Plant Secondary Metabolites Research Articles

Variations in floral secondary metabolites of three flower color morphs of Lantana camara L.: possible roles in pollinators attraction

Plant secondary metabolites in the flowers mediate their interactions with pollinators. We analyzed the variations in the floral secondary metabolites (FSMs) of three different flower color morphs (FCMs) of Lantana camara in relation to pollinator attraction. FSMs were characterized by GC–MS. Data analysis revealed no significant quantitative differences between the FCMs in the chemical classes and insect semiochemicals of the FSMs. The possible influence of the FSMs acting as semiochemicals for the recorded pollinators, comprising butterflies (n = 16), bees (n = 3), and one moth species, based on the qualitative differences in the insect semiochemicals among the FCMs, was indicated. However, the correlation between the dominant pollinator abundance and the number of related semiochemicals was not found to be significant. Since the primary visual cues for butterflies are flower colors, we concluded that the insect semiochemicals of FSMs may play a secondary role in attracting butterflies. In contrast, the other FSMs may play a primary role in attracting bees and moth species. The reasons behind the conclusions are discussed based on the collected data.

Chemoproteomic Profiling Reveals Chlorogenic Acid as a Covalent Inhibitor of Arabidopsis Dehydroascorbate Reductase 1.

Chlorogenic acid (CA) is an abundant plant secondary metabolite with promising allelopathic effects on weed growth. However, the molecular targets and mechanism of action of CA in plants remain elusive. Here, we report the employment of a clickable photoaffinity probe in identifying the protein targets of CA in Arabidopsis seedling proteomes. CA specifically binds Arabidopsis dehydroascorbate reductase 1 (AtDHAR1), an enzyme responsible for ascorbate regeneration in plants, by covalent alkylating Cys20 within the catalytic center, thereby inhibiting its activity. In vivo application of CA reduced the pool size and redox state of ascorbate, leading to H2O2 accumulation in Arabidopsis seedlings. In agreement with these results, CA significantly induced the upregulation of antioxidant enzymes and downregulation of proteins involved in water transport and photosynthesis, as evidenced by quantitative proteomics. Taken together, this study revealed DHAR1 as a functional target underlying CA's allelopathic activity in plants, which opens new opportunities for the development of novel herbicides from naturally existing resources.

Integrated Transcriptomic and Metabolomic Analysis Revealed Regulatory Mechanisms on Flavonoids Biosynthesis in the Skin of Passion Fruit (Passiflora spp.).

Passion fruit is one of the most famous fruit crops in tropical and subtropical regions due to its high edible, medicinal, and ornamental value. Flavonoids, a class of plant secondary metabolites, have important health-related roles. In this study, a total of 151 flavonoid metabolites were identified, of which 25 key metabolites may be the main contributors to the purple phenotype. Using RNA sequencing, 11,180 differentially expressed genes (DEGs) were identified. Among these, 48 flavonoid biosynthesis genes (PAL, 4CL, C4H, CHS, CHI, F3H, DFR, ANS, and UFGT) and 123 transcription factors were identified. Furthermore, 12 distinct modules were identified through weighted gene coexpression network analysis, of which the brown module displays a robust positive correlation with numerous flavonoid metabolites. Overexpression of PeMYB114 significantly promoted flavonoids accumulation in tobacco leaves. Our study provided a key candidate gene for molecular breeding to improve color traits in passion fruit.

In silico molecular docking and molecular dynamic simulation of agarwood compounds with molecular targets of Alzheimer’s disease

Background Alzheimer’s disease (AD) is a neurological condition that primarily affects older people. Currently available AD drugs are associated with side effects and there is a need to develop natural drugs from plants. Aquilaria is as an endangered medicinal plant genus (commonly called agarwood plants) and various products of Aquilaria plant spp. including resinous heartwood, leaves, bark, and stem have been widely used in various traditional medicine systems. Research on agarwood plants is sparse and only a few previous studies demonstrated their neuroprotective properties in vitro. Owing to the presence of a plethora of secondary metabolites in agarwood plants, it is imperative not only to protect these plants but also evaluate the bioactivity of agarwood phytochemicals. Methods This study used Molsoft tools to predict the physicochemical properties of agarwood ligands, including the number of H-bond donors and acceptors, polar surface area, lipophilicity, solubility, and the molecular polar surface area and volume of agarwood ligands. Additionally, ADMET (absorption, digestion, metabolism, excretion, and toxicity) properties were predicted using ADMETlab 2.0. Computational methods such as AutoDock Vina and molecular dynamic (MD) simulations were employed for the docking of 41 selected agarwood compounds with AD-related molecular targets. Results and Conclusion According to docking data, three compounds aquilarisin (ASN), aquilarisinin (ANN), aquilarixanthone (AXN) showed highest binding affinity to selected AD targets compared to their known inhibitors. MD simulation studies revealed that, selected agarwood compounds’ protein-ligand complexes showed remarkable structural stability throughout 100ns simulation. The agarwood chemicals aquilarisin, aquilarisinin, aquilarixanthone, pillion (PLN), and agarotetrol (AGT) are consequently suggested as some of the found hits against AD targets, however, additional experimental validation is required to establish their effectiveness.

Bioconversion of Food and Green Waste into Valuable Compounds Using Solid-State Fermentation in Nonsterile Conditions.

Agro-industrial residues have transitions from being an environmental problem to being a cost-effective source of biopolymers and value-added chemicals. However, the efficient extraction of the desired products from these residues requires pretreatments. Fungal biorefinery is a fascinating approach for the biotransformation of raw materials into multiple products in a single batch. In this study, the ability of Trichoderma asperellum R to convert fruit scrap and green waste into value-added chemicals was tested in solid-state and in nonsterile conditions. A solid-state fermentation protocol for a tray bioreactor was developed using spawn as the inoculum for nonsterile substrates. T. asperellum R drove the fermentation of both substrates, shaping the metabolites that were enriched in the secondary plant metabolites. Strain R showed cellulase activity only when inoculated on fruit scraps, resulting in increased amounts of polysaccharides in the crude extract. This extract was also enriched in vanillic acid and limonoid, which are intriguing compounds due to the increasing interest in their potential as biological nitrification inhibitors or food additives. Finally, trimethoxybenzaldehyde, an interesting chemical building block, was identified in the extracts of the Trichoderma-guided fermentation. The overall results showed that the application of T. asperellum R has potential as a driver to facilitate the extraction of bioactive substances from nonsterile recalcitrant substrates.

Draft genome of a Bacillus subtilis ICS isolated from an okra (Abelmoschus esculentus) extract exhibited genes related to plant growth-promoting potential.

Here, we report the draft genome of Bacillus subtilis ICS isolated from the aqueous extract of Abelmoschus esculentus. The isolate has a genome size of 4.15 Mb, 43.07% GC, 99.8% BUSCO completeness, and 91.15% CheckM completeness. The genome exhibited genes related to plant growth promotion and secondary metabolite production.

MYB proteins: Versatile regulators of plant development, stress responses, and secondary metabolite biosynthetic pathways.

MYB proteins are ubiquitous in nature, regulating key aspects of plant growth and development. Although MYB proteins are known for regulating genes involved in secondary metabolite biosynthesis, particularly phenylpropanoids, their roles in terpenoid, glucosinolate, and alkaloid biosynthesis remain less understood. This review explores the structural and functional differences between activator and repressor MYB proteins along with their roles in plant growth, development, stress responses, and secondary metabolite production. MYB proteins serve as central hubs in protein-protein interaction networks that regulate expression of numerous genes involved in the adaptation of plants to varying environmental conditions. Thus, we also highlight key interacting partners of MYB proteins and their roles in these adaptation mechanisms. We further discuss the mechanisms regulating MYB proteins, including autoregulation, epigenetics, and post-transcriptional and post-translational modifications. Overall, we propose MYB proteins as versatile regulators for improving plant traits, stress responses, and secondary metabolite production.

Comparative analysis of retrosynthesis applications for predicting of pathways for plant Secondary metabolite production

Secondary metabolites (SMs), organic compounds synthesized by plants, play crucial roles in their own physiology and within their ecological niches, and have extensive usages in industries like pharmaceuticals, cosmetics, and food. Due to the complex nature of these metabolites, utilizing microorganisms has been proposed for their efficient and cost-effective production. In addressing challenges in SM synthesis, retrosynthesis has become essential. Applications such as RetroPath2.0, BioNavi-NP, and RetroBioCat have been developed to predict and design biosynthetic pathways, supporting synthetic biology and metabolic engineering efforts. These applications employ different methodologies to enhance the synthesis of target compounds, yet often face limitations in user-friendliness, functionality, and adaptability. This study evaluates the potential of RetroPath2.0, RetroBioCat, and BioNavi-NP in predicting the production pathways of 11 alkaloids, a class of plant secondary metabolites (PSMs). Through comparative analysis, the efficacy of these applications in proposing alternative production strategies was assessed. Findings highlighted RetroPath2.0’s capability in outlining precise production pathways in host organisms, despite some restrictions in its broader practicability. The study also demonstrated the production pathways of dimethyltryptamine, nicotine, and higenamine in non-plant hosts, illustrating the practical uses of these applications.

A Comparative Transcriptome Analysis Revealing the Mechanism Underlying the Effect of Maternal Plant Age on Adventitious Rooting of Euryodendron excelsum Cuttings

Euryodendron excelsum H. T. Chang, a monotypic plant endemic to China, exhibits significant variations in adventitious root formation in cuttings across different age groups through unclear molecular mechanisms. In this study, we performed RNA-sequencing (RNA-seq) and data analysis at three stages—initial excision (S0), root primordia (S1), and emergence of adventitious roots (S2)—for lateral buds from transplanted triennial tissue culture seedlings and decennial E. excelsum plants. We identified 13,424 differentially expressed genes (DEGs) between different rooting stages and age groups, including 8216 upregulated and 5208 downregulated genes. A Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis indicated that the S1 and S2 DEGs identified were mainly involved in phenylpropanoid biosynthesis, plant hormone signal transduction, plant–pathogen interactions, biosynthesizing various secondary plant metabolites, and starch and sucrose metabolism. Key auxin-pathway genes were expressed at significantly higher levels in culture seedlings than in adult plants, as were WUSCHEL-related homeobox and LATERAL ORGAN BOUNDARIES DOMAIN genes (related to adventitial root growth and development), which together promote the formation, growth, and development of adventitial root primordia. The dysregulated expression of 15 DEGs identified by RNA-seq were verified via real-time quantitative polymerase chain reaction analysis. This study investigates the morphological processes of adventitious root formation in cuttings of E. excelsum of different ages, and the potential molecular mechanisms involved. Our finding provides a basis for research on cutting propagation, conservation applications, and optimization of tissue culture for E. excelsum, and may help improve the rooting rate of E. excelsum cuttings.

Exploring Noncentrifugal Sugar as a Partial Replacement for White Sugar in Low Methoxyl Pectin Confectionery Gels: Impacts on Physical and Rheological Properties.

Noncentrifugal sugar (NCS) is an unrefined, dark brown sugar containing minerals and plant secondary metabolites, unlike refined white sugar (WS). This study explored using NCS in confectionary jellies as an alternative sugar. We used different concentrations of NCS and WS to prepare low methoxyl pectin (LMP) confectionery gels characterized by their physical and rheological properties along with time-domain nuclear magnetic resonance (TD-NMR) relaxometry. The strongest LMP gel, with a hardness of 0.94 N, was achieved by substituting 25% of WS with NCS at a low CaCl2 concentration (0.075 M). Gels with up to 50% WS replaced by NCS showed comparable hardness to standard LMP gels made solely with WS at a 0.15 M CaCl2 concentration, attributed to NCS's unique constituents. The NCS-WS gel exhibited the shortest T2 values (139.8 ms) and self-diffusion coefficient values (4.99 × 10-10 m2/s), indicating a denser, more cross-linked structure that restricted water mobility. These findings suggest NCS's complex role in affecting LMP gels' chemical and physical properties, highlighting its potential as a partial WS replacement in LMP gelation-based products, with an additional source of minerals and antioxidants.

BBL enzymes exhibit enantiospecific preferences in the biosynthesis of pyridine alkaloids in Nicotiana tabacum L.

Plant species can accumulate secondary metabolites in optically pure form or, occasionally, as enantiomeric mixtures. Interestingly, enantiomers of the same molecule can confer different biological activities. In tobacco (Nicotiana tabacum L.), the pyridine alkaloids nicotine, nornicotine, anatabine, and anabasine naturally exist as scalemic mixtures of (R)- or (S)-enantiomers, with the (S)-isoforms predominating. The mechanisms by which tobacco alkaloid enantiomers accumulate remain largely unknown. Experiments were carried out involving tobacco genotypes possessing induced deleterious mutations in three genes coding for nicotine demethylase (NND) enzymes and three genes coding for Berberine Bridge Like (BBL) enzymes that act near the end of the nicotine, anatabine, and anabasine biosynthetic pathways. Data indicate that (R)-nicotine is naturally produced at appreciable levels but is observed in only small amounts due to preferential demethylation by NND enzymes. Data further suggest that BBL-a and BBL-b are preferentially involved in the biosynthesis of (S)-alkaloid enantiomers, while BBL-c is preferentially involved in the biosynthesis of (R)-enantiomers. Gene duplication followed by genetic divergence thus played a role in the evolution of scalemic alkaloid accumulation in tobacco. Through a combination of mutation breeding and transgene overexpression, tobacco genotypes were generated in which the predominant alkaloid enantiomers were reversed from the (S)- to the (R)-isoforms. These results shed light on the genetic control of alkaloid accumulation in N. tabacum and on mechanisms of scalemic mixture formation of secondary metabolites in plants.

Design strategies of polysaccharide, protein and lipid-based nano-delivery systems in improving the bioavailability of polyphenols and regulating gut homeostasis.

Polyphenols are plant secondary metabolites that have attracted much attention due to their anti-inflammatory, antioxidant, and gut homeostasis promoting effects. However, food matrix interaction, poor solubility, and strong digestion and metabolism of polyphenols cause barriers to their absorption in the gastrointestinal tract, which further reduces bioavailability and limits polyphenols' application in the food industry. Nano-delivery systems composed of biocompatible macromolecules (polysaccharides, proteins and lipids) are an effective way to improve the bioavailability of polyphenols. Therefore, this review introduces the construction of biopolymer-based nano-delivery systems and their application in polyphenols, with emphasis on improving the solubility, stability, sustained release and intestinal targeting of polyphenols. In addition, there are possible positive effects of polyphenol-loaded nano-delivery systems on modulating gut microbiota and gut homeostasis, with particular emphasis on modulating intestinal inflammation, metabolic syndrome, and gut-brain axis. It is worth noting that the safety of bio-based nano-delivery systems still need to be further studied. In summary, the application of the bio-based nano-delivery system to deliver polyphenols provides insights for improving the bioavailability of polyphenols and for the treatment of potential diseases in the future.

Molecular Activities and Mechanisms of Action of Substances and Molecules from Medicinal Plants from Sub-Saharan Africa on Prostate and Cervical Cancer Cells.

Despite years of medical research, cancer remains a major public health problem worldwide, particularly in Africa. The cost, duration, and toxicity of currently available treatments are all drawbacks. Plant secondary metabolites are significant anticancer compounds. Already used in traditional health systems, plants are currently the subject of numerous studies to discover new anti-cancer drugs. This review assesses the literature on the cytotoxic effect of plant substances (extracts) and molecules on prostate and cervical cancer cell lines. PubMed, Science Direct, and Google Scholar were used to find in vitro studies carried out between 2006 and 2023 related to the cytotoxicity of extracts, substances and/or molecules from plants harvested in sub- Saharan Africa against prostate and/or cervical cancer cell lines. A total of 36 reports on the cytotoxic potential of 96 medicinal plants from sub-Saharan Africa were extracted from the selected databases. All the plants listed had a cytotoxic effect on prostate and/or cervical cancer cells. Some plant extracts or molecules showed significant activity with an IC50< 20 μg/ml. Burkina Faso and South Africa had the most plant extracts tested for prostate and cervical cancer, respectively. A total of 36 reports on the cytotoxic potential of 96 medicinal plants from sub-Saharan Africa were extracted from the selected databases.

Subcritical Water as a Tool for Obtaining Extracts of Plant Secondary Metabolites with High Antioxidant Activity from Ginkgo biloba L. Leaves

Subcritical Water as a Tool for Obtaining Extracts of Plant Secondary Metabolites with High Antioxidant Activity from Ginkgo biloba L. Leaves

Naringenin Nanoformulations for Neurodegenerative Diseases.

Glioblastoma (GBM) is a grade-IV astrocytoma, which is the most common and aggressive type of brain tumor, spreads rapidly and has a life-threatening catastrophic effect. GBM mostly occurs in adults with an average survival time of 15 to 18 months, and the overall mortality rate is 5%. Significant invasion and drug resistance activity cause the poor diagnosis of GBM. Naringenin (NRG) is a plant secondary metabolite byproduct of the flavanone subgroup. NRG can cross the blood-brain barrier and deliver drugs into the central nervous system when conjugated with appropriate nanocarriers to overcome the challenges associated with gliomas through naringenin-loaded nanoformulations. Here, we discuss several nanocarriers employed that are as delivery systems, such as polymeric nanoparticles, micelles, liposomes, solid lipid nanoparticles (SLNs), nanosuspensions, and nanoemulsions. These naringenin-loaded nanoformulations have been tested in various in vitro and in vivo models as a potential treatment for brain disorders. This review nanoformulations of NRG can a possible therapeutic alternative for the treatment of neurological diseases are discussed.

Accumulation of secondary metabolites in the family Lamiaceae as influenced by foliar micronutrients

The Lamiaceae family is rich in secondary metabolites, such as essential oils comprising flavonoids, phenolic compounds, and terpenoids. Micronutrients play a crucial role in plant metabolism and secondary metabolites production. This review summarizes and emphasizes the significance of foliar micronutrient nutrition in facilitating the accumulation of secondary metabolites in the Lamiaceae family, based on research literature from 2010 to 2023 using dependable databases like Scopus, Google Scholar, PubMed, and Science Direct. Spraying micronutrients notably enhanced the biosynthesis of various secondary metabolites in Lamiaceae species, especially for nutrients that do not move easily (Fe, Zn, Mn, Cu, B, and Si). Using nano-micronutrients as foliar application on Lamiaceae medicinal plants was more effective than traditional soil treatment. This review highlighted the importance of additional investigation on Cerium, Titanium, and Boron. Genera such as Lavandula, Satureja, Origanum, Lippia, and Hyssopus require more research in the future.

Nitrilases NIT1/2/3 Positively Regulate Resistance to Pseudomonas syringae pv. tomato DC3000 Through Glucosinolate Metabolism in Arabidopsis.

Nitrilases, found to have a common presence in the plant kingdom, are capable of converting nitriles into their corresponding carboxylic acids through hydrolysis. In Arabidopsis, the nitrilases NIT1, NIT2, and NIT3 catalyze the formation of indole-3-acetonitrile (IAN) into indole-3-acetic acid (IAA). Notably, IAN can originate from the breakdown products of indole glucosinolates. Glucosinolates, which are plant secondary metabolites commonly found in cruciferous plants, and their breakdown products, are crucial for plant defense against pathogens. In our study, we found that nitrilases positively regulate resistance to Pseudomonas syringae pv. tomato DC3000 (PstDC3000) in mature Arabidopsis. Transcriptome data showed that after PstDC3000 treatment, genes related to the auxin pathway in nit1nit2nit3 changed more dramatically than in the wild type. Moreover, the enhancement of disease resistance through exogenous aliphatic glucosinolate application relies on NIT1/2/3. Hence, it is hypothesized that NIT1/2/3 may serve a dual role in disease resistance and defense mechanisms. After infection with PstDC3000, NIT1/2/3 catalyzes the biosynthesis of auxin, thereby triggering certain disease-related responses. On the other hand, NIT1/2/3 can also break down nitriles generated from aliphatic glucosinolate degradation to enhance disease resistance. Our study elucidates the regulatory mechanism of nitrilases in Arabidopsis disease resistance, offering a theoretical foundation for enhancing disease resistance in cruciferous plants.

Green synthesis of Brassica carinata microgreen silver nanoparticles, characterization, safety assessment, and antimicrobial activities

Nanotechnology has been a central focus of scientific investigation over the past decades owing to its versatile applications. The synthesis of silver nanoparticles (AgNPs) through plant secondary metabolites is a cost-effective and eco-friendly approach. The present study employed Brassica carinata microgreen extracts (BCME) to promote the reduction of silver nitrate (AgNO3) salt into Brassica carinata microgreen silver nanoparticles (BCM-AgNPs). The physicochemical properties of the biosynthesized AgNPs were characterized through both spectroscopy and microscopy techniques. Furthermore, the antimicrobial property of the biosynthesized AgNPs was assessed against six selected pathogenic microorganisms, and finally, their safety was evaluated on a normal Vero cell line through an MTT cytotoxicity assay. The UV-visible spectrum revealed that BCM-AgNPs exhibited an absorption peak at 420 nm. The potential functional groups involved in the biosynthesis of AgNPs were identified by Fourier transform infrared (FTIR) analysis. Scanning electron microscopy (SEM) revealed a spherical nature of the biosynthesized AgNPs. Transmission electron microscopy (TEM) analysis revealed the crystallinity of the AgNPs, averaging 34.68 nm in size. X-ray diffraction (XRD) investigation further confirmed the crystalline structure of the AgNPs. The zeta potential exhibited a significant value of − 22.5 ± 1.16 mV. Regarding the antimicrobial potential, BCM-AgNPs exhibited promising antimicrobial activity against the tested pathogens, with a minimum inhibitory concentration (MIC) of 62.5 µg/mL observed in Pseudomonas aeruginosa. Further cytotoxicity assessment of BCM-AgNPs conducted on Vero cells demonstrated their safety. This study presents a novel approach to synthesizing AgNPs using a nutraceutical microgreen, offering a biocompatible and promising alternative for combating multi-drug resistance.

The Remarkable Diversity of Vertebrate Bitter Taste Receptors: Recent Advances in Genomic and Functional Studies.

Bitter taste perception is crucial for animal survival. By detecting potentially harmful substances, such as plant secondary metabolites, as bitter, animals can avoid ingesting toxic compounds. In vertebrates, this function is mediated by taste receptors type 2 (T2Rs), a family of G protein-coupled receptors (GPCRs) expressed on taste buds. Given their vital roles, T2Rs have undergone significant selective pressures throughout vertebrate evolution, leading to frequent gene duplications and deletions, functional changes, and intrapopulation differentiation across various lineages. Recent advancements in genomic and functional research have uncovered the repertoires and functions of bitter taste receptors in a wide range of vertebrate species, shedding light on their evolution in relation to dietary habits and other ecological factors. This review summarizes recent research on bitter taste receptors and explores the mechanisms driving the diversity of these receptors from the perspective of vertebrate ecology and evolution.

Effects of cashew nutshell extract inclusion into a high-grain finishing diet on methane emissions, nutrient digestibility, and ruminal fermentation in beef steers.

By 2050, the U.S. beef industry must produce an extra 40 million tons of beef to satisfy the global demand. Such an increase in inventory will undoubtedly enhance methane (CH4) production from livestock, which should be reduced by over 20%. The addition of plant secondary metabolites, such as anacardic acid present in cashew nutshell extract (CNSE), has shown promising results in reducing CH4 yield, although its effects seemed to be diet dependent. This study evaluated the addition of CNSE to a high-grain diet (85:15 Grain: forage) on in vivo CH4 emissions, nutrients digestibility, performance, feeding behavior, and ruminal fermentation parameters of beef steers. Sixteen Angus crossbred steers [599 ± 40kg of bodyweight (BW)] and six ruminally cannulated crossbred steers (490 ± 51kg of BW) were utilized in a crossover design with 2 experimental periods of 56 d each, composed by 14 d of adaptation, 35 d of measurement, and 7 d of washout. Following adaptation, steers were sorted by BW, and assigned to receive no additive (CON) or CNSE at 5g/steer/d. Data were analyzed using the MIXED procedure of SAS. Inclusion of CNSE increased (P < 0.05) propionate concentration and molar proportion (MP; mol/100mol), tended to decrease acetate MP (P = 0.10), reduced the acetate: propionate (A:P) ratio (P = 0.05), and MP of branched chain volatile fatty acids (P < 0.01). Neither in vitro organic matter digestibility nor in vitro CH4 yield were affected by CNSE inclusion (P > 0.05). Steers receiving CNSE exhibited greater (P < 0.05) final BW, dry matter intake (DMI), and average daily gain (ADG) but lesser (P < 0.05) in vivo CH4 emission rate (g/d), yield (g/kg of DMI), and intensity (g/kg of ADG). Meal length, bunk visit duration, and apparent total tract digestibility of DM increased (P < 0.05) after CNSE addition. Considering CNSE-supplemented steers spent more time in the feedbunk and exhibited higher DMI, CH4 mitigation was unlikely associated with intake reduction. The addition of CNSE to a high-grain diet in beef steers demonstrated significant improvements in animal performance and reduced CH4 emissions, as the result of shifts in ruminal fermentation patterns, favoring propionate instead acetate concentration, leading to a reduction in the A:P ratio. CNSE shows promise as a strategy to enhance beef industry sustainability.