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BACKGROUND: The present study explored the effects of pre‐harvest methyl jasmonates spraying on the volatiles of 'Cabernet Gernischt' grapes through the lipoxygenase pathway. Headspace solid‐phase microextraction combined with gas chromatography‐mass spectrometry and an enzyme‐linked immunosorbent assay were utilized to analyze volatile metabolites and key enzyme activities following methyl jasmonates application. Total RNA extraction and cDNA library construction were followed by transcriptome sequencing. RESULTS: The application of 0.1 mmol L−1 methyl jasmonate and 5 mmol L−1 methyl dihydrojasmonate significantly increased the levels of C6 compounds in 'Cabernet Gernischt' berries, and also enhanced the utilization of unsaturated fatty acids as precursors in the lipoxygenase‐hydroperoxides lyase pathway. The up‐regulated expression of VvADH1, VvADH2, VvHPL and VvLOX2S genes led to modulations in enzyme activity, thereby enhancing the berries's aromatic profile. There was a significant correlation between linoleic and linolenic acids (i.e. the precursors of the lipoxygenase pathway) and the activities and metabolites of key enzymes. CONCLUSION: The optimal concentration of methyl jasmonates treatment favorably influences the accumulation of green leaf volatiles in 'Cabernet Gernischt' grape. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

In this study, the optimal concentrations of methyl jasmonate (MeJA) and methyl dihydrojasmonate (MDJA) for Cabernet Gernischt were selected based on basic physicochemical properties of grapes. The key volatile metabolites and key enzyme activities in response to MeJA and MDJA were analyzed by headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) and enzyme-linked immunosorbent assay (ELISA), respectively. The effects of pre-harvest MeJA or MDJA treatment on the fruit quality and green leaf volatile components in the lipoxygenase (LOX) metabolic pathway of Cabernet Gernischt were also investigated. The findings revealed that the application of 0.1 mmol/L MeJA or 5 mmol/L MDJA significantly enhanced mass of 100 berries, total sugar content, soluble solids content, pH, total phenol and total anthocyanin contents. Compared with the control group, MeJA increased these indexes by 10.24%, 27.83%, 28.24%, 4.80%, 77.63% and 85.29%, respectively, while MDJA increased them by 9.15%, 24.01%, 24.41%, 8.30%, 69.21% and 71.57%, respectively. Furthermore, MeJA and MDJA reduced titratable acid content by 40.0% and 30.24%, respectively. Accordingly, both MeJA and MDJA significantly improved fruit quality. Moreover, they significantly increased the contents of C6 compounds responsible for the typical aroma of Cabernet Gernischet. Significant correlations were observed between the activity of each key enzyme in the lipoxygenase metabolic pathway and the corresponding metabolites, indicating that the application of MeJA or MDJA at a suitable concentration could positively influence the accumulation of green leaf volatiles in Cabernet Gernischt.

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The molecular docking was explored to study the interactions between reactive species generated by cold plasma and the enzymes lipoxygenase (LOX) and lipase (LPS), with the aim of elucidating the molecular mechanisms governing these interactions. Molecular docking results suggest that both LOX and LPS are primarily involved in hydrogen bonding interactions with the seven reactive species. The key binding sites for LOX and LPS were identified as Ile 663 and Glu 188, respectively. Notably, the lowest docking energy was observed between LOX and NO (-13.75 kcal/mol), whereas for LPS, it is between LPS and NO 3 (-12.08 kcal/mol). Increased treatment voltage and time resulted in higher inactivation levels, with LPS exhibiting higher residual activity compared to LOX. When the voltage was 75 kV and the time was 120 s, the residual activities of LOX and LPS were 42.88% and 56.77%, respectively. Consequently, the results enhance our understanding of the mechanisms underlying the inhibition of enzyme activity by reactive species generated by cold plasma. Moreover, cold plasma may serve as a novel preservation technology for inhibiting lipid oxidation of food by controlling enzyme activity., 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 quality deterioration of paddy during storage is closely associated to lipid metabolism. To explore the effect of lipid metabolism on the texture of paddy, freshly harvested Nanjingxiangzhan (Indica rice) stored for 60 days at 15 °C and 25 °C for 60 days was investigated. Paddy stored at 15 °C showed higher contents of ATP, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, triacylglycerols, diacylglycerols, monogalactosyldiacylglycerol and digalactosyldiacylglycerol but lower levels of lysophosphatidylcholine, lysophosphatidylethanolamine, phosphatidic acid, ceramides and free fatty acids. Storage at 15 °C inhibited lipase, phospholipase D (PLD), lipoxygenase (LOX) activities and the corresponding gene expressions. Moreover, 15 °C storage retarded the rise of hardness, cohesiveness and chewiness, while delayed the reduction of gumminess and springiness. These findings suggested that maintenance in glycerophospholipids, glycerolipids and saccharolipids abundance, reduction in lysophospholipids, phosphatidic acid, ceramides and free fatty acids accumulation could contribute to enhanced internal resistance to aging in freshly harvested paddy at low temperature storage., 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.)

Plants emit an array of volatile organic compounds in response to stresses. Six-carbon green leaf volatiles (GLVs) and five-carbon pentyl leaf volatiles (PLVs) are fatty acid-derived compounds involved in intra- and inter-species communications. Unlike extensively studied GLVs, the biological activities of PLVs remain understudied. Maize (Zea mays L.) contains a unique monocot-specific lipoxygenase, ZmLOX6, that is unable to oxidize fatty acids and instead possesses a hydroperoxide lyase-like activity to specifically produce PLVs. Here, we show that disruption of ZmLOX6 reduced resistance to fall armyworm (FAW; Spodoptera frugiperda) and fungal pathogens Colletotrichum graminicola and Cochliobolus heterostrophus. Metabolite profiling revealed that reduced resistance to insects and pathogens was associated with decreased production of PLVs and ketols, including the better studied α-ketol, 9,10-KODA (9-hydroxy-10-oxo-12(Z)-octadecadienoic acid). Exogenous PLV and 9,10-KODA treatments rescued the resistance of lox6 mutants to FAW and the pathogens. Surprisingly, the susceptible-to-herbivory lox6 mutants produced greater levels of wound-induced jasmonates, suggesting potential substrate competition between JA and PLV pathway branches and highlighting a strong role of PLVs in defense against insects. Similarly, likely due to substrate competition between GLV and PLV synthesis pathways, in response to C. graminicola infection, lox6 mutants accumulated elevated levels of GLVs, which promote susceptibility to this pathogen. Mutation of the GLV-producing ZmLOX10 in the lox6 mutant background reversed the susceptibility to C. graminicola, unveiling the contrasting roles of PLVs and GLVs in resistance to this pathogen. Overall, this study uncovered a potent signaling role of PLVs in defense against insect herbivory and fungal pathogens with distinct lifestyles., Competing Interests: Conflict of interest statement. The authors declare no conflict of interest., (© The Author(s) 2024. Published by Oxford University Press on behalf of American Society of Plant Biologists. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

X-rays irradiation has been demonstrated to effectively preserve the freshness of edible fungi and delay the loss of aroma during storage. In this study, shiitake mushrooms were irradiated with X-rays dose of 0.5 kGy and stored at 2 °C for 35 days. Non-irradiated mushrooms were recorded as control group. Results indicated that 0.5 kGy X-rays treatment preserved the flavor quality by exhibiting higher volatile substance content in shiitake mushrooms. X-rays treatment promoted the activities of lipoxygenase (LOX) and alcohol dehydrogenase (ADH) and oxidation of linoleic acid. In addition, the degradation of methionine and cysteine was facilitated by X-rays. Higher enzymes activities for γ-glutamyl transpeptidase (GGT) and cystine sulfoxide lyase (CS lyase) were found in 0.5 kGy X-rays irradiated mushrooms. These findings suggest that the retention of mushroom flavor by X-rays treatment is closely related to fatty acid metabolism, sulfur-containing amino acid metabolism, and lentinic acid metabolism., Competing Interests: Declaration of competing interest The authors confirm that they have no conflicts of interest with respect to the work described in this manuscript., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

A typical symptom of post-harvest pineapple (Ananas comosus L.) internal browning is influenced by multiple factors. However, the precise mechanism through which fruit size influences the occurrence of browning remains unclear. Therefore, this study aimed to investigate the impacts of fruit size in browning and its possible mechanisms in harvested pineapple fruit using physical and biochemical analysis and RNA-seq. Disease incidence was assessed in four groups of fruits (1, 1.5, 2, and 2.5 ± 0.2 kg), with the two most significant groups (1 and 2.5 ± 0.2 kg) selected for detailed analysis. The results showed that the large pineapple fruits had faster browning senescence and membrane lipid peroxidation, higher respiration intensity, more reactive oxygen species accumulation, and higher malondialdehyde content. Likewise, lower antioxidant capacity such as ascorbic acid, ascorbate peroxidase, catalase, and superoxide dismutase and higher polyphenol oxidase activity, peroxidase activity, phenylalanine ammonia-lyase activity, and lipoxygenase activity. It can be concluded that the large pineapple fruits were severely subjected to oxygen stress and membrane lipid peroxidation during storage. Gene ontology enrichment reveals that this relates mainly to oxidoreductase and glutathione metabolism. The large pineapple fruits accelerated AsA-GSH cycle pathway metabolism. Real-time quantitative PCR shows downregulated expression of AcAPX1, AcAPX6, AcAPX6 × 1, AcCAT2, AcSOD[Fe]2, and AcSODX2 in large pineapple fruits, whereas AcPPO and AcLOX upregulated expression. This study offers insights into fruit size and browning. Future molecular techniques may reduce fruit browning., (© 2025 Institute of Food Technologists.)

Background/Objectives: Abiotic stresses impose significant constraints on crop growth, development, and yield. However, the comprehensive characterization of the maize ( Zea mays ) lipoxygenase ( LOX ) gene family under stress conditions remains limited. LOXs play vital roles in plant stress responses by mediating lipid oxidation and signaling pathways. Methods: In this study, 13 ZmLOX genes were identified in maize and characterized to explore their functions under abiotic stresses. Results: Phylogenetics revealed that ZmLOX genes share evolutionary origins with LOX genes in Arabidopsis and rice. Promoter analysis identified cis -acting elements associated with growth, light response, hormone signaling, and stress response, indicating their diverse biological roles. Gene Ontology (GO) and KEGG enrichment analyses showed that ZmLOX genes are involved in jasmonic acid metabolism, lipid signaling, and photosynthetic processes, while protein-protein interaction (PPI) analysis positioned ZmLOX proteins as central hubs in stress-related regulatory networks. Differential expression and qRT-PCR analyses revealed stress-specific (including heat, drought, salt, and cold) expression patterns, with ZmLOX2 and ZmLOX13 showing key roles in drought and cold tolerance, respectively. Conclusions: These findings provide new insights into the regulatory functions of ZmLOX genes, offering potential targets for enhancing maize resilience to abiotic stresses and improving agricultural productivity.

The lipoxygenase (LOX) gene family is widely distributed in plants, and its activity is closely associated with seed viability and stress tolerance. In this study, we cloned the rice(Oryza sativa)lipoxygenase gene OsLOX1, a key participant in the 13-lipoxygenase metabolic pathway. Our primary focus was to investigate its role in mediating responses to drought stress and seed germination in rice. Histochemical staining and qPCR analysis indicated that the expression level of OsLOX1 was relatively high in leaves and early germinating seeds. Our findings revealed that mutant lines with CRISPR/Cas9-induced knockout of OsLOX1 exhibited reduced tolerance to drought stress compared with the wild-type. This was accompanied by elevated levels of H 2 O 2 and malondialdehyde, and a decrease in the expression levels of genes associated with antioxidant enzymes. Furthermore, knockout of OsLOX1 reduced the longevity of rice seeds increased H 2 O 2 and MDA levels, and decreased the activities of the antioxidant enzymes superoxide dismutase and catalase, compared with the wild-type. These findings demonstrated that OsLOX1 positively regulated rice seed vigor and drought stress., Competing Interests: Declarations. Conflict of interest: The authors declare no conflict of interest., (© 2024. The Author(s).)

Pelvic organ prolapse is a condition that significantly affects women's quality of life. The pathological mechanism of pelvic organ prolapse is not yet fully understood, and its pathogenesis is often caused by multiple factors, including the metabolic imbalance of the extracellular matrix. This study aims to investigate the role of miR-5195-3p, a microRNA, in the pathology of pelvic organ prolapse and its regulatory mechanism. Using various molecular biology techniques such as real-time reverse transcription Polymerase Chain Reaction (PCR), fluorescence in situ hybridization, immunohistochemistry, and Western blot, miR-5195-3p expression was examined in vaginal wall tissues obtained from pelvic organ prolapse patients. Results revealed an up-regulation of miR-5195-3p expression in these tissues, showing a negative correlation with the expression of extracellular matrix-related proteins. Further analysis using bioinformatics tools identified Lipoxygenase (LOX) as a potential target in pelvic organ prolapse. Dual luciferase reporter gene experiments confirmed LOX as a direct target of miR-5195-3p. Interestingly, regulating the expression of LOX also influenced the transforming growth factor β1 signaling pathway and had an impact on extracellular matrix metabolism. This finding suggests that miR-5195-3p controls extracellular matrix metabolism by targeting LOX and modulating the TGF-β1 signaling pathway. In conclusion, this study unveils the involvement of miR-5195-3p in the pathological mechanism of pelvic organ prolapse by regulating extracellular matrix metabolism through the LOX/TGF-β1 axis. These findings reveal new mechanisms in the pathogenesis of pelvic organ prolapse, providing a theoretical foundation and therapeutic targets for further research on pelvic organ prolapse treatment., (© The Author(s) 2024. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Polyphenolic plant compounds possess nutritional and pro-healthy potential, reducing the risk of auto-inflammatory and neoplastic diseases. However, their interference with the progression of thyroid gland dysfunctions has remained largely unaddressed. For this purpose, we combined the analyses of phenolic content and antioxidative activity with the thyroid peroxidase (TPO), lipoxygenase (LOX), xanthine oxidase (XO) and cyclooxygenase-2 (COX-2) activity assays, isobolographic approach and the estimation of thyroid cancer cells' proliferation and motility in vitro. Bioaccessible oatmeal (OM) and wheat flour (WF) compounds activated TPO while inhibiting LOX and XO's in vitro activity. OM extracts also inhibited COX-2 activity. Isobolographic and combination index studies revealed cooperation of compounds from OM and WF. However, the relatively strong inhibitory activity of bioaccessible OM compounds on LOX activity correlated with their mildly cytostatic and relatively distinct pro-invasive effects in the thyroid cancer model in vitro. Collectively, the application of OM and WF products for the prophylactics of inflammatory thyroid diseases should be considered with care, especially in the context of the oncological status of the patient., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

The aim of this study was to characterize and explore the food potential of sainfoin seeds, which are primarily used in their green plant form to feed livestock. A detailed assessment of the seed's proximate composition, enzyme activity levels, and antinutrient content was performed. The seeds were then subjected to infrared, hot air, and blanching treatments to examine how these processing methods affect enzyme activities and antinutrients. In addition, optimization of processing conditions was carried out using the response surface method and central composite design to ensure low antinutrient levels, minimal color change, and total enzyme inactivation. Sainfoin seeds have a crude protein content of 42 % in dry matter, making them a promising alternative plant-based protein source. They also contain 45 % dietary fiber, 9 % crude fat, and significant levels of potassium and magnesium. The unprocessed seeds exhibited 2.01 U/g lipase activity and 10.51 U/g lipoxygenase activity, which were inactivated with infrared treatment at 2000 W IR power in 75 s. In terms of antinutrients, sainfoin seeds contain 247 mg/100 g phytic acid, 2582 TIU/g trypsin inhibitor activity, 7.9 mg catechin equivalents of tannin /100 g, and 24.97 mg/100 g hydrocyanic acid. The optimal process conditions were 2000 W IR power and 105 s for IR treatment, 150 °C and 103 min for hot air treatment, and 81 °C and 16 min for blanching. The antinutrient levels and enzyme activities in unprocessed sainfoin seeds are comparable to those found in conventional legumes, although sainfoin seeds contain a higher amount of tannins. While infrared treatment was most effective for enzyme inactivation, the impact of the different processing methods varied across the parameters studied., 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.)

Eicosanoids and related compounds are pleiotropic lipid mediators, which play a role in cell differentiation and in the pathogenesis of various diseases. The biosynthesis of these lipids has extensively been studied in highly developed mammals including humans but little is known about the formation of these mediators in more ancient Prototheria. We searched the genomes of two extant prototherian species (platypus, short-beaked echidna) for genes encoding for lipoxygenase- (ALOX) and prostaglandin synthase-isoforms (PTGS) and detected intact single copy genes for ALOX5, ALOX12, ALOX12B, ALOXE3, PTGS1 and PTGS2. Moreover, we identified two copies of ALOX15B genes (ALOX15B-1 and ALOX15B-2) but in echidna the ALOX15B-2 gene was structurally corrupted. Interestingly, in the two genomes ALOX15 genes were lacking. For functional characterization we expressed the prototherian ALOX15B isoforms and compared important enzyme characteristics of the wildtype proteins and of relevant enzyme mutants with those of human and mouse ALOX15B. Here we observed that the prototherian ALOX15B isoforms exhibit the same reaction specificity as their human ortholog. Mutagenesis of the Triad determinants did not alter the reaction specificity of the prototherian enzymes but modification of the Jisaka determinants murinized the catalytic properties. These data indicate that Prototheria exhibit an active eicosanoid metabolism. They express functional ALOX15B orthologs but lack ALOX15 genes. These observations and the previous findings that ALOX15 orthologs rarely occur in non-mammalian vertebrates such as fish and birds suggest that ALOX15 orthologs were introduced during Prototheria-Metatheria transition via an ALOX15B gene duplication and subsequent divergent enzyme evolution., Competing Interests: Declaration of competing interest The authors declare that they do not have any conflicts of interest with the content of this publication., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Lipoxygenases (LOXs) catalyze the regioselective dioxygenation of polyunsaturated fatty acids (PUFAs), generating fatty acid hydroperoxides (FAHPs) with diverse industrial applications. Bacterial LOXs have garnered significant attention in recent years due to their broad activity towards PUFAs, yet knowledge about the structural factors influencing their substrate preferences remains limited. Here, we characterized a bacterial LOX from Burkholderia thailandensis (Bt-LOX), and identified key residues affecting its substrate preference and regioselectivity through site-directed mutagenesis. Bt-LOX preferred ω-6 PUFAs and exhibited regioselectivity at the ω-5 position. Mutations targeting the substrate binding pocket and the oxygen access channel led to the production of three active variants with distinct catalytic properties. The A431G variant bifurcated dioxygenation between the ω-5 and ω-9 positions, while F446V showed reduced regioselectivity with longer PUFAs. Interestingly, L445A displayed altered substrate specificity, favoring ω-3 over ω-6 PUFAs. Furthermore, L445A shifted the regioselectivity of dioxygenation to the ω-2 position in ω-3 PUFAs, and, for some substrates, facilitated dioxygenation closer to the carboxylic acid terminus, suggesting an altered substrate orientation. Among these variants, L445A represents a significant milestone in LOX research, as these alterations in substrate specificity, dioxygenation regioselectivity, and substrate orientation were achieved by a single mutation only. These findings illuminate key residues governing substrate preference and regioselectivity in Bt-LOX, offering opportunities for synthesizing diverse FAHPs and highlighting the potential of bacterial LOXs as biocatalysts with widespread applications., Competing Interests: Declaration of Competing Interest None., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Osmotic stress, caused by the lack of water or by high salinity, is a common problem in plant roots. Osmotic stress can be reproducibly simulated with the application of solutions of the high-molecular-weight and impermeable polyethylene glycol. The accumulation of different reactive oxygen species, such as singlet oxygen, superoxide, and hydrogen peroxide, accompany this stress. Among them, singlet oxygen, produced as a byproduct of lipoxygenase activity, has been associated with limiting root growth. To better understand the source and effect of singlet oxygen, we followed its production at the cellular level in Arabidopsis (Arabidopsis thaliana). Osmotic stress initiated profound changes in plastid and vacuole structure. Confocal and electron microscopy showed that the plastids were a source of singlet oxygen accompanied by the appearance of multiple, small extraplastidic bodies that were also an intense source of singlet oxygen. A marker protein, CRUMPLED LEAF, indicated that these small bodies originated from the plastid outer membrane. Remarkably, LINOLEATE 9S-LIPOXYGENASE 5 (LOX5) was shown to change its distribution from uniformly cytoplasmic to a more clumped distribution together with plastids and the small bodies. In addition, oxylipin products of Type 9 lipoxygenase increased, while products of Type 13 lipoxygenases decreased. Inhibition of lipoxygenase by the salicylhydroxamic acid inhibitor or in downregulated lipoxygenase lines prevented cells from initiating the cellular responses, leading to cell death. In contrast, singlet oxygen scavenging halted terminal cell death. These findings underscore the reversible nature of osmotic stress-induced changes, emphasizing the pivotal roles of lipoxygenases and singlet oxygen in root stress physiology., Competing Interests: Conflict of interest statement. None declared., (© The Author(s) 2024. Published by Oxford University Press on behalf of American Society of Plant Biologists. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Key Message: Lipoxygenase activity and localization vary throughout the development of Larix kaempferi ovules, with the highest enzyme activity observed in ovules at the cellular stage and the most intense immunogold reaction noted at the mature archegonium stage of gametophyte development. Lipoxygenases are a family of oxidoreductases with a significant role in biological systems, widespread in living organisms e.g. mammals, fish, corals, plants, mosses, algae, fungi, yeasts, and bacteria. Lipoxygenase activity in plants leads to the formation of phytooxylipins, i.e. signaling molecules, which play a crucial role in many significant physiological processes such as male and female gametophyte maturation, germination and seedling growth, pathogen resistance, abiotic stress response, fruit ripening, and senescence. The activity and localization of lipoxygenase change during plant growth and development. The localization of lipoxygenase in a developing ovule of Larix kaempferi was analyzed using the immunogold labeling method, and the activity was determined spectrophotometrically with linolenic acid as a substrate. Among the investigated stages, the immunogold reaction was the most intense at the mature archegonium stage in the ovule. Lipoxygenase was found in all parts of the L. kaempferi ovule. The largest number of immunogold particles was detected in the integument cells of all the analyzed stages of ovule development. Only one isoform of lipoxygenase with an optimum at pH 8 was active in the ovules during female gametophyte maturation. The highest enzyme activity was determined at the cellular stage, whereas the mature archegonium stage was characterized by its lowest level, which means that LOX activity in developing ovules of the Japanese larch is not correlated with the number of antibody-labeled molecules of the enzyme., (© 2024. The Author(s).)

In this paper, we attempted to develop a novel class of compounds against lipoxygenase, a key enzyme in the biosynthesis of leukotrienes implicated in a series of inflammatory diseases. Given the absence of appropriate human 5-lipoxygenase crystallographic data, solved soybean lipoxygenase-1 and -3 structures were used as a template to generate an accurate pharmacophore model which was further used for virtual screening purposes. Eight compounds (1-8) have been derived from the in-house library consisting of N-substituted pyrroles conjugated with 5- or 6-indazole moieties through a carboxamide linker. This study led to the discovery of hit molecule 8 bearing a naphthyl group with the IC 50 value of 22 μM according to soybean lipoxygenase in vitro assay. Isosteric replacement of naphthyl ring with quinoline moieties and reduction of carbonyl carboxamide group resulted in compounds 9-12 and 13, respectively. Compound 12 demonstrated the most promising enzyme inhibition. In addition, compounds 8 and 12 were found to reduce the carrageenan-induced paw edema in vivo by 52.6 and 49.8%, respectively. In view of the encouraging outcomes concerning their notable in vitro and in vivo anti-inflammatory activities, compounds 8 and 12 could be further optimized for the discovery of novel 5-lipoxygenase inhibitors in future., Competing Interests: Declarations. Conflicts of interest: The authors declare no conflict of interest. Ethical approval: The animal study protocol was approved by the Ethics Committee of the Prefecture of Central Macedonia (protocol code 270079/2500) for studies involving animals. Human and animal rights: No humans were used for studies that are the basis of this research. All the animal experiments complied with the ARRIVE guidelines and were in accordance with the ethical standards of the European Directive 2010/63/EU on the protection of animals used for scientific purposes. Informed consent: Not applicable., (© 2023. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

The lipoxygenase (LOX) activity of major tropical fish used for surimi production, including threadfin bream (TB), lizardfish (LZ), and goatfish (GF), was measured in the gills, skin, and muscle. The highest LOX activity was observed in the LZ samples (p < 0.05), with the gills exhibiting the greatest activity at 376.56 U/mg (p < 0.05). The highest peroxide value was detected in the TB samples, particularly in the gills (p < 0.05). Docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) were the main polyunsaturated fatty acids in all the tissues and surimi. The total lipid and DHA contents of washed mince reduced considerably after the screw press process. Although LOX activity decreased during surimi production, a residual activity of 21.33 U/g was observed in the finished surimi. LOX was partially purified and characterized from LZ gills. The purification was conducted using two successive chromatographic steps, Sephacryl S-200 and diethylaminoethyl (DEAE)-sepharose, resulting in a 3.52% yield and a 22.43-fold increase in purity. The optimum activity was found at 25°C and pH 7.5, with pH stability between 6.0 and 8.5. The relatively high thermal stability at 4°C-10°C suggested that LOX might contribute to fish lipid oxidation during cold storage. The enzyme was thermally inactivated at 60°C. The preferred substrate was EPA. LOX from the LZ gills was inhibited by 1 mM ethylenediaminetetraacetic acid and activated by 1 mM Fe 2+ , Na + , and Ca 2+ . PRACTICAL APPLICATION: Elucidating lipoxygenase activity and lipid oxidation in various tropical fish tissues, as well as understanding the characteristics of LOX, can help take appropriate postharvest actions to afford high-quality surimi., (© 2024 Institute of Food Technologists.)

Human arachidonate 15-lipoxygenase type B is a lipoxygenase that catalyzes the peroxidation of arachidonic acid at carbon-15. The corresponding murine ortholog however has 8-lipoxygenase activity. Both enzymes oxygenate polyunsaturated fatty acids in S-chirality with singular reaction specificity, although they generate a different product pattern. Furthermore, while both enzymes utilize both esterified fatty acids and fatty acid hydro(pero)xides as substrates, they differ with respect to the orientation of the fatty acid in their substrate-binding pocket. While ALOX15B accepts the fatty acid "tail-first," Alox8 oxygenates the free fatty acid with its "head-first." These differences in substrate orientation and thus in regio- and stereospecificity are thought to be determined by distinct amino acid residues. Towards their biological function, both enzymes share a commonality in regulating cholesterol homeostasis in macrophages, and Alox8 knockdown is associated with reduced atherosclerosis in mice. Additional roles have been linked to lung inflammation along with tumor suppressor activity. This review focuses on the current knowledge of the enzymatic activity of human ALOX15B and murine Alox8, along with their association with diseases., Competing Interests: Declarations. Ethical approval: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Prostate cancer (PCa) has become a common disease among middle-aged and elderly men. The lipoxygenase (LOX) pathway plays a crucial role in the occurrence, development, invasion and metastasis of PCa and is therefore considered a new target for the prevention and treatment of PCa. 5-LOX and 12-LOX have a promoting effect on the occurrence, development, invasion and metastasis of PCa. 15-LOX-2 has an inhibitory effect on PCa. LOX inhibitors can effectively inhibit the metabolic activity of LOX. The research aims to review the mechanism of action and inhibitors of LOX in PCa, in order to provide relevant references for the prevention and treatment of PCa.

Asthma is a chronic allergic respiratory disease lacking effective therapies. The present study investigated the anti‑asthmatic properties of lipid mediators using an ovalbumin (OVA)‑induced allergic asthma model. Lipid mediators (LM; 17S‑monohydroxy docosahexaenoic acid, resolvin D5 and protectin DX at a ratio of 3:47:50) were derived from docosahexaenoic acid through soybean lipoxygenase. LM treatment significantly alleviated major features of allergic asthma, including inflammatory cell infiltration, with a particular reduction in eosinophils in bronchoalveolar lavage fluid, downregulation of Th2 cytokine expression, attenuation of airway remodeling, and oxidative stress, thereby closely resembling the normal condition. Additionally, a significant increase in the serum levels of interleukin‑6 [167.12±6.25 pg/ml; P<0.0001 vs. negative control (NC) group], tumor necrosis factor‑α (109.17±7.17 pg/ml; P<0.0001 vs. NC group) and IgE (90.24±5.98 ng/ml; P<0.0001 vs. NC group) was observed following OVA challenge; however, oral administration of LM resulted in a notable reduction in these levels to 99.45±6.12 pg/ml (P<0.001 vs. OVA group), 62.51±4.03 pg/ml (P<0.001 vs. OVA group) and 56.50±2.70 ng/ml (P<0.001 vs. OVA group), respectively. Furthermore, the heightened expression of Th2‑related cytokines induced by OVA was observed to be restored closely to normal conditions following LM treatment, as demonstrated for both gene and protein expression levels. Histological analysis demonstrated that LM mitigated inflammatory cell infiltration while reducing mucus secretion. Additionally, LM effectively ameliorated oxidative stress in OVA‑induced asthma, with a significant increase in the activity of superoxide dismutase (~185% vs. OVA group; P<0.001), elevated levels of glutathione (~74% higher than the OVA group; P<0.001) and reduced content of malondialdehyde (~40% lower than the OVA group; P<0.001) in lung tissues. Collectively, these findings suggested that LM effectively protected lung tissues from inflammation and oxidative stress, thereby representing a promising therapeutic option for the treatment of allergic asthma.

The regulation of reaction rate differentiation, catalytic precursor differentiation, and end-product differentiation during enzyme-mediated reactions within complex lipid systems is a key area of research in flavor regulation. A multilayer lipid oxidation model, utilizing Plaice bone oil (PBO), lipase, and lipoxygenase, was employed to investigate oxidation differences between various lipids and corresponding flavor formation patterns. Lipase treatment resulted in higher levels of non‑oxygenated volatile compounds and saturated aldehydes, whereas lipoxygenase treatment increased oxygenated compounds, particularly (E)-2-hexenal, 1-penten-3-one, and 2-pentylfuran. The hydrolysis of triglycerides by lipase significantly raised the concentration of monounsaturated fatty acids. Lipoxygenase catalyzed the formation of position-specific oxidation products at the ω-6 carbon position of PUFAs, such as 15-hydroperoxy-EPA and 17-hydroperoxy-DHA. These enzymatic treatments altered the lipid profile, leading to distinct flavor formation patterns. This study provides valuable insights into the mechanisms underlying enzyme-mediated volatile substance variations in food., 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.)

Oxylipins are signaling lipids derived from the oxidation of polyunsaturated fatty acids (PUFAs). In lipidomic studies, human plasma may be subjected to various storage conditions and freeze-thaw cycles, which may impact the analysis of these compounds. In this study, we used liquid chromatography coupled with mass spectrometry (LC-MS) to examine the influence of up to five freeze-thaw cycles (FTCs) on free and total (mostly esterified) oxylipins in human plasma and the influence of temperature and storage duration (4 °C for up to 120 h and -20 °C and -80 °C for 1-98 days) in the presence or absence of butylated hydroxytoluene (BHT) on extracted oxylipins stored in LC-MS amber vials. In fresh plasma subjected to several FTCs, approximately 48 % of the detected free oxylipins were significantly altered by the third cycle, with increases in cytochrome P450 (CYP450) and lipoxygenase (LOX)-derived compounds and reductions in trihydroxylated oxylipins. In contrast, multiple FTCs did not significantly alter esterified oxylipins. At 4 °C, the extracted oxylipins did not change significantly for up to 120 h (5 days). Oxylipin levels remained stable for 98 days at -80 °C but decreased by 98 days at -20 °C. The antioxidant activity of butylated hydroxytoluene (BHT) did not influence oxylipin stability at 4 °C for 120 h or at -80 °C for 98 days, but it reduced oxylipin degradation at -20 °C at 98 days. Conversely, prostaglandin F 2α (PGF 2α ) exhibited substantial increases at -20 °C and -80 °C, independent of BHT. This study demonstrates that (i) unlike free oxylipins, the esterified oxylipin pool remains stable following repeated FTCs, (ii) extracted oxylipins are stable at 4 °C for up to 120 h and at -80 °C for up to 98 days, but not at -20 °C for 98 days, and (iii) BHT may minimize oxylipin degradation of sample extracts stored at -20 °C. This study provides a framework for measuring oxylipins under various freeze-thaw and storage conditions., 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: Lipoxygenases (LOXs) are key enzymes in the unsaturated fatty acid oxidation reaction pathway and play an important regulatory role in the synthesis of fruit aroma volatiles., Methods: LOX gene family members were identified in the whole genome database of bitter gourd and analyzed bioinformatically. An RT-qPCR was used to analyze the expression differences in different tissues. Monoterpenes were determined by gas chromatography-mass spectrometry (GC-MS) technique., Results: A total of 12 LOX gene family members were identified in the genome. The expression of LOX genes varied significantly among the tissues of roots, stems, leaves, flowers, fruits, seeds and tendrils. A total of 29 monoterpenes were detected in the fruits of five different fruit colors of bitter gourd, mainly containing six types of alcohols, aldehydes, terpenes, ketones, esters and alkynes, with the highest relative content of alcohols., Conclusions: The present study provides a reference for further elucidation of the biological functions of the LOX gene in the synthesis pathway of aroma volatiles in bitter gourd.

Iron-dependent lipoxygenases (LOXs) are involved in the synthesis of oxylipins from polyunsaturated fatty acids. However, they are usually difficult to overexpress in functional form in microbial cell factories. Moreover, 9-LOXs, generating 9-hydroperoxy fatty acids from C18 polyunsaturated fatty acids, have rarely been found from microbial sources. Here, we discovered a novel 9 S -LOX in the marine myxobacterium Enhygromyxa salina (Es-9 S -LOX). The recombinant enzyme produced in Escherichia coli exhibited remarkable activity in the dioxygenation of linoleic acid (LA, 1 ), α-linolenic acid, γ-linolenic acid, and arachidonic acid specifically at the C9 position to yield the product with ( S )-configuration at catalytic efficiency of 3.94, 1.42, 1.38, and 0.69 μM -1 ·s -1 , respectively. The elucidated X-ray crystal structure of Es-9 S -LOX reveals a long and narrow hydrophobic pocket that allows the substrate to be near the metal ion and the oxygen tunnel. The enzyme was successfully used in a chemoenzymatic reaction to generate a hydroxy fatty acid from LA. Our study thus contributes to the valorization of renewable polyunsaturated fatty acids into a variety of fatty acid derivatives, including hydroxy fatty acids.

Recent research has highlighted the pivotal role of lipoxygenases in modulating ferroptosis and immune responses by catalyzing the generation of lipid peroxides. However, the limitations associated with protein enzymes, such as poor stability, low bioavailability, and high production costs, have motivated researchers to explore biomimetic materials with lipoxygenase-like activity. Here, we report the discovery of lipoxygenase-like two-dimensional (2D) MoS 2 nanosheets capable of catalyzing lipid peroxidation and inducing ferroptosis. The resulting catalytic products were successfully identified using mass spectrometry and a luminescent substrate. Unlike native lipoxygenases, MoS 2 nanosheets exhibited exceptional catalytic activity at extreme pH, high temperature, high ionic strength, and organic solvent conditions. Structure-activity relationship analysis indicates that sulfur atomic vacancy sites on MoS 2 nanosheets are responsible for their catalytic activity. Furthermore, the lipoxygenase-like activity of MoS 2 nanosheets was demonstrated within mammalian cells and animal tissues, inducing distinctive ferroptotic cell death. In summary, this research introduces an alternative to lipoxygenase to regulate lipid peroxidation in cells, offering a promising avenue for ferroptosis induction.

We propose a co-immobilized chemo-enzyme cascade system to mitigate random intermediate diffusion from the mixture of individual immobilized catalysts and achieve a one-pot reaction of multi-enzyme and reductant. Catalyzed by lipase and lipoxygenase, unsaturated lipid hydroperoxides (HPOs) were synthesized. 13(S)-hydroperoxy-9Z, 11E-octadecadienoic acid (13-HPODE), one compound of HPOs, was subsequently reduced to 13(S)-hydroxy-9Z, 11E-octadecadienoic acid (13-HODE) by cysteine. Upon the optimized conditions, 75.28 mg of 13-HPODE and 4.01 mg of 13-HODE were produced from per milliliter of oil. The co-immobilized catalysts exhibited improved yield compared to the mixture of individually immobilized catalysts. Moreover, it demonstrated satisfactory durability and recyclability, maintaining a relative HPOs yield of 78.5% after 5 cycles. This work has achieved the co-immobilization of lipase, lipoxygenase and the reductant cysteine for the first time, successfully applying it to the conversion of soybean oil into 13-HODE. It offers a technological platform for transforming various oils into high-value products., 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 © 2023. Published by Elsevier Ltd.)

Lipoxygenases catalyze the peroxidation of poly-unsaturated fatty acid chains either free or esterified in membrane lipids. Vitis vinifera LoxA is transcriptionally induced at ripening onset and localizes at the inner chloroplast membrane where it is responsible for galactolipid regiospecific mono- and di-peroxidation. Here we present a kinetic and structural characterization of LoxA. Our X-ray structures reveal a constitutive dimer with detergent induced conformational changes affecting substrate binding and catalysis. In a closed conformation, a LID domain prevents substrate access to the catalytic site by steric hindrance. Detergent addition above the CMC destabilizes the LID and opens the dimer with both catalytic sites accessible from the same surface framed by the PLAT domains. As a consequence, detergent molecules occupy allosteric sites in the PLAT/catalytic domain interface. These structural changes are mirrored by increased enzymatic activity and positive cooperativity when the substrate is provided in micelles. The ability to interact with micelles is lost upon dimer destabilization by site-directed mutagenesis as assessed by tryptophan fluorescence. Our data allow to propose a model for protein activation at the membrane, classifying LoxA as an interfacial enzyme acting on fatty acid chains directly from the membrane similar to mammalian 15-LOX and 5-LOX., 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 aim of the presented study was to evaluate the release of the enzymatically initiated production of hexanal from double emulsion electrospun bio-active membranes at a temperature of fruit storage. Among different formulations of water-in-oil (W1/O) primary emulsions, the emulsion composed of 12% w/v Tween20 and 0.1 M NaCl in water (W1) and 6% of poly(glycerol) poly(ricinoleate) dissolved in sunflower oil (O) using W1/O ratio of 80/20 (w/w) (Tween20-NaCl/6% PGPR) was selected, for further incorporation of enzymes, based on the lowest average droplet size (391.0 ± 15.6 nm), low polydispersity index (0.255 ± 0.07), and good gravitational stability also after 14 days. Both enzymes, lipase and lipoxygenase are needed to produce hexanal (up to 58 mg/L). Additionally, double emulsions were prepared with sufficient conductivity and viscosity using different W1/O to W2 ratios for electrospinning. From the selected electrospun membrane, up to 4.5 mg/L of hexanal was released even after 92 days., 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.)

Senescence represents a developmentally orchestrated and precisely regulated cascade of events, culminating in the abscission of plant organs and ultimately leading to the demise of the plant or its constituent parts. In this study, we observed that senescence in cut Lilium tigrinum flowers is induced by elevated ABA levels and the hyperactivation of lipoxygenase (LOX) activity. This cascade increased ROS concentrations, heightened oxidative damage, and disrupted cellular redox equilibrium. This was evidenced by elevated lipid peroxidation, attenuated antioxidant machinery, and reduced membrane stability index (MSI). Despite its known role in delaying flower senescence, the specific biochemical and molecular mechanisms by which nitric oxide (NO) regulates senescence in cut L. tigrinum flowers are not fully elucidated. Specifically, the interactions between NO signaling and ABA metabolism, the regulation of protease activity, and the influence of NO-mediated ROS scavenging, senescence-associated gene expression requires further exploration. Exogenous application of sodium nitroprusside (SNP), a source of NO, mitigated senescence in L. tigrinum cut flowers by upregulating the activity of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and reducing the LOX activity, an indicator of lipid peroxidation. SNP treatment also downregulated the relative expression of senescence-associated gene (SAG12 ),lipoxygenase 1 (LOX1 ), and abscisic aldehyde oxidase 3 (AAO3 ). NO also upregulated defender against apoptotic death 1 (DAD1 ) expression correlated with minimized protease activity and reduced α-amino acid content in SNP-treated tepals. This regulation was accompanied by increased contents of sugars, proteins and phenols and reduced abscisic acid content, which collectively delayed the senesecence and enhanced the longevity of L. tigrinum cut flowers. This study demonstrates that exogenous SNP application can effectively mitigate senescence in cut L. tigrinum flowers by modulating antioxidant enzyme activities, reducing oxidative stress, and regulating the expression of key senescence-associated genes. This study unravels the complex molecular networks involved in NO-mediated senescence delay, which may lead to the development of innovative approaches for improving flower longevity.

Longan fruit deteriorates rapidly after harvest, which limits its storability. This study aimed to investigate the effect of tert-butylhydroquinone (TBHQ) on quality maintenance, membrane lipid metabolism, and energy status of longan fruit during 25 °C storage. Compared with control fruit, TBHQ treatment maintained better marketable fruit rate and suppressed activities of phospholipase D (PLD), lipase, and lipoxygenase (LOX), and downregulated expressions of DlPLD, DlLOX, and Dllipase. TBHQ also increased the ratio of unsaturated fatty acids to saturated fatty acids (U/S) and the index of unsaturated fatty acids (IUFA). In addition, higher levels of ATP, ADP, energy charge, NADP + / NADPH as well as higher activities of H + -ATPase, Ca 2+ -ATPase and NADK were also observed in TBHQ-treated fruit. These results suggested that TBHQ may maintain postharvest quality of longan fruit by regulating membrane lipid and energy metabolisms., 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.)

9-Carbon aldehydes such as (2E)-nonenal, (3Z)-nonenal, and (2E,6Z)-nonadienal are important melon and cucumber fragrance compounds. Currently, these molecules are produced either synthetically, which faces consumer aversion, or through biotransformation using plant-extracted enzymes, which is costly and inefficient. In this study, we constructed a Saccharomyces cerevisiae platform for the whole cell biotransformation of polyunsaturated fatty acids (PUFAs) to 9-carbon aldehydes. Heterologous expression of lipoxygenase (LOX) from Nicotiana benthamiana and hydroperoxide lyase (HPL) from Cucumis melo (melon) in S. cerevisiae enabled the production of (2E)-nonenal from readily available polyunsaturated fatty acid substrates. A 5.5-fold increase in (2E)-nonenal titer was then achieved utilizing genetic and reaction condition enhancement strategies. The highest titer of (2E)-nonenal was more than 0.11 mM, with about 9% yield. This platform can potentially be used to produce a variety of other aldehyde products by customizing with LOX and HPL enzymes of different regio-selectivities. KEY POINTS: • Establishment of a S. cerevisiae whole-cell biotransformation platform for cost-efficient, high-yield conversion of PUFAs into high value 9-carbon aldehyde compounds • 5.5-Fold improvement of (2E)-nonenal titer to > 0.11 mM achieved by enhancing reaction conditions and gene expression levels of LOX and HPL., (© 2024. The Author(s).)

The study investigated the impact of UV-C irradiation on peach fruit quality during postharvest storage, with a focus on aroma changes and the mechanisms involving lipoxygenase metabolism. Results showed that UV-C irradiation at a dosage of 1.5 kJ/m 2 was found to preserve the quality attributes of peach fruit during ambient storage, as evidenced by high flesh firmness, inhibition of weight loss and respiration rate, as well as high values of L* and ascorbic acid. Meanwhile, UV-C irradiation led to an increase in the contents of aroma-related volatiles, particularly esters and lactones, compared to non-irradiated fruit. Our results suggested that the enhanced emission of aroma-related volatiles in UV-C irradiated peach fruit was linked to elevated levels of unsaturated fatty acids. Besides, UV-C induced the expressions and activities of enzymes in the lipoxygenase pathway, thus promoting the synthesis of esters and lactones, which contribute to the enhanced aroma in peach fruit., 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.)

Lipoxygenases (LOXs) from lower organisms have substrate flexibility and function versatility in fatty acid oxidation, but it is not clear how these LOXs acquired the ability to execute multiple functions within only one catalytic domain. This work studied a multifunctional LOX from red alga Pyropia haitanensis (PhLOX) which combined hydroperoxidelyase (HPL) and allene oxide synthase (AOS) activity in its active pocket. Molecular docking and site-directed mutagenesis revealed that Phe642 and Phe826 jointly regulated the double peroxidation of fatty acid, Gln777 and Asn575 were essential to the AOS function, and the HPL activity was improved when Asn575, Gln777, or Phe826 was replaced by leucine. Phylogenetic analysis indicated that Asn575 and Phe826 were unique amino acid sites in the separated clades clustered with PhLOX, whereas Phe642 and Gln777 were conserved in plant or animal LOXs. The N-terminal START/RHO&#95;alpha&#95;C/PITP/Bet&#95;v1/CoxG/CalC (SRPBCC) domain of PhLOX was another key variable, as the absence of this domain disrupted the versatility of PhLOX. Moreover, the functions of two homologous LOXs from marine bacterium Shewanella violacea and red alga Chondrus crispus were examined. The HPL activity of PhLOX appeared to be inherited from a common ancestor, and the AOS function was likely acquired through mutations in some key residues in the active pocket. Taken together, our results suggested that some LOXs from red algae attained their versatility by amalgamating functional domains of ancestral origin and unique amino acid mutations.

Inflammation is the preliminary response given to any possible harmful stimuli including infections, injury or stress by immune system where neutrophils and macrophages gets activated and produces mediators, such as nitric oxide and cytokines that serves as biomarkers of inflammation. Lipoxygenases are enzymes that peroxidises lipids and are involved in the pathogenesis of several diseases including inflammatory diseases. These are oxidative enzymes comprising a non-heme iron atom in active site and are convoluted in inflammatory reactions. Fucoidan is sulphated polysaccharide that has numerous pharmacological implications. Implications of fucoidan on inflammatory diseases are still an objective of rigorous research. Therefore, this study focusses on investigating lipoxygenase inhibitory activities of fucoidan. The mechanism of lipoxygenase inhibitory activities of fucoidan was studied via molecular docking and molecular dynamics simulations. The docking score produced by the binding of the fucoidan to the lipoxygenase was - 6.69 kcal/mol whereas, the docking score in case of Aspirin and Zileuton were -5.8 kcal/mol and -7.0 kcal/mol and it was found that fucoidan makes hydrogen bonds with lipoxygenase protein through polar amino acid glutamine at GLN 514. The results obtained from molecular dynamics simulations proposed the development of a stable complex between fucoidan and lipoxygenase due to the establishment of favourable interactions with amino acid residues and indicated efficient results when compared with Aspirin and Zileuton. This study suggested that fucoidan had anti-inflammatory potentials and thus can be used as a promising drug candidate against inflammation.Communicated by Ramaswamy H. Sarma.

The structural and digestive properties of indica rice starch-fatty acid complexes and the effects of lipoxygenase on the structural and digestive properties of the complexes were examined in this study. The complexes were characterized by scanning electron microscopy, X-ray diffraction, Fourier transform-infrared spectroscopy and Raman spectroscopy. The results showed that indica rice starch had the highest molecular chain order and the highest crystallinity, and the crystallization disappeared after gelatinization, and the formation of indica rice starch-fatty acid complexes promoted the transformation of starch crystal structure from A-type to V-type. Lipoxygenase reduced the regularity of starch molecular crystal structure in the complexes, while enzyme protein improved the order of starch molecular structure in the complexes. The regularity of starch crystal structure in the complexes could improve with the increase of composite temperature and the increase of fatty acid unsaturation. In vitro digestibility and in vitro digestion kinetics showed that the formation of indica rice starch-fatty acid complexes reduced the digestibility of indica rice starch to a certain extent. The RDS content of indica rice starch was 66.42 ± 0.39 %, and lipoxygenase reduced the reduction of rapidly digested starch content during complexes digestion, while enzyme protein increased the content of resistant starch., 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.)

Manganese hydroxido (Mn-OH) complexes supported by a tripodal N,N',N″-[nitrilotris(ethane-2,1-diyl)]tris(P,P-diphenylphosphinic amido) ([poat] 3- ) ligand have been synthesized and characterized by spectroscopic techniques including UV-vis and electron paramagnetic resonance (EPR) spectroscopies. X-ray diffraction (XRD) methods were used to confirm the solid-state molecular structures of {Na 2 [Mn II poat(OH)]} 2 and {Na[Mn III poat(OH)]} 2 as clusters that are linked by the electrostatic interactions between the sodium counterions and the oxygen atom of the ligated hydroxido unit and the phosphinic (P=O) amide groups of [poat] 3- . Both clusters feature two independent monoanionic fragments in which each contains a trigonal bipyramidal Mn center that is comprised of three equatorial deprotonated amide nitrogen atoms, an apical tertiary amine, and an axial hydroxido ligand. XRD analyses of {Na[Mn III poat(OH)]} 2 also showed an intramolecular hydrogen bonding interaction between the Mn III -OH unit and P=O group of [poat] 3- . Crystalline {Na[Mn III poat(OH)]} 2 remains as clusters with Na + ---O interactions in solution and is unreactive toward external substrates. However, conductivity studies indicated that [Mn III poat(OH)] - generated in situ is monomeric and reactivity studies found that it is capable of cleaving C-H bonds, illustrating the importance of solution-phase speciation and its direct effect on chemical reactivity. Synopsis: Manganese-hydroxido complexes were synthesized to study the influence of H-bonds in the secondary coordination sphere and their effects on the oxidative cleavage of substrates containing C-H bonds., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024. Published by Elsevier Inc.)

Background: 1.5 million new HIV infections occurred in 2021, suggesting new prevention methods are needed. Inflammation increases the risk for HIV acquisition by attracting HIV target cells to the female genital tract (FGT). In a pilot study, acetylsalicylic acid (ASA/Aspirin) decreased the proportion of FGT HIV target cells by 35 %. However, the mechanism remains unknown., Methods: Women from Nairobi, Kenya took low-dose ASA (81 mg) daily for 6-weeks. Free oxylipins in the plasma were quantified by high-performance liquid chromatography-tandem mass spectroscopy., Results: Oxylipins from 9 fatty acid substrates were detected, with more than one analyte from 4 substrates reduced post-ASA. Summary analysis found ASA downregulated cyclooxygenase and lipoxygenase but not cytochrome P450 activity with a lower n-6/n-3 oxylipin profile, reflecting reduced inflammation post-ASA., Conclusions: Inflammation is associated with increased lipoxygenase activity and HIV risk. Our data suggests ASA reduces inflammation through downregulation of oxylipins. Understanding how ASA reduces inflammation may lead to novel HIV prevention approaches., Competing Interests: Declaration of Competing Interest The authors have no conflicts of interest to declare., (Copyright © 2024. Published by Elsevier Inc.)

The natural aroma compound (+)-nootkatone was obtained in selective conversions of up to 74 mol% from inexpensive (+)-valencene substrate by using a comparatively greener biocatalytic process developed based on modifications of the previously published Firmenich method. Buffer identity and concentration, pH, temperature and downstream work-up procedures were optimized to produce a crude product in which >90 % of (+)-valencene had been converted, with high chemoselectivity observed for (+)-nootkatone production. Interestingly, the biotransformation was carried out efficiently at temperatures as low as 21 ºC. Surprisingly, the best results were obtained when an acidic pH in the range of 3-6 was applied, as compared to the previously published procedure in which it appeared to be necessary to buffer the pH optimally and fixed throughout at 8.5. Furthermore, there was no need to maintain a pure oxygen atmosphere to achieve good (+)-nootkatone yields. Instead, air bubbled continuously at a low rate through the reaction mixture via a submerged glass capillary was sufficient to enable the desired lipoxygenase-catalyzed oxidation reactions to occur efficiently. No valencene epoxide side-products were detected in the organic product extract by a standard GCMS protocol. Only traces of the anticipated corresponding α- and β-nootkatol intermediates were routinely observed., 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.)

Nine-carbon aldehydes and their relative alcohols (C9 aromas) are the main aroma compounds of cucumber (Cucumis sativus L.) fruits and provide a unique cucumber-like note. However, the key regulators of C9 aroma accumulation in cucumber fruit are poorly characterized. Based on C9 aroma dynamic analysis and transcriptome analysis during fruit development of two different cucumber inbred lines, Q16 and Q24, Lipoxygenase09 (CsLOX09) was identified as a candidate gene for C9 aroma accumulation. Additionally, Q24 with higher CsLOX09 expression accumulated more C9 aromas than Q16. To verify the function of CsLOX09, Cslox09 homozygote knockout lines were created. C9 aroma content decreased by 80.79% to 99.16% in these mutants compared to the wild type. To further explore the reasons for the difference in CsLOX09 expression between Q16 and Q24 fruits, a co-expression network was constructed by integrating the C9 aroma-associated metabolism and transcriptomic data. Eighteen candidate transcription factors were highly correlated with the expression of CsLOX09. DNA binding with One Finger 1.8 (CsDof1.8) was confirmed to bind directly to the A/TAAAG motif of the CsLOX09 promoter through dual-luciferase, yeast one-hybrid, chromatin immunoprecipitation-qPCR and electrophoretic mobility shift assays. Furthermore, C9 aroma content and CsLOX09 expression were significantly increased in the CsDof1.8 overexpression lines. Overall, these data elucidate the metabolic regulation of C9 aromas in cucumber and provide a foundation for facilitating the regulation of flavor in cucumber breeding., Competing Interests: Conflict of interest statement. The authors declare that they have no conflicts of interest., (© The Author(s) 2024. Published by Oxford University Press on behalf of American Society of Plant Biologists. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Immune-priming occurs in insects after a prior pathogen exposure. However, its underlying mechanism in insects remains elusive. In the present work, immune-priming was detected in a lepidopteran insect, Spodoptera exigua. Specifically, a prior infection with a heat-killed pathogenic bacterium, Escherichia coli, led to increased survival upon the second infection of different pathogens. Plasma collected from larvae with the prior infection possessed the immune-priming factor(s) that significantly up-regulated cellular and humoral immune responses of naïve larvae. Our study also finds that variations in the timing of plasma collection for priming larvae resulted in distinct impacts on both cellular and humoral responses. However, when the active plasma exhibiting the immune-priming was heat-treated, it lost this priming activity, therefore suggesting that protein factor(s) play a role in this immune-priming. An immunofluorescence assay showed that the hemocytes collected from the immune-primed larvae highly reacted to a polyclonal antibody specific to a vertebrate lipocalin, apolipoprotein D (ApoD). Among 27 ApoD genes (Se-ApoD1 ∼ Se-ApoD27) of S. exigua, Se-ApoD3 was found to be highly induced during the immune-priming, in which it was shown to be expressed in hemocytes and fat body from a fluorescence in situ hybridization analysis. RNA interference of Se-ApoD3 expression significantly impaired the immune-priming of S. exigua larvae. Moreover, the inhibition of eicosanoid biosynthesis suppressed the immune-priming, in which treatment with a lipoxygenase (LOX) inhibitor-and not treatment with a cyclooxygenase inhibitor-suppressed immune-priming. Further, an addition of LOX product such as lipoxin A 4 or lipoxin B 4 significantly rescued the lost immune-priming activity. Taken together, these results suggest that a complex of ApoD3 and LOX product mediates the immune-priming activity of S. exigua., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The oxidation of the cellular membrane through lipid peroxidation (LPO) is linked to aging and disease. Despite the physiological importance, the chemical mechanisms underlying LPO and oxidative reactions in membranes in general remain incompletely understood, and challenges exist in translating LPO inhibitor efficacies from in vitro to in vivo. The complexity of LPO, including multiple oxidation reactions in complex membrane environments and the difficulty in quantifying reaction kinetics, underlies these difficulties. In this work, we developed a robust and straightforward method for quantifying the oxidation rate kinetics of fluorescent molecules and determined the oxidation kinetics of widely fluorophores used as indicators of membrane LPO, diphenylhexatriene (DPH), BODIPY-C11, and Liperfluo. The measurement is initiated by lipoxygenase, which provides chemical specificity and enables a straightforward interpretation of oxidation kinetics. Our results reveal that the membrane composition significantly impacts the observed kinetics oxidation in DPH and BODIPY-C11 but not Liperfluo. Reaction mechanisms for their lipid peroxide-induced oxidation are proposed. This work provides a foundation for the quantitative analysis of LPO with fluorescence and extricating the complexity of oxidation reactions within membranes.

Nicotinamide adenine dinucleotide (NAD + ) is a redox cofactor and signal central to cell metabolisms. Disrupting NAD homeostasis in plant alters growth and stress resistance, yet the underlying mechanisms remain largely unknown. Here, by combining genetics with multi-omics, we discover that NAD + deficiency in qs-2 caused by mutation in NAD + biosynthesis gene-Quinolinate Synthase retards growth but induces biosynthesis of defense compounds, notably aliphatic glucosinolates that confer insect resistance. The elevated defense in qs-2 is resulted from activated jasmonate biosynthesis, critically hydroperoxidation of α-linolenic acid by the 13-lipoxygenase (namely LOX2), which is escalated via the burst of chloroplastic ROS-singlet oxygen ( 1 O 2 ). The NAD + deficiency-mediated JA induction and defense priming sequence in plants is recapitulated upon insect infestation, suggesting such defense mechanism operates in plant stress response. Hence, NAD homeostasis is a pivotal metabolic checkpoint that may be manipulated to navigate plant growth and defense metabolism for stress acclimation., (© 2024. The Author(s).)

Traditional cancer chemotherapy suffers from low efficacy and severe side effects, limiting its use as a first-line treatment. To address this issue, we investigated a novel way to induce lipid peroxidation (LPO), which plays an essential role in ferroptosis and may be useful against cancer cells and tumors. In this study, a pH-responsive synergistic cancer therapy nanoplatform was prepared using CaCO 3 co-loaded with oleanolic acid (OA) and lipoxygenase (LOX), resulting in the formation OLCaP NP. This nanoplatform exhibited good drug release properties in an acidic tumor environment owing to the presence of CaCO 3 . As a result of acidic stimulation at tumor sites, the OLCaP NP released OA and LOX. OA, a chemotherapeutic drug with anticancer activity, is already known to promote the apoptosis of cancer cells, and LOX is a natural enzyme that catalyzes the oxidation of polyunsaturated fatty acids, leading to the accumulation of lipid peroxides and promoting the apoptosis of cancer cells. More importantly, OA upregulated the expression of acyl-coenzyme A synthetase long-chain family member 4 (ACSL4), which promoted enzyme-mediated LPO. Based on our combined chemotherapy and nanocatalytic therapy, the OLCaP NP not only had remarkable antitumor ability but also upregulated ACSL4 expression, allowing further amplification of LPO to inhibit tumor growth. These findings demonstrate the potential of this nanoplatform to enhance the therapeutic efficacy against tumors by inducing oxidative stress and disrupting lipid metabolism, highlighting its clinical potential for improved cancer treatment. STATEMENT OF SIGNIFICANCE: This study presents a novel nanoplatform that combines oleanolic acid (OA), a chemotherapeutic drug, and lipoxygenase (LOX), which oxidizes polyunsaturated fatty acids to trigger apoptosis, for targeted cancer therapy. Unlike traditional treatments, our nanoplatform exhibits pH-responsive drug release, specifically in acidic tumor environments. This innovation enhances the therapeutic effects of OA and LOX, upregulating acyl-CoA synthetase long-chain family member 4 expression and amplifying lipid peroxidation to promote tumor cell apoptosis. Our findings significantly advance the existing literature by demonstrating a synergistic approach that combines chemotherapy and nanocatalytic therapy. The scientific impact of this work lies in its potential to improve cancer treatment efficacy and specificity, offering a promising strategy for clinical applications and future research in cancer therapy., 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 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Menopause is a normal stage in the human female aging process characterized by the cessation of menstruation and the ovarian production of estrogen and progesterone hormones. Menopause is associated with an increased risk of several different diseases. Cardiovascular diseases are generally less common in females than in age-matched males. However, this female advantage is lost after menopause. Cardiac hypertrophy is a disease characterized by increased cardiac size that develops as a response to chronic overload or stress. Similar to other cardiovascular diseases, the risk of cardiac hypertrophy significantly increases after menopause. However, the exact underlying mechanisms are not yet fully elucidated. Several studies have shown that surgical or chemical induction of menopause in experimental animals is associated with cardiac hypertrophy, or aggravates cardiac hypertrophy induced by other stressors. Arachidonic acid (AA) released from the myocardial phospholipids is metabolized by cardiac cytochrome P450 (CYP), cyclooxygenase (COX), and lipoxygenase (LOX) enzymes to produce several eicosanoids. AA-metabolizing enzymes and their respective metabolites play an important role in the pathogenesis of cardiac hypertrophy. Menopause is associated with changes in the cardiovascular levels of CYP, COX, and LOX enzymes and the levels of their metabolites. It is possible that these changes might play a role in the increased risk of cardiac hypertrophy after menopause., Competing Interests: Declaration of Competing Interest The authors declare that there are no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Argemone mexicana belonging to family Papaveraceae is a traditional medicinal plant widely utilized by tribal people in India for treating various ailments like skin infections, wounds and inflammation. This plant is very rich in alkaloidal content, which has a great potential in the treatment of anti-inflammatory disorders. Therapeutically promising bioactive molecules are often produced by endophytic fungi associated with medicinal plants. In this investigation, endophytic fungi were isolated from various parts of A. mexicana and screened for alkaloidal content. Among these, one of the fungal isolate, Acremonium alternatum AMEF-5 producing maximum alkaloids showed significant anti-inflammatory activity. Fractionation of this crude fungal extract through column chromatography yielded eight fractions, which were further screened for anti-inflammatory activities. Fraction 3 exhibited significant anti-inflammatory activity by the inhibition of lipoxygenase enzyme (IC 50 15.2 ± 0.09 µg/ml), scavenging of the nitric oxide radicals (IC 50 11.38 ± 0.35 µg/ml), protein denaturation (IC 50 14.93 ± 0.4 µg/ml), trypsin inhibition (IC 50 12.06 ± 0.64 µg/ml) and HRBC stabilization (IC 50 11.9 ± 0.22 µg/ml). The bioactive alkaloid in fraction 3 was identified as aconitine which was confirmed by UV, FTIR, HPLC, HRMS, 1 H NMR, and 13 C NMR analysis. This study demonstrates that endophytic fungi serve a potential source for sustainable production of therapeutically important alkaloids., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

The oxidation of fish lipids and proteins is interconnected. The LOX (lipoxygenase)-catalyzed LA (linoleic acid) oxidation system on MPs (myofibrillar proteins) was established in vitro, to investigate the impact of lipoxidation on the physicochemical properties of fish MPs. By detecting HNE (4-hydroxy-2-nonenal) concentration during LA oxidation, the HNE treatment system was established to investigate the role of HNE in this process. In addition, the site specificity of modification on MPs was detected utilizing LC-MS/MS. Both treatments could induce sidechain modification, increase particle size, and cause loss of nutritional value through the reduction in amino acid content of MPs. The HNE group is more likely to alter the MPs' surface hydrophobicity compared to the LA group. By increasing the exposure of modification sites in MPs, the HNE group has more types and number of modifications compared to the LA group. LA group mainly induced the modification of single oxygen addition on MPs instead, which accounted for over 50 % of all modifications. The LA group induced a more pronounced reduction in the solubility of MPs as compared to the HNE group. In conclusion, HNE binding had a high susceptibility to Lys on MPs. Protein aggregation, peptide chain fragmentation, and decreased solubility occurred in the LA group mainly induced by peroxide generated during lipid oxidation or the unreacted LA instead of HNE. This study fills in the mechanism of lipoxidation on protein oxidation in fish and sheds light on the HNE modification sites of MPs, paving the way for the development of oxidation control technology., 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.)

This study aimed to explore the antioxidant and prooxidative activity of two natural furanocoumarin derivatives, Bergaptol (4-Hydroxy-7H-furo [3,2-g] [1]benzopyran-7-one, BER) and Xanthotoxol (9-Hydroxy-7H-furo [3,2-g] [1]benzopyran-7-one, XAN). The collected thermodynamic and kinetic data demonstrate that both compounds possess substantial antiradical activity against HO • and CCl 3 OO • radicals in physiological conditions. BER exhibited better antiradical activity in comparison to XAN, which can be attributed to the enhanced deprotonation caused by the positioning of the -OH group on the psoralen ring. In contrast to highly reactive radical species, newly formed radical species BER • and XAN • exhibited negligible reactivity towards the chosen constitutive elements of macromolecules (fatty acids, amino acids, nucleobases). Furthermore, in the presence of O 2 •─ , the ability to regenerate newly formed radicals BER • and XAN • was observed. Conversely, in physiological conditions in the presence of Cu(II) ions, both compounds exhibit prooxidative activity. Nevertheless, the prooxidative activity of both compounds is less prominent than their antioxidant activity. Furthermore, it has been demonstrated that anionic species can engage in the creation of a chelate complex, which restricts the reduction of metal ions when reducing agents are present (O 2 •─ and Asc ─ ). Moreover, studies have demonstrated that these chelating complexes can be coupled with other radical species, hence enhancing their ability to inactivate radicals. Both compounds exhibited substantial inhibitory effects against enzymes involved in the direct or indirect generation of ROS: Xanthine Oxidase (XOD), Lipoxygenase (LOX), Myeloperoxidase (MPO), NADPH oxidase (NOX)., 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.)

The synthesis, antioxidant capacity, and anti-inflammatory activity of four novel N -benzyl-2-[4-(aryl)-1 H -1,2,3-triazol-1-yl]ethan-1-imine oxides 10a - d are reported herein. The nitrones 10a - d were tested for their antioxidant properties and their ability to inhibit soybean lipoxygenase (LOX). Four diverse antioxidant tests were used for in vitro antioxidant assays, namely, interaction with the stable free radical DPPH (1,1-diphenyl-2-picrylhydrazyl radical) as well as with the water-soluble azo compound AAPH (2,2'-azobis(2-amidinopropane) dihydrochloride), competition with DMSO for hydroxyl radicals, and the scavenging of cationic radical ABTS •+ (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) radical cation). Nitrones 10b , 10c, and 10d , having the 4-fluorophenyl, 2,4-difluorophenyl, and 4-fluoro-3-methylphenyl motif, respectively, exhibited high interaction with DPPH (64.5-81% after 20 min; 79-96% after 60 min), whereas nitrone 10a with unfunctionalized phenyl group showed the lowest inhibitory potency (57% after 20 min, 78% after 60 min). Nitrones 10a and 10d , decorated with phenyl and 4-fluoro-3-methylphenyl motif, respectively, appeared the most potent inhibitors of lipid peroxidation. The results obtained from radical cation ABTS •+ were not significant, since all tested compounds 10a - d showed negligible activity (8-46%), much lower than Trolox (91%). Nitrone 10c , bearing the 2,4-difluorophenyl motif, was found to be the most potent LOX inhibitor (IC 50 = 10 μM).