Mechanism study on the improvement of egg white emulsifying characteristic by ultrasound synergized citral: Physicochemical properties, molecular flexibility, protein structure.

As a natural emulsifier, egg white protein (EWP) has great interfacial characteristics and high security, and has broad development prospects. This study explored the impact of ultrasound synergized citral (CI) treatment on the microstructure, molecular flexibility and emulsifying property of EWP, and predicted the interaction between CI and ovalbumin (the main protein in EWP) through molecular docking. The decrease in free amino content and the growth in molecular weight of EWP suggested that CI and proteins were successfully grafted. The results of physicochemical properties revealed that UCEWP (ultrasound synergized citral-treated EWP) had smaller particle size and larger ζ-potential absolute value, which meant that the stability of UCEWP system was enhanced. From the perspective of interfacial characteristics, UCEWP had lower interfacial tension, which remarkably improved its emulsifying property. The emulsifying activity index (EAI) and emulsifying stability index (ESI) of UCEWP were 1.99 times and 3.19 times higher than that of natural EWP (NEWP). Analysis of Fourier transform infrared spectroscopy (FT-IR) and fluorescence spectroscopy illustrated that the secondary and tertiary structures of UCEWP were more disordered and stretched than those of EWPs. Protein microstructure demonstrated that UCEWP presented loose small particle distribution, and correlation analysis reflected that the improvement of molecular flexibility was positively correlated with the enhancement of emulsifying property. These results elucidated that ultrasound synergized CI treatment is an effective mean to improve the molecular flexibility and emulsifying property of EWP, which provides a valuable reference for further application of EWP.

Application of Fe2O3 nanoparticles improves the growth, antioxidant power, flavonoid content, and essential oil yield and composition of Dracocephalum kotschyi Boiss.

Dracocephalum kotschyi Boiss., an endemic and endangered medicinal and aromatic plant in Iran, showcases distinct botanical characteristics and therapeutic promise. According to the IUCN grouping criteria, this plant is facing challenges due to overcollection from its natural habitats. To address this issue, there is an increasing inclination towards cultivating this species within agricultural systems. This study aimed to evaluate the impact of applying Fe2O3 nanoparticles (NPs) at varying concentrations (50, 100, and 200 mg L-1), as well as bulk Fe2O3 at the same concentrations, on the growth, essential oil production, antioxidant capacity, total phenol, and flavonoid content of D. kotschyi. The foliar application of 100 and/or 200 mg L-1 of Fe2O3 NPs resulted in the greatest leaf length and dry weight, while Fe2O3 NPs at the level of 100 mg L-1 led to the highest leaf/stem ratio. Additionally, spraying 200 mg L-1 of Fe2O3 NPs and all concentrations of bulk Fe2O3 positively impacted chlorophyll and carotenoid levels. Both nano and bulk Fe2O3 supplements stimulated H2O2 production and subsequently enhanced enzymatic antioxidant activity. The use of 50 mg L-1 of Fe2O3 NPs resulted in the highest flavonoid content and non-enzymatic antioxidant activity. Meanwhile, the highest essential oil content and yield was achieved by the application of 50 and/or 100 mg L-1 Fe2O3 NPs. The addition of low concentration of Fe2O3 NPs (50 mg L-1) resulted in a significant increase in the concentration of geranial, while a higher supply of Fe2O3 NPs (200 mg L-1) significantly decreased the percentage of neral in the essential oil. Overall, the application of Fe2O3 NPs demonstrated significant potential for increased biomass, enhanced yield, essential oil production, and phytochemical attributes. The findings highlight the versatility of Fe2O3 NPs at optimal concentrations, acting as both a nano-fertilizer and a nano-inducer, promoting the production and accumulation of valuable secondary metabolites in plants.

Comparative Study of the Protective Effects of Citral, Thymoquinone, and Silymarin on Methotrexate-induced Cardiotoxicity in Rats.

Objectives: Methotrexate (MTX), an immunosuppressant and anti-cancer medication, can harm the heart. The goal of the current investigation was to assess the cardiotoxicity caused by MTX and the potential cardioprotective properties of silymarin, citral, and thymoquinone as antioxidants. Methods: Forty-eight rats were divided into six groups, which included control, MTX, cosolvent, citral, thymoquinone, and silymarin groups. At the end of the study, the rats were anesthetized (ketamine and xylazine) and killed using CO2. Their blood samples were collected to measure the enzymatic activities of creatine kinase-myoglobin binding (CK-MB), creatine phosphokinase (CPK), and lactate dehydrogenase (LDH). Also, the heart tissue was sampled to determine the antioxidant capacity and examine the histopathology. Results: The findings revealed that the activity of CPK, CK-MB, and LDH enzymes significantly reduced in the thymoquinone treatment group compared to the MTX group (p < 0.05). On the other hand, total antioxidant capacity was significantly increased in the thymoquinone group compared to the MTX group (p < 0.05). The pathological modifications (i.e. severe congestion, edema fluid, the presence of inflammatory cells around the blood vessels, mild to moderate hemorrhaging between cardiac muscle fibers) were seen in the MTX group. The treatment groups, particularly thymoquinone, did not experience any appreciable pathological changes. Conclusion: The thymoquinone was found to have the strongest protective effect against the heart damage caused by MTX.

Phylogenetically distant enzymes localized in cytosol and plastids drive citral biosynthesis in lemongrass.

Citral, a naturally occurring acyclic monoterpene aldehyde, is present in the essential oils of various plants, but only a few produce it in abundance. Despite its importance as a key aroma molecule, knowledge regarding the in-planta biosynthesis of citral and its metabolic origin remains limited. Here, we have elucidated the functions of an alcohol dehydrogenase (CfADH1) and an aldoketo-reductase (CfAKR2b) in citral biosynthesis in lemongrass (Cymbopogon flexuosus), one of the most cultivated aromatic crops for its citral-rich essential oil. Expression of both CfADH1 and CfAKR2b showed correlation with citral accumulation in different developmental stages. Recombinant CfADH1 and CfAKR2b, despite their sequence unrelatedness, catalyzed citral formation from geraniol with NADP cofactor. Virus-induced gene silencing in lemongrass and transient expression in lemon balm (Melissa officinalis) demonstrated the in-planta involvement of CfADH1 and CfAKR2b in citral biosynthesis. While CfADH1 exhibited a dual cytosolic/plastidial localization, CfAKR2b was localized to the cytosol. This was supported by higher citral-forming activity in the cytosolic fraction than in the chloroplast fraction of lemongrass leaf extract. Moreover, feeding lemongrass seedlings with inhibitors specific to the cytosolic mevalonate pathway and the plastidial methylerythritol phosphate pathway, combined with volatile profiling, supported the involvement of both pathways in citral formation. Taken together, our results indicate that high citral production has evolved in lemongrass through the recruitment of phylogenetically distant enzymes localized in both the cytosol and plastids.

Exploring the physiological response differences of ß-caryophyllene, linalool and citral inhalation and their anxiolytic potential.

Essential oils with ß-caryophyllene, citral, and linalool as key compounds often exhibit some anti-anxiety like effects in aromatherapy. However, evidence of the effect of these three compounds through human inhalation remains limited. It is worth exploring their potential anxiolytic effect through the olfactory pathway, and finding out whether the three compounds lead to different physiological responses. A total of 48 subjects were randomly assigned to three odor (ß-caryophyllene, citral, and linalool) inhalation groups and one control (odorless jojoba oil) group. Stress stimulation was induced using n-back and mental arithmetic tasks. The odor was administered before the task test session. Assessments including the State-Trait Anxiety Inventory (STAI), electroencephalogram (EEG) activities, facial expressions, several physiological indicators, and a self-report scale of subjective perception of the odor environments were carried out. The changes before and after inhalation, as well as the inter-group differences, were analyzed. Both ß-caryophyllene and citral inhalation led to a significant decrease in anxiety levels, while only ß-caryophyllene resulted in a notable reduction across both sub-scales of STAI. Following the odor inhalation, heart rate significantly decreased in all three groups, with the ß-caryophyllene group exhibiting the most pronounced decline. While the systolic blood pressure of the linalool group demonstrated a statistically significant difference. Regarding facial expressions, ß-caryophyllene significantly increased the ratio of 'Happiness' and decreased the ratio of 'Fear'. In the non-task state, citral reduced the power of frontal alpha, delta, and theta waves while ß-caryophyllene had a similar effect. All odor inhalation groups showed increased delta and theta waves after the task compared with the control group, with the ß-caryophyllene group having notably lower frontal beta waves. ß-Caryophyllene and citral exhibited good anti-anxiety effects. Subjects receiving different odors showed different EEG and physiological responses, indicating the differences in emotional regulation ways among the three compounds.

Influence of Essential Oil Composition on Antioxidant and Antibacterial Activities of Three Cultivars of Cymbopogon Flexuosus: In Vitro and In Silico Study.

The present study investigates and compares the chemical composition, antioxidant, and antibacterial properties of lemongrass essential oils (LEOs) extracted from fresh leaves of three cultivars of C. flexuosus: Krishna (CF-KA), Cauvery (CF-CA), and Nima (CF-NI), grown in Chhattisgarh plains. Analysis through gas chromatography techniques revealed that citral content was highest in CF-NI (79.82±1.00 %), followed by CF-KA (69.75±2.70 %) and CF-CA (54.75±1.22 %). In vitro antioxidant experiments demonstrated that CF-CA had better scavenging capacity in DPPH (SC50=164.55±9.35 µg/mL) and ABTS (SC50=4.76±0.57 GEAC/g) free radical scavenging assays. The in vitro antibacterial experiments against Staphylococcus aureus (MTCC3160) and Escherichia coli (MTCC1687) demonstrated CF-NI's enhanced antibacterial efficacy with significant inhibition zones and low MIC values. In silico molecular docking results revealed that LEO compounds like caryophyllene oxide, humulene epoxide, ß-caryophyllene etc. have better binding affinities towards targeted protein molecules responsible for bacterial cell mechanisms and production of reactive oxygen species (ROS) compared to their native ligands. Variations in biological activities among cultivars were potentially linked to the proportion of phytoconstituents in their chemical composition.

Evaluation of antifungal and apoptotic effects of linalool, citral, and carvacrol separately and in combination with nystatin against clinical isolates of Pichia kudriavzevii.

Pichia kudriavzevii (formerly Candida krusei) poses a significant threat to immunocompromised patients due to its inherent resistance to various antifungal drugs. This study explored the anticandidal potential of citral, linalool, and carvacrol in combination with nystatin against P. kudriavzevii strains.Using the microdilution method following CLSI guidelines, Minimum Inhibitory Concentrations (MICs) and fungicidal concentrations (MFCs) were determined. Citral exhibited MIC values ranging from 50 to 100 µg/ml, averaging 70.24 ± 16.99 µg/ml, while carvacrol had MIC values of 50 to 100 µg/ml, averaging 86.90 ± 16.99 µg/ml. Linalool demonstrated weaker antifungal activity, with MIC values between 100 and 200 µg/ml, averaging 150 ± 38.73 µg/ml. The study assessed the synergistic effectsof these phenols with nystatin through fractional inhibitory concentration indices (FICIS). In addition, flow cytometry was employed to assess apoptosis induction in P. kudriavzevii cells.Carvacrol displayed a remarkable synergistic effect in combination with nystatin against all 21 isolates tested. Conversely, linalool showed synergy in 17 isolates, while citral exhibited synergy in only 2 isolates. These findings highlight distinct patterns of synergy between the different compounds and nystatin against P. kudriavzevii. Also, Carvacrol emerged as the most potent inducer of apoptosis across all P. kudriavzevii strains, followed by citral and linalool. This suggests that carvacrol not only possesses a stronger antifungal effect but also has a more pronounced ability to trigger programmed cell death in P. kudriavzevii. In conclusion, the study supports the potential of carvacrol, citral and linalool, as anticandidal agents, suggesting their supplementation with nystatin for treating P. kudriavzevii infections.

Citral: A potent inhibitor of advanced glycation end products.

Advanced glycation end products (AGEs), which are produced during food processing, pose health risks to humans. This study found that citral (Cit) effectively inhibited the formation of both fluorescent and non-fluorescent AGEs in the bovine serum albumin (BSA)-glucose (Glc) system. Cit achieved an average inhibition rate of over 80 % for fluorescent AGEs and reduced the levels of N-ε-carboxymethyllysine (CML) and N-ε-carboxyethyllysine (CEL) by up to 45.85 % and 59.32 %, respectively. The comprehensive characterizations and high-resolution mass spectrometry analysis demonstrated that the carbonyl group and CC group present on Cit could compete with Glc for the amino groups on BSA, thereby reducing the formation of AGEs. Additionally, the cytotoxicity assay demonstrated that the BSA-Cit adducts were non-toxic. This research indicated that Cit was a potent and safe inhibitor of AGEs.

Characterization and antimicrobial activity of essential oils extracted from lemongrass (Cymbopogon flexuosus) using microwave-assisted hydro distillation.

Lemongrass (Cymbopogon flexuosus) essential oil (LGEO) contains α-citral, ß-citral and other phytochemicals extracted using various methods. This research extracted essential oils using steam distillation (SD) and microwave-assisted hydro distillation (MAHD) to maximize quantity and purity. LGEO was tested for antibacterial properties. LGEO was extracted using SD and compared to MAHD output based on oil production and chemical composition. We performed GCMS to characterize LGEO. Fourier transform infrared spectroscopy (FTIR) used for quantum chemical analysis. Spectroscopic analysis showed that SD extracted secondary metabolites (ethyl-linalool, isogeranial, ß-citral, α-citral, geranyl acetate, and caryophyllene) yielded 9.7 %, 11.5 %, 35.4 %, 13.4 %, 6.4 %, and 6.4 %, respectively, while MAHD yielded 10.2 %, 13.4 %, 43.2 %, 17.3 %, 6.9 %, and 7.3 %. MAHD extracted α and ß citral content was better than SD extraction technique. FTIR spectroscopy and quantum chemistry analysis showed extracted oil chemical composition, electronic structure of α and ß citral isomers. In the disc-diffusion experiment, both extracts were effective against Gram-positive and Gram-negative bacteria and harmful fungi. LGEO from SD and MAHD extraction (30 mg/mL) demonstrated disc diffusion assay antibacterial efficacy against microorganisms. The two extracts effectively inhibited microorganisms with MIC values of 3.75 and 7.5 µg/mL. It can be concluded that, LGEO have greater antimicrobial activity in MAHD extraction.

Inhibitory activity and antioomycete mechanism of citral against Phytophthora capsici.

The natural terpenoid citral has antifungal activity against multiple fungi, but its bioactivity against oomycetes is unclear. Therefore, this study investigated the antioomycete activity and mechanism of citral against Phytophthora capsici, a highly destructive invasive oomycete. Results showed that citral not only had a great inhibition on the mycelial growth of P. capsici (EC50 = 94.15 mg/L), but also had a significant inhibition on multiple spores, such as sporangia formation, zoospore discharge and zoospore germination. Citral at 4000 mg/L exhibited favorable protective (73.33%) and curative efficacy (55.11%) against pepper Phytophthora blight. Citral significantly damaged the hyphal morphology, disrupted the cell membrane integrity, increased the permeability of cell membrane, and increased the glycerol content in P. capsici. A total of 250 upregulated and 288 downregulated proteins were identified in iTRAQ-based quantitative proteomic analysis. Downregulated proteins were mostly enriched in pathways of ABC transporters, cyanoamino acid metabolism and starch and sucrose metabolism, suggesting an inhibition of citral on transmembrane transporter (e.g., ABC transporters) and pathogenicity (e.g., ß-glucosidases) proteins. Upregulated proteins were enriched in biosynthesis of unsaturated fatty acids, pyruvate metabolism and glycolysis/gluconeogenesis, suggesting an activation of citral on energy generation proteins, including acyl-CoA oxidase, D-lactate dehydrogenase, pyruvate kinase, acetyl-CoA synthetase and phosphoenolpyruvate carboxykinase. Biochemical and iTRAQ analysis suggested that cell membrane may be the target of citral in P. capsici.

The aroma transformation of Japanese sea bass (Lateolabrax japonicas) through endogenous enzyme incubation during the lag phase of attached microorganisms.

Endogenous enzymes play a crucial role in determining fish product aroma. However, the attached microorganisms can promote enzyme production, making it challenging to identify specific aromatic compounds resulting from endogenous enzymes. Thus, we investigated the aroma transformation of Japanese sea bass through enzymatic incubation by controlling attached microorganisms during the lag phase. Our results demonstrate that enzymatic incubation significantly enhances grassy and sweet notes while reducing fishy odors. These changes in aroma are associated with increased levels of 10 volatile compounds and decreased levels of 3 volatile compounds. Among them, previous studies have reported enzyme reaction pathways for octanal, 1-nonanal, vanillin, indole, linalool, geraniol, citral, and 6-methyl-5-hepten-2-one; however, the enzymatic reaction pathways for germacrene D, beta-caryophyllene, pristane, 1-tetradecene and trans-beta-ocimene remain unclear. These findings provide novel insights for further study to elucidate the impact of endogenous enzymes on fish product aromas.

Novel pesticides design: co-encapsulation of citral and cyproconazole for the control of Botrytis cinerea using biocompatible nano-carriers.

BACKGROUND: Growing concerns about sustainability have driven the search for eco-friendly pest management solutions. Combining natural and synthetic compounds within controlled release systems is a promising strategy. This study investigated the co-encapsulation of the natural compound citral (Cit) and the synthetic antifungal cyproconazole (CPZ) using two water-based nanocarriers: solid lipid nanoparticles (SLNs) and chitosan nanoparticles (CSNPs). RESULTS: Both CSNPs and SLNs loaded with Cit + CPZ displayed superior antifungal activity against Botrytis cinerea compared to free compounds. Notably, CSNPs with a 2:1 Cit:CPZ ratio exhibited the highest efficacy, achieving a minimum inhibitory concentration (MIC100) of < 1.56 µg mL-1, lower than the 12.5 µg mL-1 of non-encapsulated compounds. This formulation significantly reduced the required synthetic CPZ while maintaining efficacy, highlighting its potential for environmentally friendly pest control. CONCLUSION: The successful co-encapsulation of Cit + CPZ within CSNPs, particularly at a 2:1 ratio, demonstrates a promising approach for developing effective and sustainable antifungal formulations against B. cinerea. © 2024 Society of Chemical Industry.

Cymbopogon citratus Allelochemical Volatiles as Potential Biopesticides against the Pinewood Nematode.

Traditional pesticides are based on toxic compounds that can reduce biodiversity, degrade the environment, and contribute to less healthy living. Plant allelochemicals can provide more environmentally friendly and sustainable alternatives. Essential oils (EOs) are complex mixtures of plant secondary metabolites that show strong biological activities. In the present study, the EOs of Cymbopogon citratus were screened for activity against the pinewood nematode (PWN), the causal agent of pine wilt disease. To understand their nematicidal properties, EOs were fractioned into hydrocarbon molecules and oxygen-containing compounds, and their main compounds were acquired and tested separately against the PWN. The EO oxygen-containing molecules fraction was highly active against the PWN (EC50 = 0.279 µL/mL), with citral and geraniol showing higher activities (EC50 = 0.266 and 0.341 µL/mL, respectively) than emamectin benzoate (EC50 = 0.364 µL/mL), a traditional nematicide used against the PWN. These compounds were additionally reported to be less toxic to non-target organisms (fish, invertebrates, and algae) and safer to human health (with higher reported toxicity thresholds) and predicted to exert fewer environmental impacts than traditional nematicides. Resorting to approved natural compounds can quickly leverage the development of sustainable alternatives to traditional nematicides.

Larvicidal activity of ß-Citral: An In-vitro and In-silico study to understand its potential against mosquito.

Tropical and subtropical regions face millions of deaths from mosquito-borne illnesses yearly. Insecticides prevent transmission but pose health risks like dermatitis and allergies. The primary objective was to mitigate the recurring dependence on synthetic insecticides, thereby curbing the development of mosquito resistance. Leaves of Cymbopogon flexuosus (lemongrass) was collected from Mayurbhanj, India, processed, then extracted by steam distillation for essential oils & analyzed spectroscopically. Larvicidal assays were performed across varying concentrations, revealing the significant mortality induced by the Cymbopogon flexuosus extract against Anopheles stephensi larvae. 3D structure was modelled by using G protein-coupled receptors (GPCR) sequence and structural stability was also validated. After docking the binding free energy was determined from GPCR protein with ß-citral complex. Molecular dynamics (MD) study was conducted on the docked pose that displayed an optimal interactome profile. The larvicidal assay at the 12th and 24th hour revealed the highest LC50 (lethal concentration) of 23.493 ppm and 19.664 ppm . ß-Citral has a high binding affinity and an identifiable binding site, which suggests that it may play a larvicidal role in regulating the receptor's function by creating stable complexes with it. ß-Citral from lemongrass oils has potential larvicidal activity and effective against GPCR family 1 of mosquito and highly effective repellents against mosquito-borne diseases.

Enhancing Selectivity and Inhibitory Effects of Chemotherapy Drugs Against Myelogenous Leukemia Cells with Lippia alba Essential Oil Enriched in Citral.

Acute myelogenous leukemia (AML) is one of the most lethal cancers, lacking a definitive curative therapy due to essential constraints related to the toxicity and efficacy of conventional treatments. This study explores the co-adjuvant potential of Lippia alba essential oils (EO) for enhancing the effectiveness and selectivity of two chemotherapy agents (cytarabine and clofarabine) against AML cells. EO derived from L. alba citral chemotype were produced using optimized and standardized environmental and extraction protocols. Rational fractionation techniques were employed to yield bioactive terpene-enriched fractions, guided by relative chemical composition and cytotoxic analysis. Pharmacological interactions were established between these fractions and cytarabine and clofarabine. The study comprehensively evaluated the cytotoxic, genotoxic, oxidative stress, and cell death phenotypes induced by therapies across AML (DA-3ER/GM/EVI1+) cells. The fraction rich in citral (F2) exhibited synergistic pharmacological interactions with the studied chemotherapies, intensifying their selective cytotoxic, genotoxic, and pro-oxidant effects. This shift favored transitioning from necrosis to a programmed cell death phenotype (apoptotic). The F2-clofarabine combination demonstrated remarkable synergistic anti-leukemic performance while preserving cell integrity in healthy cells. The observed selective antiproliferative effects may be attributed to the potential dual prooxidant/antioxidant behavior of citral in L. alba EO.

Insights into chemistry, extraction and industrial application of lemon grass essential oil -A review of recent advances.

Lemongrass essential oil (LEO), extracted from high-oil lemongrass, gains prominence as a versatile natural product due to growing demand for safe health solutions. LEO comprises beneficial compounds like citral, isoneral, geraniol, and citronellal, offering diverse pharmacological benefits such as antioxidant, antifungal, antibacterial, antiviral, and anticancer effects. LEO finds applications in food preservation, cosmetics, and pharmaceuticals, enhancing profitability across these sectors. The review focuses on the extraction of LEO, emphasizing the need for cost-effective methods. Ultrasound and supercritical fluid extraction are effective in reducing extraction time, increasing yields, and enhancing oil quality. LEO shows promise as a valuable natural resource across industries, with applications in packaging, coating, and film development. LEO's ability to extend the shelf life of food items and impart natural flavors positions it as a valuable asset. Overall, the review emphasizes LEO's therapeutic, antimicrobial, and antioxidant properties, strengthening its potential in the food, pharmaceutical, and cosmetic sectors.

Comparative investigation on the aroma profiles of edible citrus flowers in the main organs and different developmental stages.

Pomelo flowers emit a strong fragrance and give aromatic odors. Volatile compounds from pomelo flowers were analyzed at three developmental stages and in the main organs by molecular sensory science. A total of 134 volatiles including 25 odorants, were analyzed by gas chromatography-mass spectrometry/pulsed flame photometric detector (GC-MS/PFPD) and multidimensional GC-MS/olfactory (MDGC-MS/O). The total volatile content varied among pomelo flowers at different developmental stages (stage-III > stage-II > stage-I) and among different organs of pomelo flowers (petal > pistil > stamen). Linalool was an important odorant with a high OAV, and floral/fruity comprised the predominant aroma profile. Four odorants, ethyl 2-methylbutanoate, linalool, ß-myrcene, and 2-butenal, were selected based on variable importance in projection (VIP) values and contributed mainly to the discrimination of pomelo flowers at three different developmental stages. Linalool, ß-myrcene, d-limonene, and ethyl hexanoate were potential markers for evaluating flavor differences in pomelo floral organs.

Identification and functional analysis of a deduced geraniol synthase from Camphora officinarum.

The market demand for essential oil containing citral is increasing. Our research group identified a rare chemotype of Camphora officinarum whose leaves are high in citral content by examining over 1000 wild trees across the entire native distribution area of C. officinarum in China. Because C. officinarum is suitable for large-scale cultivation, it is therefore seen as a promising source of natural citral. However, the molecular mechanism of citral biosynthesis in C. officinarum is poorly understood. In this study, transcriptomic analyses of C. officinarum with different citral contents revealed a strong positive correlation between the expression of a putative geraniol synthase gene (CoGES) and citral content. The CoGES cDNA was cloned, and the CoGES protein shared high similarity with other monoterpene synthases. Enzymatic assays of CoGES with geranyl diphosphate (GPP) as substrate yielded geraniol as the single product, which is the precursor of citral. Further transient expression of CoGES in Nicotiana benthamiana resulted in a higher relative content of geranial and the appearance of a new substance, neral. These findings indicate that CoGES is a geraniol synthase-encoding gene, and the encoded protein can catalyze the transformation of GPP into geraniol, which is further converted into geranial and neral through an unknown mechanism in vivo. These findings expand our understanding of citral biosynthesis in Lauraceae plants. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-024-01463-4.

Inhibition and Mechanism of Citral on Bacillus cereus Vegetative Cells, Spores, and Biofilms.

Bacillus cereus causes food poisoning by producing toxins that cause diarrhea and vomiting and, in severe cases, endocarditis, meningitis, and other diseases. It also tends to form biofilms and spores that lead to contamination of the food production environment. Citral is a potent natural antibacterial agent, but its antibacterial activity against B. cereus has not been extensively studied. In this study, we first determined the minimum inhibitory concentrations and minimum bactericidal concentrations, growth curves, killing effect in different media, membrane potential, intracellular adenosine triphosphate (ATP), reactive oxygen species levels, and morphology of vegetative cells, followed by germination rate, morphology, germination state of spores, and finally biofilm clearance effect. The results showed that the minimum inhibitory concentrations and minimum bactericidal concentrations of citral against bacteria ranged from 100 to 800 µg/mL. The lag phase of bacteria was effectively prolonged by citral, and the growth rate of bacteria was slowed down. Bacteria in Luria-Bertani broth were reduced to below the detection limit by citral at 800 µg/mL within 0.5 h. Bacteria in rice were reduced to 3 log CFU/g by citral at 4000 µg/mL within 0.5 h. After treatment with citral, intracellular ATP concentration was reduced, membrane potential was altered, intracellular reactive oxygen species concentration was increased, and normal cell morphology was altered. After treatment with citral at 400 µg/mL, spore germination rate was reduced to 16.71%, spore morphology was affected, and spore germination state was altered. It also had a good effect on biofilm removal. The present study showed that citral had good bacteriostatic activity against B. cereus vegetative cells and its spores and also had a good clearance effect on its biofilm. Citral has the potential to be used as a bacteriostatic substance for the control of B. cereus in food industry production.

Enhancement of lemongrass essential oil physicochemical properties and antibacterial activity by encapsulation in zein-caseinate nanocomposite.

Essential oils (EOs) represent a pivotal source for developing potent antimicrobial drugs. However, EOs have seldom found their way to the pharmaceutical market due to their instability and low bioavailability. Nanoencapsulation is an auspicious strategy that may circumvent these limitations. In the current study, lemongrass essential oil (LGO) was encapsulated in zein-sodium caseinate nanoparticles (Z-NaCAS NPs). The fabricated nanocomposite was characterized using dynamic light scattering, Fourier-transform infrared spectroscopy, differential scanning calorimetry, and transmission electron microscopy. The antimicrobial activity of LGO loaded NPs was assessed in comparison to free LGO against Staphylococcus epidermidis, Enterococcus faecalis, Escherichia coli, and Klebsiella pneumoniae. Furthermore, their antibacterial mechanism was examined by alkaline phosphatase, lactate dehydrogenase, bacterial DNA and protein assays, and scanning electron microscopy. Results confirmed the successful encapsulation of LGO with particle size of 243 nm, zeta potential of - 32 mV, and encapsulation efficiency of 84.7%. Additionally, the encapsulated LGO showed an enhanced thermal stability and a sustained release pattern. Furthermore, LGO loaded NPs exhibited substantial antibacterial activity, with a significant 2 to 4 fold increase in cell wall permeability and intracellular enzymes leakage versus free LGO. Accordingly, nanoencapsulation in Z-NaCAS NPs improved LGO physicochemical and antimicrobial properties, expanding their scope of pharmaceutical applications.