Investigation of molecular and structural properties of two Mu-class GSTs from Tetranychus urticae.

Mining of key genes involved in the sulforaphane biosynthetic pathway of moringa and cloning, expression, and functional verification of MoMYR1.

Glucosinolates (GSLs) from cruciferous vegetables (CVs), sulfur (S)- and nitrogen-containing compounds, are enzymatically hydrolyzed by myrosinase (EC 3.2.1.147) to yield bioactive derivatives such as isothiocyanates (ITCs) and indoles. These metabolites exhibit chemopreventive and anticancer properties. The article compiles evidence regarding the following: (i) the molecular mechanisms regulating the biosynthesis of key derivatives, including sulforaphane (SFN), phenethyl isothiocyanate (PEITC), and indole-3-carbinol (I3C); (ii) epidemiological and clinical findings; and (iii) strategies to link plant science with nutritional interventions for cancer prevention. An integrative literature review was conducted using Web of Science, Scopus, ScienceDirect, Google Scholar, and PubMed. English-language studies addressing mechanistic insights, nutritional factors, epidemiology, and clinical trials were included. The biosynthesis and metabolism of GSL in plants are regulated by S and several transcription factors that promote or repress GSL production. Additionally, food processing has been shown to influence retention time and the formation of ITCs. In humans, ITCs activate nuclear factor erythroid 2-related factor 2 (Nrf2)-mediated detoxification, induce apoptosis, and modulate epigenetic pathways. Epidemiological data show inverse associations between CV intake and cancer risk, though variability exists. Clinical trials have confirmed the bioavailability and effects of glucoraphanin and SFN on cancer-related biomarkers. The described compounds are bioavailable in humans and modulate the clinically relevant pathways linked to carcinogenesis. Larger, standardized interventions are needed to determine effective intake levels, optimize bioavailability, and define their potential role in evidence-based nutritional strategies for cancer prevention.

Dietary isothiocyanates inhibit the oxidative activity of salivary aldehyde dehydrogenase ALDH3A1 and modulate aroma release.

Isothiocyanates (ITCs) are well-known electrophilic agents with antioxidant and anticancer properties, largely attributed to their ability to activate the Nrf2/ARE pathway. Building on previous work with C1-ITC glycosyl derivatives, we designed and synthesized a new series of S-glycosyl isothiocyanates in which the ITC group was repositioned to the C6 carbon of the glucose scaffold. This structural rearrangement yielded stable and synthetically accessible derivatives with markedly enhanced biological profiles. Several compounds showed potent Nrf2 activation at non-cytotoxic concentrations, with CD values comparable to or exceeding those of natural ITCs. In parallel, the new C6-ITC derivatives displayed significant antiproliferative activity against leukemia and solid tumor cell lines. Among them, the phenylsulfone derivative 13 emerged as a particularly promising dual-action molecule, combining strong Nrf2 induction with low-micromolar cytotoxicity. Molecular docking was used as a hypothesis-generating approach and suggested a possible interaction with the STAT3 SH2 domain, although further studies are needed to validate this target. Overall, these results support glucose-based ITCs as a versatile platform for the development of multifunctional antioxidants with complementary anticancer properties.

Our previous results demonstrated that depletion of glutathione (GSH) rather than elevation of levels of reactive oxygen species (ROS) is highly correlated with the decrease in stomatal aperture induced by isothiocyanates (ITCs), although ROS is considered a key second messenger in stomatal closure, suggesting that another signal component regulates stomatal apertures along with GSH depletion. This study, using Arabidopsis, clarified that reactive carbonyl species (RCS), especially acrolein and 4-hydroxy-(E)-2-nonenal, are determinants of stomatal aperture responses to ITCs. All tested ITCs, allyl isothiocyanate (AITC), sulforaphane (SFN), benzyl isothiocyanate (BITC), and phenethyl isothiocyanate (PEITC), significantly induced stomatal closure, which was inhibited by the RCS scavengers, carnosine and pyridoxamine. The RCS scavengers suppressed ITC-induced depletion of GSH but not elevation of ROS levels. All tested ITCs (AITC, SFN, BITC, and PEITC) increased levels of RCS and non-RCS aldehydes in the epidermal tissues. However, acrolein, 4-hydroxy-(E)-2-nonenal, crotonaldehyde, and (E)-2-pentenal induced stomatal closure at 10 and 100 μM, whereas propionaldehyde, butyraldehyde, and n-pentanal did not at concentrations up to 100 μM. Acrolein and 4-hydroxy-(E)-2-nonenal more effectively induced stomatal closure and GSH depletion than crotonaldehyde and (E)-2-pentenal did. The contents of RCS were more strongly correlated with GSH levels and stomatal closure than with ROS levels. These results suggest that RCS, especially acrolein and 4-hydroxy-(E)-2-nonenal, acts as key regulators of stomatal closure in guard cells in response to ITCs.

The rape stem weevil (Ceutorhynchus asper) represents a significant threat to economically-important cruciferous crops. The main aim was to characterize CaspOBPs (odorant-binding proteins) and the functions of CaspOBP9. We performed transcriptomic sequencing on C. asper tissues and identified 21 CaspOBPs. A majority of CaspOBPs were relatively highly expressed in the antennae; for these OBPs, CaspOBP9, 12, 16, 19 and 20 were female-biased, whereas CaspOBP5, 8 and 10 were male-biased. Recombinant CaspOBP9 showed good affinity for four plant volatiles and two pesticides, suggesting that CaspOBP9 was involved in perception of both pesticides and host volatiles. Behavioral experiments showed that C. asper was significantly attracted to decanal, benzyl isothiocyanate (ITC) and phenylethyl isothiocyanate. Molecular simulations suggest that Van der Waals forces serve as the main driving force for the formation of CaspOBP9-ligand complexes. Residues LEU:71, THR:79 and TYR:105 were identified to be critical in the formation of the complexes. Site-directed mutagenesis experiments showed that mutated proteins at the above-mentioned three sites had reduced or even no binding affinity for the corresponding compounds. RNAi experiments targeting CaspOBP9 confirmed its essential roles in olfactory perception of C. asper. CaspOBP9 could play important roles in perception of particular plant volatiles (e.g. benzyl ITC) and pesticides (e.g. chlorpyrifos) by C. asper, and its key residues LEU:71, THR:79 and TYR:105 were identified. This research provides valuable insights into the molecular mechanisms underlying olfactory perception of C. asper, and a foundation for the development of novel control techniques for this weevil. © 2025 Society of Chemical Industry.

Brassica oleracea vegetables (e. g. cabbages) form bioactive isothiocyanates (ITCs) from glucosinolate (GLS) hydrolysis. However, enzymatic activity, acidic pH (below pH 5), and ferrous ions (Fe2+) can promote nitrile release, reducing the ITC amount. In Arabidopsis thaliana, nitrile-specifier proteins (NSPs) promote nitrile formation upon GLS hydrolysis. Here, we report the functional characterization of two Brassica NSPs from B. oleracea and the in silico identification of candidate genes encoding a family of sixteen B. oleracea NSPs closely related to the A. thaliana NSPs and the likely ancestral protein, XP_013585314.1. High conservation of the iron-binding triad (EXXXDXXXH), characteristic of specifier proteins, was confirmed in the putative BoNSPs. Biochemical characterization of two B. oleracea NSP isoforms, BoNSP2 (XP_013609641.1) and BoNSP11 (XP_013587057.1), revealed increased NSP activity in the presence of added Fe2+. Both BoNSP isoforms affected hydrolysis of five GLS differently in vitro, suggesting differential substrate specificity. BoNSP2 showed higher nitrile formation from indol-3-ylmethyl GLS than from 4-(methylsulfinyl)butyl GLS. In contrast, BoNSP11 similarly increased nitrile formation from indol-3-ylmethyl GLS, three aliphatic GLS and benzyl GLS. BoNSP2 and BoNSP11 were most active between pH 7 and pH 8. This study identifies and characterizes the first NSPs in B. oleracea vegetables at the molecular level.

Synergistic strategies of sulforaphane biosynthesis and functional properties in broccoli sprouts powder obtained from sucrose stress modulation coupled with vacuum freeze-drying.

Breast cancer (BC) is a leading cause of cancer-related mortality, with estrogen receptor (ER)-negative subtypes, especially triple-negative BC, comprising one-fifth of global cases. Natural inhibitors, particularly those from cruciferous vegetables like arugula (Eruca sativa), which are rich in bioactive isothiocyanates (ITCs), show potent anticancer effects and cytoprotection when combined with chemotherapy. Sulforaphane (SFN) and its analogue erucin modulate oxidative stress, detoxification, and epigenetic pathways. This study computationally assessed their anti-cancer potential in ER-negative BC using transcriptomic analysis, molecular docking, and ADMET profiling. Microarray data (GSE28813) from SFN-treated ER-negative MCF10A cells were analyzed via GEO2R and GEOExplorer to identify highly upregulated differentially expressed genes (DEGs). Key DEGs included AKR1B10 (logFC = 7.26), AKR1C1 (logFC = 5.10), AKR1C3 (logFC = 4.42), NMRAL1P1 (logFC = 6.42), and HKDC1 (logFC = 6.13). Elevated AKR1B10 is strongly linked to early BC malignancies, positioning it as a diagnostic and therapeutic target. Molecular docking showed SFN's superior binding affinity to AKR1B10 compared to erucin, with strong interactions at catalytic site residues via hydrogen and hydrophobic bonds. ADMET profiling confirmed SFN's high intestinal absorption and blood-brain barrier non-permeability. Thus, integrating SFN as a natural AKR1B10 inhibitor into ER-negative BC treatment regimens may enhance early malignancy management and support its development as a nutraceutical adjunct.

Cabbage (Brassica oleracea var. capitata) contains glucosinolates (GSLs) such as glucoraphanin and sinigrin, which serve as precursors to bioactive isothiocyanates (ITCs), including sulforaphane (SFN) and allyl isothiocyanate (AITC). Upon cellular rupture, GSLs come into contact with myrosinase, resulting in ITC formation as part of the plant defense response. While this process has been studied biochemically, its spatial manifestation within plant tissues has not been directly visualized. In this study, GSL levels in intact and ruptured cabbage samples were quantified by LC-MS, and imaging mass spectrometry (IMS) was used to map the GSLs and ITCs in tissue sections. GSLs were localized mainly in intact regions, whereas ITCs were detected predominantly in ruptured areas, showing reciprocal spatial distributions. These results provide the first direct visualization of GSL-to-ITC conversion in plant tissues and demonstrate the utility of IMS for studying enzymatic biochemical transformations relevant to plant defense and food functionality.

The rising global prevalence of obesity and metabolic disorders calls for innovative dietary strategies that can modulate key enzymatic pathways involved in lipid and carbohydrate metabolism. This study uncovers the effects of sulforaphane (SFN)-rich broccoli-derived formulations—including liquid and lyophilised forms, as well as two commercial prototypes, Sulforaphan® BASIC and Sulforaphan® SMART, the latter being characterised by the inclusion of an enteric-coated myrosinase enzyme designed to enhance the in situ conversion of glucosinolates (GSL) into bioactive isothiocyanates (ITC)—on lipid and carbohydrate metabolism in 3T3-L1 adipocytes. Across the formulations, total GSL content ranged widely, with GS0 showing the highest levels. Functionally, all SFN-rich formulations significantly reduced intracellular triglyceride content, with the SMART formulation achieving the strongest reduction (11% compared with untreated controls). Across enzymatic assays, we recorded that every formulation inhibited lipoprotein lipase and α-glucosidase activities, with Sulforaphan® BASIC and Sulforaphan® SMART leading to moderate inhibition (40–50%). The potent effect of SMART formulation may be associated with the presence of enteric-coated myrosinase, which enhances the conversion of GSL into bioactive ITC. The gathered evidence provides further insights into the potential of bioactive compounds in cruciferous foods to modulate metabolic health, underscoring their potential role in complementary therapeutic strategies for obesity and its comorbidities.

In this article, the granules based on polycaprolactone (PCL) and bentonite in the ratio of 75/25 and 50/50 were obtained as a promising carrier for long-term release of isothiocyanates (ITC). According to GC-MS and FTIR spectroscopy, it was found that concentration of ITC in granules increases with increase in clay ratio and depends on the ITC species. It is observed that the inclusion of ITC in PCL/bentonite granules increases the persistence of ITCs in the soil for up to 45 days and prevents their premature hydrolysis.

The conversion of glucosinolates (GSLs) into chemopreventive isothiocyanates (ITCs) primarily relies on plant myrosinase (MYR) or specific bacteria. MYR dynamics are deeply involved in plant defense systems, gut microbiota metabolism, and complex interactions and regulation across species. A set of activity-based probes was developed to track MYR in vivo by biomimicking natural GSL with robust sensitivity and selectivity. The dynamics and heterogeneous distribution of MYR in distinct sections and species were captured via fluorescence imaging of live plants. Specifically, under herbivore challenge to leaves, a systemic, long-distance upregulation of MYR activity in root tissues has confirmed cross-species MYR regulation in plant defense. Furthermore, for the first time, quantitative visualization of the dynamic metabolic competition of GSL and sugar has confirmed the metabolic priority of sugar in gut microbiota and colonized zebrafish in vivo. The competitive metabolism is involved in the crosstalk during cross-species microbes and host-microbe interactions. Tracking MYR regulation across species by the designed probes has offered rich insights into the dynamic interplay among diet, microbiota, and host health.

Heteroatom incorporation is a general strategy toward advanced polymer properties. We report here efficient synthesis of polymers containing, site-specifically, at least four different heteroatoms (O, N, S, F) by copolymerization of isothiocyanate (ITC) and trifluoropropylene oxide (TFPO). Unlike the regular non-fluorinated epoxides for which Lewis pair catalysts are needed, the superior activity of TFPO allows the use of only organobases, even the relatively mild ones, to reach high/complete monomer conversion in hours at room temperature and low catalyst loadings (0.01-0.1 mol.%). With optimal initiator structure and monomer feed ratio (1/1), alternating copolymers, i.e. fluorinated polythioimidocarbonates (FPTCs), with controlled molar mass and low dispersity are obtained. Copolymerization with alkyl and allyl ITCs are remarkably more chemoselective, which is ascribed to the alleviated nucleophilicity of the oxyanionic species and expedited S-to-O crossover. While thermostability of the aromatic PTC is evidently improved and adhesive strength unaffected by the CF3 groups, depolymerization becomes much easier under basic conditions, selectively forming a five-membered cyclic thioimidocarbonate.

Background: Nasturtium (Tropaeolum majus L.), an edible flower originating from the Andean region of South America, is rich in phenolic compounds derived from quinic acid, flavonoids, glucosinolates (GLS), and their secondary metabolites, isothiocyanates (ITCs), which possess antioxidant and anti-inflammatory effects. Objective: This study evaluated the impact of sub-chronic consumption of a freeze-dried nasturtium leaves drink on inflammatory and antioxidant biomarkers in individuals diagnosed with prediabetes. Methods: In this randomized, crossover trial, 10 prediabetic adults (aged 25-70 years) received either nasturtium (NT) or placebo (PLC) for 4 weeks, then crossed over to the other treatment for another 4 weeks. Blood samples before and after each treatment were analyzed for gene expression related to inflammation and antioxidant response, and total antioxidant capacity (TAC) was measured using ORAC and ABTS assays, along with oxidized LDL (ox-LDL) levels. Results: Weekly consumption of 15 g NT for four weeks significantly increased TAC by 21% (ORAC) and 10% (ABTS), and reduced ox-LDL by 13%. No changes in gene expression were observed. Novelty of the Study: This study is the first to demonstrate that consuming nasturtium specifically increases TAC and decreases ox-LDL in prediabetic patients. The results underscore the antioxidant potential of nasturtium as a functional food or nutraceutical to reduce oxidative stress-driven progression toward type 2 diabetes. Conclusions: Nasturtium intake enhances antioxidant capacity and may help reduce oxidative stress associated with the development of type 2 diabetes. Further research is needed to confirm its role in T2D prevention. Trial registration: NCT05346978, dated 23 April 2022. Keywords: Pre-diabetes, Tropaeolum majus L., Glucotropaeolin, BITC, Total antioxidant capacity, ox-LDL

Eruca sativa Mill. (rocket; Fam. Brassicaceae) is widely appreciated for its peculiar flavour and beneficial effects on human health. Glucosinolates (GSLs) and their enzymatic hydrolysis products, isothiocyanates (ITCs), are considered to be responsible for health-promoting effects and for sensory relevance in rocket, respectively. This study aimed at evaluating and comparing the metabolite profiles of rocket leaves collected at different phenological stages, to investigate the content evolution during cultivation. To minimise metabolic variability induced by environmental factors, plants were cultivated in an innovative growing system equipped with precision lighting and ventilation. A multi-platform metabolomics approach combining liquid chromatography–high-resolution mass spectrometry (LC–HRMS) and headspace solid-phase microextraction coupled with gas chromatography–mass spectrometry (HS-SPME/GC–MS) was carried out for comprehensive coverage of non-volatile and volatile organic compounds (VOCs). To integrate data from both platforms, a multivariate data fusion strategy was used. Higher GSLs content was detected in the microgreens stage. In particular, glucoraphanin, glucoiberverin, glucoerucin, DMB-GLS, and 1,4-dimethoxyglucobrassicin were identified as biological markers of rocket microgreens. ITCs levels were found to increase in mature leaves. These findings suggest a dynamic modulation of secondary metabolism during the plant life cycle, possibly in response to different adaptation needs to environmental conditions. Our findings confirm the potential of microgreens as a functional food in promoting health and preventing chronic diseases and can also tailor rocket cultivation to maximise the production of beneficial metabolites and to improve selected sensorial features.

A simple waste-to-wealth strategy: sustainable bioconversion of rapeseed meal-derived glucosinolates into antimicrobial isothiocyanates.

Abstract Cyclic peptides represent a very useful modality in modern drug discovery. However, there remain substantial technical challenges in the de novo biosynthesis of complex cyclic peptides particularly in an intracellular manner. Herein, we demonstrate that an isothiocyanate (ITC) group encoded via genetic code expansion could selectively and efficiently crosslink to proximal N‐terminal Pro (P‐ITC crosslinking) and affords cyclic peptides without obvious limitations related to amino acid composition (including Lys and Cys) or sequence length. Notably via P‐ITC crosslinking, a disulfide‐bridged 13‐mer bicyclic peptide is facilely constructed in E. coli , thus lending us the ability to intracellularly construct and select complex cyclic peptides.

Cyclic peptides represent a very useful modality in modern drug discovery. However, there remain substantial technical challenges in the de novo biosynthesis of complex cyclic peptides particularly in an intracellular manner. Herein, we demonstrate that an isothiocyanate (ITC) group encoded via genetic code expansion could selectively and efficiently crosslink to proximal N-terminal Pro (P-ITC crosslinking) and affords cyclic peptides without obvious limitations related to amino acid composition (including Lys and Cys) or sequence length. Notably via P-ITC crosslinking, a disulfide-bridged 13-mer bicyclic peptide is facilely constructed in E. coli, thus lending us the ability to intracellularly construct and select complex cyclic peptides.