unique aromas and properties, making them valuable in perfumery, pharmaceutical, and cosmetic applications. Lipase-mediated esterification is considered to be a sustainable process but is challenged by the lack of a compatible catalytic method in conjunction with a customized microbial biosynthesis of geraniol. In this study, we developed an integrated process to convert glycerol and various carboxylates into GEs. The process includes microbial biosynthesis of geraniol using metabolically engineered Escherichia coli and enzymatic conversion of geraniol into GEs in a fermentation medium-organic biphasic system using an immobilized lipase. The enzymatic step for esterifying the target carboxylates with geraniol achieved >90% conversion under the optimized condition. Coupled with the geraniol from microbial fermentation, 0.59 g/L geranyl butyrate and 1.04 g/L geranyl hexanoate were produced subsequently, demonstrating the feasibility of converting renewable source into monoterpene esters through this integrated process, which bypassed feeding extra geraniol in the conventional lipase-mediated GE synthesis.

Organophosphate triesters (tri-OPEs) and diesters (di-OPEs) may threaten human health through dietary intake, whereas little information is available about their fate in mammals. Herein, mice exposure experiments were carried out through gavage with six tri-OPEs and six di-OPEs, respectively. The residual levels of di-OPEs in mice were generally higher than those of tri-OPEs. The residual di-OPEs mainly distributed in the liver and blood while the most tri-OPEs remained in stomach, indicating easier transfer and lower metabolism levels of di-OPEs. The accumulation of tri- and di-OPEs with large octanol-water partition coefficients and long carbon chain were observed in tissues and feces, implying that the elimination of these OPEs through fecal excretion is an important elimination pathway. A total of 86 OPE metabolites were found in murine urine and feces, 57 of which were identified for the first time. For tri-OPEs, carboxylated OPEs had higher peak intensities and fewer interference factors among the metabolites, which could serve as ideal biomarkers. The predicted oral median lethal doses of OPEs and corresponding metabolites showed an increased toxicity of some hydroxylated OPEs and di-OPEs, needing further attention. These results provided new insights and evidence on the fates and biomarkers of OPEs exposure for mammals., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The effect of fermentation strains (Lactiplantibacillus plantarum CQ01107 and Staphylococcus simulans CD207) on the binding properties of porcine myofibrillar proteins (MPs) to esters was investigated from two perspectives: metabolism degree and structural alterations. Results demonstrated that S. simulans could reduce the particle size and α-helix content of MPs, while simultaneously increasing the absolute zeta potential and active sulfhydryl content. This process decreased protein aggregation and facilitated the unfolding of MPs, thereby enhancing their binding to esters. Conversely, L. plantarum showed limited promotion, which might be related to its robust acid production and protein hydrolysis capacities. In addition, ethyl octanoate, with a longer carbon chain length, was found to have the highest binding capacity to MPs (28.38 %-41.59 %). Molecular docking results further revealed that the binding of the four esters to MPs was spontaneous, with ethyl octanoate exhibiting the lowest binding energy to MPs (-5.635 kcal/mol). The primary forces involved in the binding of the four selected esters to MPs were hydrophobic interactions, hydrogen bonding, and van der Waals forces. These findings can provide new insights into the mechanisms by which fermentation strains influence flavor formation in fermented foods., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Fungi of the genus Ceratocystis are aggressive tree pathogens that cause serious diseases in several crops around the world. Ceratocystis wilt disease caused by C. cacaofunesta has been shown to be responsible for severe reductions in cacao production. In this study, headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry (GC-MS) was used in combination with chemometric analysis for monitoring volatile organic compounds (VOCs) released from C. cacaofunesta. Low-molecular-weight esters, alcohols, ketones, and sulphur compounds were identified in the liquid broth. Monitoring the volatile profile over five days of fungal growth revealed that the concentrations of alcohol and esters were inversely proportional. Acetate esters were responsible for the intense fruity aroma of the C. cacaofunesta culture produced within the first hours after fungal inoculation, which decreased over time, and are likely associated with the attraction of insect vectors to maintain the life cycle of the pathogen. PCA revealed that 3-methylbutyl acetate was the metabolite with the highest factor loading for the separation of the VOC samples after 4 h of fungal growth, whereas ethanol and 3-methylbutan-1-ol had the highest factor loadings after 96 and 120 h. 3-Methylbutan-1-ol is a phytotoxic compound that is likely associated with host cell death since C. cacaofunesta is a necrotrophic fungus. Fungal VOCs play important roles in natural habitats, regulating developmental processes and intra- and interkingdom interactions. This is the first report on the volatiles released by C. cacaofunesta., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Serine hydrolases have become increasingly important for recycling PET plastics. However, their properties are inherently constrained by their 3D structure, which in turn limits the conditions for their application. Considering peptides as catalysts for industrial depolymerization processes can help us to escape some of these limitations. In this article, a 25 amino acid thermostable peptide, HSH-25, was designed to depolymerize PET. The peptide incorporates a His-Ser-His motif, inspired by the catalytic triad found in the serine hydrolase family, into a β-hairpin fold. Stability of the fold was investigated by molecular dynamics simulations. Esterolytic activity of the peptide toward model substrates was detected within a pH range from pH 7 to pH 9.5. Degradation of polymeric PET substrates was confirmed by atomic force microscopy imaging on spin-coated PET thin films.

Glucose esters, heralded for their emulsifying and solubilizing properties, have found increasing application in food, pharmaceutical, and cosmetic sectors. However, the environmental impact of chemical synthesis and the problem of enzyme inactivation in enzymatic synthesis have hindered their application. To this end, deep eutectic solvents (DESs) with stabilized enzyme activity and better solubility properties are considered a promising solution. In the study, 12 kinds of hydrophilic DESs and 22 kinds of hydrophobic DESs were screened, and we examined their efficiency in the synthesis of glucose esters. The results indicated that d,l-menthol-based DESs have excellent performance, with the highest yield of 92.85%. The molecular dynamics simulations suggested that it can be attributed to DESs changing the size of substrate binding pocket, which in turn affects the substrate anchoring ability and the catalytic efficiency of lipase. To further evaluate the effects of the solvent on the substrate anchoring ability of enzymes, a novel strategy, namely, substrate anchoring index, which is based on the stability of the tetrahedral intermediate, was proposed and used for the designing of more efficient DESs. Fortunately, novel DESs based on cyclohexanone (menthol analogues) were successfully developed with a yield of 98.85%.

Precisely controlling enzyme conformation to enhance catalytic performance is a highly sought-after yet challenging goal in the immobilization of biocatalysts. Excessively strong enzyme-carrier interactions can restrict enzyme dynamics and reduce catalytic efficiency, while excessively weak interactions may lead to enzyme leakage, thereby reducing reusability. In this study, we developed a novel strategy to finely regulate the interaction between the carrier and the enzyme through the adjustment of the ratio of amino and octadecyl functional groups. The expressed activity of the novel immobilized lipase, CRL@AOMR, was 1.32- and 2.34-fold higher than that of the monofunctional macroporous resin. Moreover, the synthesis of various phytosterol esters in solvent-free systems was conducted as a model reaction to investigate the utilization of CRL@AOMR in different reactions. Under optimized conditions, an impressive yield of 96.1% for phytosterol oleate was achieved and a yield of 76.2% was maintained even after six cycles of utilization (288 h). This study demonstrates the potential feasibility of developing immobilization strategies via dual modification of amino and alkyl groups, which is a potential general strategy for other enzymes with surface lysine.

Carboxylesterases serve as potent biocatalysts in the enantioselective synthesis of chiral carboxylic acids and esters. However, naturally occurring carboxylesterases exhibit limited enantioselectivity, particularly toward ethyl 3-cyclohexene-1-carboxylate (CHCE, S1), due to its nearly symmetric structure. While machine learning effectively expedites directed evolution, the lack of models for predicting the enantioselectivity for carboxylesterases has hindered progress, primarily due to challenges in obtaining high-quality training datasets. In this study, we devise a high-throughput method by coupling alcohol dehydrogenase to determine the apparent enantioselectivity of the carboxylesterase AcEst1 from Acinetobacter sp. JNU9335, generating a high-quality dataset. Leveraging seven features derived from biochemical considerations, we quantitively describe the steric, hydrophobic, hydrophilic, electrostatic, hydrogen bonding, and π-π interaction effects of residues within AcEst1. A robust gradient boosting regression tree model is trained to facilitate stereodivergent evolution, resulting in the enhanced enantioselectivity of AcEst1 toward S1. Through this approach, we successfully obtain two stereocomplementary variants, DR3 and DS6, demonstrating significantly increased and reversed enantioselectivity. Notably, DR3 and DS6 exhibit utility in the enantioselective hydrolysis of various symmetric esters. Comprehensive kinetic parameter analysis, molecular dynamics simulations, and QM/MM calculations offer insights into the kinetic and thermodynamic features underlying the manipulated enantioselectivity of DR3 and DS6., (© 2024. The Author(s).)

As a traceless, bioreversible modification, the esterification of carboxyl groups in peptides and proteins has the potential to increase their clinical utility. An impediment is the lack of strategies to quantify esterase-catalyzed hydrolysis rates for esters in esterified biologics. We have developed a continuous Förster resonance energy transfer (FRET) assay for esterase activity based on a peptidic substrate and a protease, Glu-C, that cleaves a glutamyl peptide bond only if the glutamyl side chain is a free acid. Using pig liver esterase (PLE) and human carboxylesterases, we validated the assay with substrates containing simple esters ( e.g. , ethyl) and esters designed to be released by self-immolation upon quinone methide elimination. We found that simple esters were not cleaved by esterases, likely for steric reasons. To account for the relatively low rate of quinone methide elimination, we extended the mathematics of the traditional Michaelis-Menten model to conclude with a first-order intermediate decay step. By exploring two regimes of our substrate → intermediate → product (SIP) model, we evaluated the rate constants for the PLE-catalyzed cleavage of an ester on a glutamyl side chain ( k cat / K M = 1.63 × 10 3 M -1 s -1 ) and subsequent spontaneous quinone methide elimination to regenerate the unmodified peptide ( k I = 0.00325 s -1 ; t 1/2 = 3.55 min). The detection of esterase activity was also feasible in the human intestinal S9 fraction. Our assay and SIP model increase the understanding of the release kinetics of esterified biologics and facilitate the rational design of efficacious peptide prodrugs.

Plant uptake of organic contaminants generally occurs through either root, gas-phase foliar, or particle-phase foliar uptake. Understanding these pathways is essential for food-system practitioners to reduce human exposures, and to clean contaminated-sites with phytoremediation. Herein, we conducted a field-based experiment using an improved specific exposure chamber to elucidate the uptake pathways of organophosphate esters, phthalates, and polycyclic aromatic compounds, and quantitatively assessed their contributions to organic contaminant accumulations in field-grown rice. For most target compounds, all three uptake pathways (root, foliar gas, and foliar particle uptakes) contributed substantially to the overall contaminant burden in rice. Compounds with lower octanol-water partition coefficients (K ow ) were more readily translocated from roots to leaves, and compounds with higher octanol-air partition coefficients (K oa ) tended to enter rice leaves mostly through particle deposition. Most compounds were mostly stored in the inner leaves (55.3-98.2 %), whereas the relatively volatile compounds were more readily absorbed by the waxy layer and then transferred to the inner leaves. Air particle desorption was a key process regulating foliar uptake of low-volatility compounds. The results can help us to better understand and predict the environmental fate of those contaminants, and develop more effective management strategies for reducing their human exposure through food ingestion., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

α-Amino esters are precursors to noncanonical amino acids used in developing small-molecule therapeutics, biologics, and tools in chemical biology. α-C-H amination of abundant and inexpensive carboxylic acid esters through nitrene transfer presents a direct approach to α-amino esters. Methods for nitrene-mediated amination of the protic α-C-H bonds in carboxylic acid esters, however, are underdeveloped. This gap arises because hydrogen atom abstraction (HAA) of protic C-H bonds by electrophilic metal-nitrenoids is slow: metal-nitrenoids preferentially react with polarity-matched, hydridic C-H bonds, even when weaker protic C-H bonds are present. This study describes the discovery and evolution of highly stable protoglobin nitrene transferases that catalyze the enantioselective intermolecular amination of the α-C-H bonds in carboxylic acid esters. We developed a high-throughput assay to evaluate the activity and enantioselectivity of mutant enzymes together with their sequences using the Every Variant Sequencing (evSeq) method. The assay enabled the identification of enantiodivergent enzymes that function at ambient conditions in Escherichia coli whole cells and whose activities can be enhanced by directed evolution for the amination of a range of substrates.

Biodegradation is recognized as the main route for the decomposition of phthalic acid esters (PAEs) in nature, but the fate of PAEs in marine ecosystems is not well understood. Herein, a novel marine bacterium, Gordonia sihwaniensis RL-BY03, was identified and analyzed for its ability to degrade PAEs. Furthermore, the metabolic mechanism of di-(2-ethylhexyl) phthalate (DEHP) was examined through UPLC-MS/MS and genomic analysis. RL-BY03 could rely solely on several types of PAEs as its sole carbon source. Initial pH and temperature for DEHP degradation were optimized as 8.0 and 30 °C, respectively. Surprisingly, RL-BY03 could simultaneously degrade ethyl acetate and DEHP and they could increase the cell surface hydrophobicity. DEHP degradation kinetics fitted well with the first-order decay model. The metabolic pathway of DEHP was deduced following the detection of five metabolic intermediates. Further, genes that are related to DEHP degradation were identified through genomic analysis and their expression levels were validated through RT-qPCR. A co-related metabolic pathway at biochemical and molecular level indicated that DEHP was turned into DBP and DEP by β-oxidation, which was further hydrolyzed into phthalic acid. Phthalic acid was utilized through catechol branch of β-ketoadipate pathway. Additionally, RL-BY03 exhibited excellent bioremediation potential for DEHP-contaminated marine samples. In general, these findings have the potential to enhance our understanding of the fate of PAEs in marine ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Phthalic acid esters (PAEs) are synthetic diesters derived from o-phthalic acid, commonly used as plasticizers. These compounds pose significant environmental and health risks due to their ability to leach into the environment and act as endocrine disruptors, carcinogens, and mutagens. Consequently, PAEs are now considered major emerging contaminants and priority pollutants. Microbial degradation, primarily by bacteria and fungi, offers a promising method for PAEs bioremediation. This article highlights the current state of microbial PAEs degradation, focusing on the major bottlenecks and associated challenges. These include the identification of novel and more efficient PAE hydrolases to address the complexity of PAE mixtures in the environment, understanding PAEs uptake mechanisms, characterizing novel o-phthalate degradation pathways, and studying the regulatory network that controls the expression of PAE degradation genes. Future research directions include mitigating the impact of PAEs on health and ecosystems, developing biosensors for monitoring and measuring bioavailable PAEs concentrations, and valorizing these residues into other products of industrial interest, among others., (© 2024 The Author(s). Microbial Biotechnology published by John Wiley & Sons Ltd.)

This study provides a comprehensive investigation into the structure-dependent uptake, distribution, biotransformation, and potential toxicity effects of alkyl organophosphate esters (OPEs) in hydroponic lettuce ( Lactuca sativa L.). Trimethyl, triethyl, and tripropyl phosphates were readily absorbed and acropetally translocated, while tributyl, tripentyl, and trihexyl phosphates accumulated mainly in lateral roots. The acropetal translocation potential was negatively associated with log K ow values. Trimethyl and triethyl phosphates are less prone to biotransformation, while a total of 14 novel hydrolysis, hydroxylated, and conjugated metabolites were identified for other OPEs using nontarget analysis. The extent of hydroxylation decreases from tripropyl phosphate to trihexyl phosphate, but multiple hydroxylations occurred more frequently on longer chain OPEs. Further comparative toxicity test revealed that hydrolyzed and hydroxylated metabolites have stronger toxic effects on Ca 2+ -dependent protein kinases (CDPK) than their parent OPEs. Dibutyl 3-hydroxybutyl phosphate particularly induces upregulation of CDPK in lateral roots of lettuce, probably associated with adenine reduction that may play an important role in the self-defense and detoxification processes. This study contributes to understanding the uptake and transformation behaviors of alkyl OPEs as well as their associations with a toxic effect on lettuce. This emphasizes the necessary evaluation of the environmental risk of the use of OPEs, particularly focusing on their hydroxylated metabolites.

Microalgal oil production represents a promising renewable biofuel source. Metabolic engineering can enhance its utility, transforming it into an improved biofuel and expanding its applications as a feedstock for commodity chemicals, thereby increasing their value in biorefineries. This study focused on anaerobic wax ester production by the microalga Euglena gracilis, aiming to develop stable mutant strains with altered wax ester profiles through genome editing. Two enzymes in the fatty acid beta-oxidation pathway involved in wax ester production were targeted-3-ketoacyl-CoA thiolase and acyl-CoA dehydrogenase-using clustered regularly interspaced short palindromic repeats/Cas9. The results revealed one genetic mutation that lengthened and three that shortened the distribution of wax ester compositions compared to the wild-type (WT). The triple-knockout mutant, combining mutations that shorten wax ester chains, produced wax esters with acyl chains two carbons shorter than WT. This study established a methodology to stably modify wax ester composition in E. gracilis., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Dienelactone hydrolase (DLH) is one of numerous hydrolytic enzymes with an α/β-hydrolase fold, which catalyze the hydrolysis of dienelactone to maleylacetate. The DLHs share remarkably similar tertiary structures and a conserved arrangement of catalytic residues. This study presents the crystal structure and comprehensive functional characterization of a novel thermostable DLH from the bacterium Hydrogenobacter thermophilus (HtDLH). The crystal structure of the HtDLH, solved at a resolution of about 1.67 Å, exhibits a canonical α/β-hydrolase fold formed by eight β-sheet strands in the core, with one buried α-helix and six others exposed to the solvent. The structure also confirmed the conserved catalytic triad of DHLs formed by Cys121, Asp170, and His202 residues. The HtDLH forms stable homodimers in solution. Functional studies showed that HtDLH has the expected esterase activity over esters with short carbon chains, such as p-nitrophenyl acetate, reaching optimal activity at pH 7.5 and 70 °C. Furthermore, HtDLH maintains more than 50 % of its activity even after incubation at 90 °C for 16 h. Interestingly, HtDLH exhibits catalytic activity towards polyethylene terephthalate (PET) monomers, including bis-1,2-hydroxyethyl terephthalate (BHET) and 1-(2-hydroxyethyl) 4-methyl terephthalate, as well as other aliphatic and aromatic esters. These findings associated with the lack of activity on amorphous PET indicate that HtDLH has characteristic of a BHET-degrading enzyme. This work expands our understanding of enzyme families involved in PET degradation, providing novel insights for plastic biorecycling through protein engineering, which could lead to eco-friendly solutions to reduce the accumulation of plastic in landfills and natural environments., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Extracellular hydrolysis of the phosphate esters of B vitamins (B1, B2, and B6) is crucial for their cellular uptake and metabolism. Although a few zinc-dependent enzymes have been implicated in these processes, their exact mechanisms of action remain largely unknown. This study investigated the potential involvement of phosphate group hydrolyzing enzymes in the hydrolysis of B vitamin phosphate esters. We evaluated enzyme activity in membrane lysates prepared from cells transiently transfected with these enzymes or those endogenously expressing them. Specifically, we investigated how zinc deficiency affects the rate of hydrolysis of B vitamin phosphate esters in cellular lysates. Assessment of the activities of zinc-dependent ectoenzymes in the lysates prepared from cells cultured in zinc-deficient conditions and in the serum of rats fed zinc-deficient diets revealed that zinc deficiency reduced the extracellular hydrolysis activity of B vitamin phosphate esters. Furthermore, our findings explain the similarities between several symptoms of B vitamin and zinc deficiencies. Collectively, this study provides novel insights into the diverse symptoms of zinc deficiency and could guide the development of appropriate clinical strategies., (© 2024 The Author(s). The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology.)

Sorghum, a globally grown gluten-free cereal, is used mainly as an animal feed in developed countries regardless of its potential for human consumption. In this study, we utilized nontargeted lipidomics to thoroughly analyze, compare, and characterize whole-grain lipids in six sorghum cultivars (cv) grown in a single field trial in Australia: Buster, Bazley, Cracker, Liberty, MR43, and Tiger. In total, 194 lipid molecular species representing five major lipid classes were identified. Multivariate analysis unveiled distinct lipid profiles among the cultivars. The most distinct lipid profile belonged to cv. MR43. The lower ω-6 to ω-3 ratio and optimal P/S ratio in cv. Bazley reflect this as a valuable source of balanced essential fatty acids in the diet. The novel bioactive lipids known as FAHFAs (fatty acid esters of hydroxy fatty acids) were identified and characterized in sorghum grains. These findings further emphasize the potential of whole-grain sorghum as a basis for new health-promoting food products.

Elevation of the plasma levels of ( S )-lactate (Lac) and/or ( R )-beta-hydroxybutyrate (BHB) occurs naturally in response to strenuous exercise and prolonged fasting, respectively, resulting in millimolar concentrations of these two metabolites. It is increasingly appreciated that Lac and BHB have wide-ranging beneficial physiological effects, suggesting that novel nutritional solutions, compatible with high-level and/or sustained consumption, which allow direct control of plasma levels of Lac and BHB, are of strong interest. In this study, we present a molecular hybrid between ( S )-lactate and the BHB-precursor ( R )-1,3-butanediol in the form of a simple ester referred to as LaKe . We show that LaKe can be readily prepared on the kilogram scale and undergoes rapid hydrolytic conversion under a variety of physiological conditions to release its two constituents. Oral ingestion of LaKe , in rats, resulted in dose-dependent elevation of plasma levels of Lac and BHB triggering expected physiological responses such as reduced lipolysis and elevation of the appetite-suppressing compound N -L-lactoyl-phenylalanine (Lac-Phe).

A new Candida parapsilosis ACCC 20221 ( C. parapsilosis ACCC 20221) whole-cell catalyst with a high phenolic glycoside esters synthesis activity and large biomass was obtained after culture with glucose. The possible mechanisms were revealed by using comparative proteomics. It found the expression of proteins involved in post-translational modification, protein turnover, and chaperone, and RNA processing and modification was upregulated, which ensured the metabolic balance and accurate translation, correct folding, and post-translational modification of proteins, thus enhancing the production of lipases in C. parapsilosis ACCC 20221 cultured with glucose. Moreover, the glycolysis pathway, tricarboxylic acid cycle, and fatty acids synthesis were enhanced, while the β-oxidation of fatty acids was weakened in C. parapsilosis ACCC 20221 cells cultured with glucose, which led to an increase in energy generation and cell membrane synthesis; thus, large biomass was obtained. In addition, CCE40476.1 and CAC86400.1, which were likely to exert arbutin esters synthesis activity in C. parapsilosis ACCC 20221, were screened, and it was found that vinyl propionate could easily enter the catalytic pocket of CCE40476.1 and form hydrogen bonding interactions with Leu191 and Ser266.

Bioaromas can be produced by lipases either through their hydrolytic or (trans)esterifying activities. Therefore, this work reports the development of a lipase-catalyzed biotransformed licuri oil, forming volatile ethyl esters with odor notes resembling tropical fruits. Ethyl octanoate formation was promoted when 7.0 % (m/v) Lipozyme 435® was used to convert a grain alcohol:licuri oil mixture (51:49, v/v) at 58ºC and 70 rpm for 6 hours. The biotransformed oil has shown antimicrobial activity against Staphylococcus hominis, S. epidermidis, and Corynebacterium xerosis, bacteria associated with bad skin odor. Finally, this biotransformed oil was used without further treatments (e.g., recovery or purification procedures) to prepare two cosmetic formulations (in a dosage of 1.5 %), aiming for both fragrant and deodorant activity., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: The authors declare that they have a patent application (BR102024007414–9) related to the results presented in this document., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study investigated the uptake pathways, acropetal translocation, subcellular distribution, and biotransformation of OPEs by rice ( Oryza sativa L.) after Cu exposure. The symplastic pathway was noted as the major pathway for the uptake of organophosphate triesters (tri-OPEs) and diesters (di-OPEs) by rice roots. Cu exposure enhanced the accumulation of tri-OPEs in rice roots, and such enhancement was positively correlated with Cu concentrations, attributing to the Cu-induced root damage. The hydrophilic Cl-OPEs in the cell-soluble fraction of rice tissues were enhanced after Cu exposure, while the subcellular distributions of alkyl- and aryl-OPEs were not affected by Cu exposure. Significantly higher biotransformation rates of tri-OPEs to di-OPEs occurred in leaves, followed by those in stems and roots. Our study reveals the mechanisms associated with the uptake, translocation, and biotransformation of various OPEs in rice after Cu exposure, which provides new insights regarding the phytoremediation of soils cocontaminated with heavy metal and OPEs.

Phthalic acid esters (PAE) are widely used as plasticizers and have been classified as ubiquitous environmental contaminants of primary concern. PAE have accumulated intensively in surface water, groundwater, and wastewaters; thus, PAE degradation is essential. In the present study, the ability of a saline soil bacteria (SSB)-consortium to degrade synthetic wastewater-phthalates with alkyl chains of different lengths, such as diethyl phthalate (DEP), di-n-butyl phthalate (DBP), benzyl butyl phthalate (BBP), and di (2-ethylhexyl) phthalate (DEHP) was characterized. A central composite design-response surface methodology was applied to optimize the degradation of each phthalate, where the independent variables were temperature (21-41 °C), pH (5.3-8.6) and PAE concentration (79.5-920.4 mg L -1 ), and Gas Chromatography-Mass Spectrometry was used to identify the metabolites generated during phthalate degradation. Optimal conditions were 31 °C, pH 7.0, and an initial PAE concentration of 500 mg L -1 , where the SSB-consortium removed 84.9%, 98.47%, 99.09% and 98.25% of initial DEP, DBP, BBP, and DEHP, respectively, in 168h. A first-order kinetic model explained - the biodegradation progression, while the half-life of PAE degradation ranged from 12.8 to 29.8 h. Genera distribution of the SSB-consortium was determined by bacterial meta-taxonomic analysis. Serratia, Methylobacillus, Acrhomobacter, and Pseudomonas were the predominant genera; however, the type of phthalate directly affected their distribution. Scanning electron microscopy analysis showed that high concentrations (1000 mg L -1 ) of phthalates induced morphological alterations in the bacterial SSB-consortium. The metabolite profiling showed that DEP, DBP, BBP, and DEHP could be fully metabolized through the de-esterification and β-oxidation pathways. Therefore, the SSB-consortium can be considered a potential candidate for bioremediation of complex phthalate-contaminated water resources., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:MARCO ANTONIO PEREYRA CAMACHO reports financial support was provided as fellowship by CONAHCyT no. 331852. ANTONIO DE LEON RODRIGUEZ reports financial support was provided by CONAHCyT Problemas Nacionales Grant no. 4601. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Despite being present in trace amounts, ethyl esters play a crucial role as flavour compounds in lager beer. In yeast, ethyl hexanoate, ethyl octanoate and ethyl decanoate, responsible for fruity and floral taste tones, are synthesized from the toxic medium chain acyl-CoA intermediates released by the fatty acid synthase complex during the fatty acid biosynthesis, as a protective mechanism. The aim of this study was to enhance the production of ethyl esters in the hybrid lager brewing yeast Saccharomyces pastorianus by improving the medium chain acyl-CoA precursor supply. Through CRISPR-Cas9-based genetic engineering, specific FAS1 and FAS2 genes harbouring mutations in domains of the fatty acid synthesis complex were overexpressed in a single and combinatorial approach. These mutations in the ScFAS genes led to specific overproduction of the respective ethyl esters: overexpression of ScFAS1 I306A and ScFAS2 G1250S significantly improved ethyl hexanoate production and ScFAS1 R1834K boosted the ethyl octanoate production. Combinations of ScFAS1 mutant genes with ScFAS2 G1250S greatly enhanced predictably the final ethyl ester concentrations in cultures grown on full malt wort, but also resulted in increased levels of free medium chain fatty acids causing alterations in flavour profiles. Finally, the elevated medium chain fatty acid pool was directed towards the ethyl esters by overexpressing the esterase ScEEB1. The genetically modified S. pastorianus strains were utilized in lager beer production, and the resulting beverage exhibited significantly altered flavour profiles, thereby greatly expanding the possibilities of the flavour palette of lager beers., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Wickerhamomyces anomalus is one of the most important ester-producing strains in Chinese baijiu brewing. Ethanol and lactic acid are the main metabolites produced during baijiu brewing, but their synergistic influence on the growth and ester production of W. anomalus is unclear. Therefore, in this paper, based on the contents of ethanol and lactic acid during Te-flavor baijiu brewing, the effects of different ethanol concentrations (3, 6, and 9% (v/v)) combined with 1% lactic acid on the growth and ester production of W. anomalus NCUF307.1 were studied and their influence mechanisms were analyzed by transcriptomics. The results showed that the growth of W. anomalus NCUF307.1 under the induction of lactic acid was inhibited by ethanol. Although self-repair mechanism of W. anomalus NCUF307.1 induced by lactic acid was initiated at all concentrations of ethanol, resulting in significant up-regulation of genes related to the Genetic Information Processing pathway, such as cell cycle-yeast, meiosis-yeast, DNA replication and other pathways. However, the accumulation of reactive oxygen species and the inhibition of pathways associated with carbohydrate and amino acid metabolism may be the main reason for the inhibition of growth in W. anomalus NCUF307.1. In addition, 3% and 6% ethanol combined with 1% lactic acid could promote the ester production of W. anomalus NCUF307.1, which may be related to the up-regulation of EAT1, ADH5 and TGL5 genes, while the inhibition in 9% ethanol may be related to down-regulation of ATF2, EAT1, ADH2, ADH5, and TGL3 genes., Competing Interests: Declaration of competing interest The manuscript entitled “Transcriptomics to evaluate the influence mechanisms of ethanol on the ester production of Wickerhamomyces anomalus with the induction of lactic acid” is submitted for your consideration for publication as a communication in “Food Microbiology”. No conflict of interest exists in the submission of this manuscript, and the manuscript is approved by all authors for publication. I would like to declare on behalf of my co-authors that the work described was original research that has not been published previously and is not under consideration for publication elsewhere, in whole or in part., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

As the substitutes of polybrominated diphenyl ethers, organophosphate esters (OPEs) with high concentrations have accumulated in the estuaries, bays, and harbors. However, limited information is available about the OPEs in the estuary organism categories, especially under the multiple industrial pressure. This study investigated the occurrence, bioaccumulation and human consumption implication in wild marine organisms from the Yellow River Estuary, where located many petroleum and chemical manufacturing industries. This study found that concentrations of Σ 13 OPEs ranged from 547 ng/L to 1164 ng/L in seawater (median: 802 ng/L), from 384 to 1366 ng/g dw in the sediment (median: 601 ng/g dw), and from 419 to 959 ng/g dw (median: 560 ng/g dw) in the marine organisms. The congener compositions in the organisms were dominated by alkyl-OPEs (80.7 %), followed by halogenated-OPEs (18.8 %) and aryl-OPEs (0.5 %). Based on the principal component analysis, petrochemical pollution, and industrial wastewater discharge were distinguished as the main plausible sources of OPEs to the YRE ecosystem. Most OPEs had potential or strong bioaccumulation capacity on the organisms, with a positive correlation between log BAF (Bioaccumulation Factor) and log Kow of OPEs. The highest estimated daily intake value of OPEs was tri-n-propyl phosphate, exceeding 300 ng/kg·bw/day via consuming fish. The highest hazard quotients from OPEs ranged from 0.001 to 0.1, indicating a low risk to human health by consuming marine organisms in the YRE. As the consumption of OPEs increases year by year, the risks of OPEs still cannot be ignored., Competing Interests: Declaration of competing interest All authors of this article declare that they have no conflict of interest regarding the publication of this study., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Food waste (FW) and its biogas residue were considered as sources of terrestrial microplastics (MPs) and phthalic acid esters (PAEs) contamination. However, there was a lack of research and understanding of the MPs and PAEs pollution problem in FW dry anaerobic digestion process (DADP). The MPs and PAEs in three stages of the DADP with the largest monomer disposal scale in China were identified. At the biogas residue extrusion stage, MPs abundance and PAEs concentration reached the highest values, which were 3.63 ± 0.45 × 10 3 N·kg -1 and 3.62 ± 0.72 mg·kg -1 , respectively. Furthermore, there was a significant positive correlation between MPs and PAEs throughout the process (p < 0.05). Although bacteria and fungi with plastic degradation potential were present in all stages, the contamination problem of MPs and PAEs cannot be completely solved through DADP. This study provides a scientific basis for preventing and controlling the pollution of MPs and PAEs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Untargeted metabolomics is a powerful profiling tool for the discovery of possible biomarkers of disease onset and progression. Analytical pipelines applying liquid chromatography (LC) and mass spectrometry (MS)-based methods are widely used to survey a broad range of metabolites within various metabolic pathways, including organic acids, amino acids, nucleosides, and lipids. Accurate and complete identification of putative metabolites is an ongoing challenge in untargeted metabolomics studies. Highly sensitive instrumentation can result in the detection of adduct and fragment ions that form reproducibly and contain identifiable ions that are difficult to distinguish from metabolic pathway intermediates, which may result in false-positive identification. At concentrations as low as 10 μM, free fatty acids have been found to form homo- and heterodimers in untargeted metabolomics pipelines that resemble the lipid class fatty acid esters of hydroxy fatty acids (FAHFAs), resulting in misidentification. This chapter details a protocol for LC-MS-based untargeted metabolomics using hydrophilic interaction chromatography (HILIC) that specifically aids in distinguishing artifactual fatty acid dimers from endogenous FAHFAs., (© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

This study explores the behavior of organophosphate esters (OPEs) in different species within marine ecosystems and their potential for bioaccumulation and biomagnification. The concentrations of OPEs were analyzed in marine species (krill (Meganyctiphanes norvegica), jellyfish (Pelagia noctiluca), European sardine (Sardina pilchardus), European anchovy (Engraulis encrasicolus), European hake (Merluccius merluccius), loggerhead turtle (Caretta caretta), European squid (Loligo vulgaris), fin whale (Balaenoptera physalus) and striped dolphin (Stenella coeruleoalba)) from different trophic levels, to understand their distribution and contamination profiles. The study provides insights into the metabolism of OPEs and their biomagnification on species occupying higher trophic levels. The results show that the differences in OPE concentrations among species are influenced by contamination levels at sampling points, as well as species and trophic level characteristics. The study reveals that the sum of OPEs do not exhibit significant biomagnification within the marine food web, with higher trophic level species showing efficient metabolism of these contaminants. However, biomagnification analysis of individual compounds demonstrates that some OPEs, such as tris(2-ethylhexyl) phosphate (TEP), tris(2-butoxyethyl) phosphate (TBOEP), and tris(2-isopropylphenyl) phosphate (T2IPPP) consistently exhibit biomagnification within marine trophic webs, while other show different behaviors depending on the trophic web. The loggerhead turtle shows higher biomagnification for specific OPEs, indicating the influence of diet and direct plastic waste consumption. Furthermore, OPEs prone to metabolism, such as tri-n-butyl phosphate (TNBP) and TBOEP, are present in multiple species across different trophic levels, suggesting a tendency for bioaccumulation. The study highlights the complexity of OPE behavior and the need to evaluate the biomagnification potential of each compound individually. It also emphasizes the toxic effects associated with OPE exposure and the potential risks to organisms within marine ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Phytosterols are plant sterols that are important secondary plant metabolites with significant pharmacological properties. Their presence in the plant kingdom concerns many unrelated botanical families such as oleageneous plants and cereals. The structures of phytosterols evoke those of cholesterol. These molecules are composed of a sterane ring, also known as perhydrocyclopentanophenanthrene, along with a methyl or ethyl group at C-24 in their side chains, a hydroxyl group at C-3 on ring A, and one or two double bonds in the B ring. Phytosterols display different oxidation degrees at the sterane ring and at the side chain as well as varying numbers of carbons with complex stereochemistries. Fats and water solubilities of phytosterols have been achieved by physical, chemical and enzymatic esterifications to favor their bioavailability and to improve the sensory quality of food, and the efficiency of pharmaceutic and cosmetic products. This review aims to provide comprehensive information starting from the definition and structural classification of phytosterols, and exposes an update of their biogenic relationships. Next, the synthesis of phytosterol esters and their applications as well as their effective roles as hormone precursors are discussed. Finally, a concise exploration of the latest advancements in phytosterol / oxyphytosterols analysis techniques is provided, with a particular focus on modern hyphenated techniques., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Crocetin di/mono-glucosyl esters (crocin-4 and crocin-5) are rarely distributed in nature, limiting their potential applications in the food and pharmaceutical industries. In the present study, a novel GH3 family β-glucosidase Lf18920 was identified from Leifsonia sp. ZF2019, which selectively hydrolyzed crocin-1 (crocetin di-gentiobiosyl ester) to crocin-5 and crocin-4, but not to its aglycone, crocetin. Under the optimal condition of 40 °C and pH 6.0 for 120 min, Lf18920 almost completely hydrolyzed crocin-1, yielding 73.50±5.66 % crocin-4 and 16.19±1.38 % crocin-5. Molecular docking and point mutation studies revealed that Lf18920 formed a narrow binding channel that facilitated crocin-1 binding. Five single amino acid variants (D50A, D53A, W274A, G420A, and Q421A) were constructed, all of which showed reduced hydrolytic activity. Mutations at D50 and D53, located distal to the active site, increased binding energy and decreased hydrolytic activity, while mutations at W274, G420, and Q421, proximal to the active site, disrupted hydrolytic function. These findings suggest that the narrow binding channel and specific enzyme-substrate interactions are crucial for Lf18920's selective hydrolytic activity. Overall, this study is the first to report a β-glucosidase capable of selectively transforming crocin-1 to crocetin di/mono-glucosyl esters, offering potential for synthesizing crocin-4 and crocin-5., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

With the wide application of phthalic acid esters (PAEs) in the manufacturing of plastic products, a large number of PAEs were discharged into marine ecosystem and accumulated in fish, which has posed a serious threat to marine ecological environment and fishery resources. However, the bioaccumulation of PAEs in fish in mangrove ecosystem, the most productive marine ecosystem, has not been well characterized. In this study, dominant fish and their potential food sources (including particulate organic matter (POM), sedimentary organic matter (SOM), Metapenaeus ensis (Shrimp) and Oreochromis (Ore) were collected from Dongzhai Harbor, a typical mangrove ecosystem. The concentrations of nine PAEs in fish and their potential food sources were determined. Then stable nitrogen and carbon isotope analysis, combined with a new Bayesian mixing model (MixSIMMR) was used to quantify the diet compositions of fish and elucidate the effect of dietary habit on PAEs bioaccumulation in fish. The results indicated that the median concentration of ∑ 9 PAEs in fish was 1119 μg/kg ww, positioning it at a moderate to low level in comparison to other regions. di-n-butyl phthalate (DBP) and diisononyl ortho-phthalate (DINP) were the dominant PAEs in fish. The PAEs concentration in demersal fish was significantly higher than that of pelagic fish, which may be attributed to the substantial contributions of shrimp (28.5 %) and POM (25.3 %) to the diet of demersal fish. This study provided new insights on the bioaccumulation of PAEs in dominant mangrove fish and confirmed that habitat preferences and food sources could significantly influence the bioaccumulation of PAEs in fish., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Fatty acid esters of hydroxy fatty acids (FAHFAs) are endogenous bioactive lipids known for their anti-inflammatory and anti-diabetic properties. Despite their therapeutic potential, little is known about the sex-specific variations in FAHFA metabolism. This study investigated the role of sex and Androgen Dependent TFPI Regulating Protein (ADTRP), a FAHFA hydrolase. Additionally, tissue-specific differences in FAHFA levels, focusing on the perigonadal white adipose tissue (pgWAT), subcutaneous white adipose tissue (scWAT), brown adipose tissue (BAT), plasma, and liver, were evaluated using metabolomics and lipidomics. We found that female mice exhibited higher FAHFA levels in pgWAT, scWAT, and BAT compared to males. FAHFA levels were inversely related to testosterone and Adtrp mRNA, which showed significantly lower expression in females compared with males in pgWAT and scWAT. However, no significant differences between the sexes were observed in plasma and liver FAHFA levels. Adtrp deletion had minimal impact on both sexes' metabolome and lipidome of pgWAT. However, we discovered higher endogenous levels of triacylglycerol estolides containing FAHFAs, a FAHFA metabolic reservoir, in the pgWAT of female mice. These findings suggest that sex-dependent differences in FAHFA levels occur primarily in specific WAT depots and may modulate local insulin sensitivity in adipocytes, and the role of ADTRP is limited to adipose depots. However, further investigations are warranted to fully comprehend the underlying mechanisms and implications of sex-dependent regulation of human FAHFA metabolism., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Most Phalaenopsis cultivars have almost no aroma, with a few exceptions. Phalaenopsis presents significant challenges in fragrance breeding due to its weak aroma and low fertility. It is therefore necessary to identify the aroma components and key regulatory genes in Phalaenopsis cultivars like 'Orange Beauty', 'Brother Sara Gold', 'Purple Martin', 'H026', 'SK16', 'SX098', and 'SH51', to improve the aroma of the common Phalaenopsis., Results: Floral aroma components were tested on nine Phalaenopsis species, using smell identification and headspace gas chromatography-mass spectrometry. The result showed that alcohols, esters, and alkenes were the key specific components in the different species and cultivar aromas and the aroma intensity and component content of cultivars with different colors were different. The main components of the floral aromas in Phalaenopsis were alcohols (including eucalyptol, linalool, citronellol, and 1-hexanol), esters (including hexyl acetate, leaf acetate, and dibutyl phthalate), alkenes (including pinene and sabinene) and arenes (like fluorene). The transcriptome of flowers in the bud stage and bloom stage of P. 'SH51' was sequenced and 5999 differentially expressed genes were obtained. The contributions of the phenylpropionic acid/phenyl ring compound and the terpene compound to the aroma were greater. Sixteen genes related to phalaenopsis aroma were found. TC4M, PAL, CAD6, and HR were related to phenylpropanoid synthesis pathway. SLS, TS10, and P450 were related to the synthesis pathway of terpenes. TS10 and YUCCA 10 were involved in tryptophan metabolism., Conclusion: This is the first report on the floral aroma components and regulatory genes in Phalaenopsis. The proposed method and research data can provide technical support for Phalaenopsis breeding. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Phenylpropanoid sucrose esters are a large and important group of natural substances with significant therapeutic potential. This work describes a pilot study of the enzymatic hydroxycinnamoylation of sucrose and its derivatives which was carried out with the aim of obtaining precursors of natural phenylpropanoid sucrose esters, e.g., vanicoside B. In addition to sucrose, some chemically prepared sucrose acetonides and substituted 3'- O -cinnamates were subjected to enzymatic transesterification with vinyl esters of coumaric, ferulic and 3,4,5-trimethoxycinnamic acid. Commercial enzyme preparations of Lipozyme TL IM lipase and Pentopan 500 BG exhibiting feruloyl esterase activity were tested as biocatalysts in these reactions. The substrate specificity of the used biocatalysts for the donor and acceptor as well as the regioselectivity of the reactions were evaluated and discussed. Surprisingly, Lipozyme TL IM catalyzed the cinnamoylation of sucrose derivatives more to the 1'-OH and 4'-OH positions than to the 6'-OH when the 3'-OH was free and the 6-OH was blocked by isopropylidene. In this case, Pentopan reacted comparably to 1'-OH and 6'-OH positions. If sucrose 3'- O -coumarate was used as an acceptor, in the case of feruloylation with Lipozyme in CH 3 CN, 6- O -ferulate was the main product (63%). Pentopan feruloylated sucrose 3'- O -coumarate comparably well at the 6-OH and 6'-OH positions (77%). When a proton-donor solvent was used, migration of the 3'- O -cinnamoyl group from fructose to the 2-OH position of glucose was observed. The enzyme hydroxycinnamoylations studied can shorten the targeted syntheses of various phenylpropanoid sucrose esters.

Lipophenols, phenolic compounds esterified with fatty alcohols or fatty acids, provide greater health benefits upon dietary ingestion of plant-based foods than unesterified (poly)phenols. Based on this premise, the present study aimed to demonstrate the role of gastrointestinal enzymes (pepsin, pancreatin, and pancreatic lipase) in releasing alkyl gallates and trans -caffeates from wine lees, providing bioactive compounds with enhanced capacities against oxidative stress (OS) and para-inflammation. The UHPLC-ESI-QqQ-MS/MS-based analysis revealed ethyl gallate and ethyl trans -caffeate as the most prominent compounds (1.675 and 0.872 μg/g dw, respectively), while the bioaccessibility of the derivatives of gallic and caffeic acids was dependent on the alkyl chain properties. The de novo formation of alkyl gallates during gastric and intestinal digestion resulted from intestinal enzyme activity. Moreover, the in vitro capacity of bioaccessible alkyl esters of gallic and trans -caffeic acids to reduce cyclooxygenase-2 concentration and modulate oxilipins related to OS (8-iso-PGF 2α ) and inflammation (PGF 2α and PGE 2 ) was demonstrated in a time-dependent manner. In conclusion, the presence of alkyl esters of gallic and trans -caffeic acids in wine lees and their subsequent formation during digestion of this byproduct emphasize their value as a source of antioxidant and anti-inflammatory compounds, encouraging the consideration of wine lees as a valuable ingredient for health-promoting coproducts.

Asperalins represent a novel class of viridicatin natural products with potent inhibitory activities against fish pathogens. In this study, we elucidated the biosynthesis of asperalins in the Aspergillus oryzae NSAR1 heterologous host and identified the FAD-dependent monooxygenase AplB stereoselectively hydroxylates viridicatin to yield a unique 3 R ,4 S configuration. The monomodular NRPS AplJ catalyzes a rare intramolecular ester bond formation reaction using dihydroquinoline as a nucleophile. Subsequent modifications by cytochrome P450 AplF, chlorinase AplN, and prenyltransferase AplE tailor the anthranilic acid portion, leading to the formation of asperalins. Additionally, we explored the potential of AplB for the hydroxylation of viridicatin analogs, demonstrating its relaxed substrate specificity. This finding suggests that AplB could be developed as a biocatalyst for the synthesis of viridicatin derivatives.

Phthalate esters (PAEs) are widely used as plasticizers and cause serious complex pollution problem in environment. Thus, strains with efficient ability to simultaneously degrade various PAEs are required. In this study, a newly isolated strain Rhodococcus sp. AH-ZY2 can degrade 500 mg/L Di-n-octyl phthalate completely within 16 h and other 500 mg/L PAEs almost completely within 48 h at 37 °C, 180 rpm, and 2 % (v/v) inoculum size of cultures with a OD 600 of 0.8. OD 600 = 0.8, 2 % (v/v). Twenty genes in its genome were annotated as potential esterase and four of them (3963, 4547, 5294 and 5359) were heterogeneously expressed and characterized. Esterase 3963 and 4547 is a type I PAEs esterase that hydrolyzes PAEs to phthalate monoesters. Esterase 5294 is a type II PAEs esterase that hydrolyzes phthalate monoesters to phthalate acid (PA). Esterase 5359 is a type III PAEs esterase that simultaneously degrades various PAEs to PA. Molecular docking results of 5359 suggested that the size and indiscriminate binding feature of spacious substrate binding pocket may contribute to its substrate versatility. AH-ZY2 is a potential strain for efficient remediation of PAEs complex pollution in environment. It is first to report an esterase that can efficiently degrade mixed various PAEs., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have influenced the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Phthalate esters (PAEs), as a major plasticizer with multi-biotoxicity, are frequently detected in marine environments, and potentially affecting the survival of aquatic organisms. In the study, three typical PAEs (dimethyl phthalate [DMP], dibutyl phthalate [DBP] and di(2-ethylhexyl) phthalate [DEHP]) were selected to investigate the accumulation patterns and ecotoxicological effects on Mytilus coruscus (M. coruscus). In M. coruscus, the accumulation was DEHP>DBP>DMP, and the bioaccumulation in tissues was digestive glands>gills>gonads>muscles. Meanwhile, the activities of superoxide dismutase (SOD) and catalase (CAT) showed an activation-decrease-activation trend of stress, with more pronounced concentration effects. Glutathione reductase (GSH) activity was significantly increased, and its expression was more sensitive to be induced at an early stage. The metabolic profiles of the gonads, digestive glands and muscle tissues were significantly altered, and DEHP had a greater effect on the metabolic profiles of M. coruscus, with the strongest interference. PAEs stress for 7 d significantly altered the volatile components of M. coruscus, with potential implications for their nutritional value. This study provides a biochemical, metabolomic, and nutritional analysis of DMP, DBP, and DEHP toxic effects on M. coruscus from a multidimensional perspective, which provides support for ecotoxicological studies of PAEs on marine organisms. ENVIRONMENTAL IMPLICATION: Phthalate esters (PAEs), synthetic compounds from phthalic acid, are widespread in the environment, household products, aquatic plants, animals, and crops, posing a significant threat to human health. However, the majority of toxicological studies examining the effects of PAEs on aquatic organisms primarily focus on non-economic model organisms like algae and zebrafish. Relatively fewer studies have been conducted on marine organisms, particularly economically important shellfish. So, this study is innovative and necessary. This study provides a biochemical, metabolomic, and nutritional analysis of DMP, DBP, and DEHP toxic effects on mussels, and supports the ecotoxicology of PAEs on marine organisms., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Odorant-binding proteins (OBPs) initiate insect olfactory perception and mediate specific binding and selection of odorants via uncertain binding mechanisms. We characterized the binding characteristics of four OBPs from the striped flea beetle Phyllotreta striolata (SFB), a major cruciferous crop pest. Tissue expression analysis revealed that the two ABPII OBPs (PstrOBP12 and PstrOBP19) were highly expressed mainly in the antenna, whereas the two minus-C OBPs (PstrOBP13 and PstrOBP16) showed a broad expression pattern. Competitive binding assays of cruciferous plant volatiles showed that PstrOBP12, PstrOBP16 and PstrOBP19 had very strong binding capacities for only two phthalate esters (K i  < 20 μM), and PstrOBP13 specifically bound to four aromatic volatiles (K i  < 11 μM). Fluorescence quenching assays displayed that two phthalate esters bound to three PstrOBPs via different quenching mechanisms. PstrOBP12/PstrOBP16-diisobutyl phthalate and PstrOBP19-bis(6-methylheptyl) phthalate followed static quenching, while PstrOBP12/PstrOBP16-bis(6-methylheptyl) phthalate and PstrOBP19-diisobutyl phthalate followed dynamic quenching. Homology modelling and molecular docking displayed that PstrOBP12-diisobutyl phthalate was driven by H-bonding and van der Waals interactions, while PstrOBP16-diisobutyl phthalate and PstrOBP19-bis(6-methylheptyl) phthalate followed hydrophobic interactions. Finally, behavioural activity analysis demonstrated that phthalate esters exhibited different behavioural activities of SFB at different doses, with low doses attracting and high doses repelling. Overall, we thus revealed the different binding properties of the three PstrOBPs to two phthalate esters, which was beneficial in shedding light on the ligand-binding mechanisms of OBPs., (© 2024 Royal Entomological Society.)

A large number of studies on organophosphate esters (tri-OPEs) in marine organisms have not assessed the simultaneous occurrence of tri-OPEs and their metabolites (di-OPEs) in these species. This research investigated the concentration and geographical distribution of 15 tri-OPEs and 7 di-OPEs in 172 samples of Pampus argenteus that were collected annually from 2021 to 2023 at three distinct locations along the Vietnamese coast. As a result, tri-OPEs and di-OPEs were detected in numerous fish samples, indicating their widespread spatial and temporal occurrence in marine fish and pointing out the importance of monitoring their levels. The tri-OPEs and di-OPEs ranged within 2.1-38.9 ng g -1 dry weight (dw) and 3.2-263.4 ng g -1 dw, respectively. The mean concentrations of tri-OPEs ranged from 0.4 (TIPrP) to 5.4 ng g -1 dw (TBOEP), with TBOEP and TEHP having the highest mean values. In addition, the profiles of tri-OPEs in fish exhibited a descending order: Σalkyl OPEs > ΣCl-alkyl OPEs > Σaryl OPEs. The di-OPEs, namely BEHP and DMP, had the highest mean levels, measuring 33.4 ng g -1 dw and 23.8 ng g -1 dw, respectively. Furthermore, there have been significant findings of strong positive correlations between di-OPEs and tri-OPE pairs (p < 0.05). It is worth noting that there is a noticeable difference in the composition of tri-OPEs between the North and other regions. Despite these findings, the presence of OPE-contaminated fish did not pose any health risks to Vietnam's coastal population., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The concerning of plastic pollution in different ecosystems has been worsened by the widespread presence. Phthalate esters (PAEs), plasticizers found in everyday products, can migrate into the environment, especially into the oceans. Researches on their effects on cetaceans are still rare. Metabolomics helps assess perturbations induced by exposure to PAEs, which act as persistent endocrine disruptors. Four PAEs (dimethyl phthalate - DMP, diethyl phthalate - DEP, dibutyl phthalate - DBP, and di(2-ethylhexyl phthalate - DEHP) were analyzed, along with cholesterol and fatty acid profiles of P. blainvillei's blubber samples collected in southern Brazil. The study reveals pervasive contamination by PAEs - especially DEHP, present in all samples - with positive correlations between DEP content and animal size and weight, as well as between the DEHP amount and the C17:1 fatty acid. These findings will be relevant to conservation efforts aimed at this threatened species and overall marine ecosystems., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The cellular response to oxidants or xenobiotics comprises two key pathways, resulting in modulation of NRF2 and FOXO transcription factors, respectively. Both mount a cytoprotective response, and their activation relies on crucial protein thiol moieties. Using fumaric acid esters (FAEs), known thiol-reactive compounds, we tested for activation of NRF2 and FOXO pathways in cultured human hepatoma cells by dimethyl/diethyl as well as monomethyl/monoethyl fumarate. Whereas only the diesters caused acute glutathione depletion and activation of the stress kinase p38 MAPK , all four FAEs stimulated NRF2 stabilization and upregulation of NRF2 target genes. However, no significant FAE-induced activation of FOXO-dependent target gene expression was observed. Therefore, while both NRF2 and FOXO pathways are responsive to oxidants and xenobiotics, FAEs selectively activate NRF2 signaling., (© 2024 The Author(s). FEBS Open Bio published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)

Synthetic aromatic esters, widely employed in agriculture, food, and chemical industries, have become emerging environmental pollutants due to their strong hydrophobicity and poor bioavailability. This study attempted to address this issue by extracellularly expressing the promiscuous aminopeptidase (Aps) from Pseudomonas aeruginosa GF31 in B. subtilis , achieving an impressive enzyme activity of 13.7 U/mg. Notably, we have demonstrated, for the first time, the Aps-mediated degradation of diverse aromatic esters, including but not limited to pyrethroids, phthalates, and parabens. A biochemical characterization of Aps reveals its esterase properties and a broader spectrum of substrate profiles. The degradation rates of p-nitrobenzene esters (p-NB) with different side chain structures vary under the action of Aps, showing a preference for substrates with relatively longer alkyl side chains. The structure-dependent degradability aligns well with the binding energies between Aps and p-NB. Molecular docking and enzyme-substrate interaction elucidate that hydrogen bonding, hydrophobic interactions, and π-π stacking collectively stabilize the enzyme-substrate conformation, promoting substrate hydrolysis. These findings provide new insights into the enzymatic degradation of aromatic ester pollutants, laying a foundation for the further development and modification of promiscuous enzymes.

Phthalates esters (PAEs) are a kind of polymeric material additives widely been added into plastics to improve products' flexibility. It can easily cause environmental pollution which are hazards to public health. In this study, we isolated an efficient PAEs degrading strain, Janthinobacterium sp. E1, and determined its degradation effect of di-2-ethylhexyl phthalate (DEHP) under stress conditions. Strain E1 showed an obvious advantage in pollutants degradation under various environmental stress conditions. Degradation halo clearly occurred around the colony of strain E1 on agar plate supplemented with triglyceride. Strain E1's esterase is a constitutively expressed intracellular enzyme. The esterase purified from strain E1 showed a higher catalytic effect on short-chain PAEs than long-chain PAEs. The input of DEHP, DBP (dibutyl phthalate) and DMP (dimethyl phthalate) into the tested soil did not change the species composition of soil prokaryotic community, but altered the dominant species in specific environmental conditions. And the community diversity and richness decreased to a certain extent. However, the diversity and richness of the microbial community were improved after the contaminated soil was treated with the strain E1. Our results also suggested that strain E1 exhibited a tremendous potential in environmental bioremediation in the real environment, which provides a new insight into the elimination of the pollutants contamination in the urban environment.

To explore key factors involved in the uptake, translocation and accumulation of organophosphate esters (OPEs), computer simulation analysis and hydroponic experiments were executed. Lipid transporters with stocky-like active (SAC) cavities usually showed stronger binding affinities with the OPEs, especially when the SAC cavities belong to the Fish Trap model according to molecular docking. In our hydroponic trial, the binding affinity and gene expression of the lipid transporters and log  K ow of the OPEs could be charged to the uptake, translocation and accumulation of the OPEs; however, these three factors played various important roles in roots and shoots. In detail, the effect of gene expression and binding affinity were stronger than log  K ow in roots uptake and accumulation, but the result was the opposite in the shoots translocation. Transporters OsTIL and OsLTPL1 among all investigated transporters could play key roles in transporter-mediated OPE uptake, translocation and accumulation in the roots and shoots. OsMLP could be involved in the bidirected vertical translocation of the OPEs. OsLTP2 and OsLTP4 mainly acted as transporters of the OPEs in roots.

This study represents the initial examination of the herbicidal efficacy, crop safety, and degradation patterns of 2,4-D ethylhexyl ester (2,4-D EHE) at the enantiomeric level. Baseline separation of 2,4-D EHE enantiomers was achieved using a superchiral R-AD column, with their absolute configurations determined through chemical reaction techniques. Evaluation of weed control efficacy against sensitive species such as sun spurge and flixweed demonstrated significantly higher inhibition rates for S -2,4-D EHE compared to R -2,4-D EHE. Conversely, no stereoselectivity was observed in the fresh-weight inhibition rates of both enantiomers on crops or nonsensitive weeds. A sensitive HPLC-MS/MS method was developed to simultaneously detect two enantiomers and the metabolite 2,4-D in plants. Investigation into degradation kinetics revealed no substantial difference in the half-lives of R - and S -2,4-D EHE in maize and flixweed. Notably, the metabolite 2,4-D exhibited prolonged persistence at elevated levels on flixweed, while it degraded rapidly on maize.