Search Results (69)

Plant callus cultures are a sustainable alternative for producing bioactive secondary metabolites, but their low yields limit industrial applications. Carob (Ceratonia siliqua L.) is rich in medicinally valuable compounds, yet conventional cultivation faces challenges. To address this, we use biofortified calcium phosphate nanoparticles, which refer to CaP-NPs that have been enriched with bioactive compounds via green synthesis using Jania rubens extract, thereby enhancing their functional properties as elicitors in carob callus. CaP-NPs were green-synthesized using Jania rubens extract and applied to 7-week-old callus cultures at 0, 25, 50, and 75 mg/L concentrations. At the optimal concentration (50 mg/L), CaP-NPs increased callus fresh weight by 23.9% and dry weight by 35.1%. At 50 mg/L CaP-NPs, phenolic content increased by 95.7%, flavonoids by 34.4%, tannins by 131.8%, and terpenoids by 211.9% compared to controls. Total antioxidant capacity rose by 76.2%, while oxidative stress markers malondialdehyde (MDA) and hydrogen peroxide (H₂O₂) decreased by 34.8% and 14.1%, respectively. Gene expression analysis revealed upregulation of PAL (4-fold), CHI (3.15-fold), FLS (1.16-fold), MVK (8.3-fold), and TA (3.24-fold) at 50 mg/L CaP-NPs. Higher doses (75 mg/L) induced oxidative damage, demonstrating a hormetic threshold. These findings indicate that CaP-NPs effectively enhance secondary metabolite production in carob callus by modulating biosynthetic pathways and redox balance, offering a scalable, eco-friendly approach for pharmaceutical and nutraceutical applications. Full article

Background: Carob (Ceratonia siliqua L.) pulp flour is primarily used in the food industry. As a rich source of bioactive compounds, particularly polyphenols, it holds promise for pharmaceutical formulation research and development. Objectives: This study focused on developing liposomal particles loaded with carob pulp extract using the proliposome method, followed by modifications through UV irradiation and sonication. Methods: The resulting liposomes were analyzed for encapsulation efficiency, vesicle size, polydispersity index (PDI), mobility, zeta potential, viscosity, surface tension, density, antioxidant activity, FT-IR spectra, and release kinetics under simulated gastrointestinal conditions. In addition, nanoparticle tracking analysis and transmission electron microscopy (TEM) were used for liposomal characterization. Results: The findings revealed a high encapsulation efficiency across all samples (>70%). The particle size and PDI measurements confirmed the presence of a multilamellar and uniform liposomal system before post-processing modifications. The medium value of zeta potential suggested a moderately electrostatically stabilized liposomal suspension. The sonicated liposomes demonstrated a higher concentration of vesicles in comparison to non-treated and UV-irradiated samples. TEM analysis revealed purified liposomal vesicles with preserved structural integrity. Encapsulation, as well as UV irradiation and sonication of liposomes, did not diminish the extract’s anti-DPPH activity. However, the ABTS radical scavenging potential of the pure extract was significantly lower compared to its encapsulated counterparts. UV irradiation and sonication notably reduced the anti-ABTS capacity of the extract-liposome system. Monitoring the release of bioactive compounds demonstrated controlled delivery from liposomal particles under simulated gastrointestinal conditions. Conclusions: Overall, liposomal formulations of carob pulp extract exhibit significant potential for further development as a functional food ingredient or for use in the prevention and treatment of various diseases. Full article

Glycation in biological systems contributes to the development of chronic diseases, particularly under conditions of hyperglycemia and oxidative stress. This study evaluated the antiglycation and methylglyoxal (MGO)-trapping capacities of aqueous and methanolic extracts of carob flour. The methanolic extract exhibited significantly higher bioactive compounds, containing 1.4-fold more total phenolics and 1.6-fold more flavonoids than the aqueous extract, as well as 1.2- and 1.8-fold-higher antioxidant activity. Antiglycation activity was assessed using bovine serum albumin (BSA)–glucose and BSA–MGO in vitro models, where the methanolic extract consistently outperformed the aqueous extract. At 25 mg/mL, the formation of advanced glycation end-products was inhibited by 81.0% in the BSA–glucose model and nearly 70% in the BSA–MGO model. These findings were supported by lower IC₅₀ values for the methanolic extract (6.6 vs. 10.8 mg/mL and 9.4 vs. 16.6 mg/mL). MGO-trapping capacity was also higher for the methanolic extract, reaching 97% with 25 mg/mL after 168 h. The superior antiglycation and MGO-trapping activities of the methanolic extract are attributed to its higher content of gallic acid and other phenolic compounds with known bioactivities. These results highlight the potential of carob-based formulations as functional ingredients with preventive applications against glycation-associated pathologies. Full article

Background: Agro-industrial by-products, such as carob and brewer’s spent grain (BSG), have gained increasing attention as sustainable feed ingredients capable of enhancing the nutritional and functional value of Tenebrio molitor larvae. This study aimed to evaluate the effects of carob and BSG supplementation of growth substrates on larval performance and the functional properties of the resulting insect meals. Methods: Seven-week-old larvae were reared for 14 days on the following three diets: control (wheat bran), wheat bran + carob (Trt1), and wheat bran + BSG (Trt2). Larval weight, substrate consumption, and survival were recorded. After processing, insect meals were analyzed for chemical composition, total phenolic content (TPC), and antioxidant activity in both water- and lipid-soluble extracted fractions. Results: Both treatment groups showed significantly higher final weights than the control (Ctrl: 104.9 ± 2.69 g; Trt1: 114.8 ± 1.26 g; Trt2: 116.9 ± 1.07 g; p < 0.05). TPC was highest in the control for both fractions, and the antioxidant activity was similar between the Ctrl and Trt1 groups in the water-soluble fraction but significantly lower in the Trt2 group (p < 0.05). Conclusions: These findings highlight the potential of carob and BSG for sustainable insect farming, though their functional benefits may depend on the inclusion levels and their interaction with insect metabolism. Full article

The integration of functional ingredients into staple foods like bread offers a promising strategy for improving public health. Carob (Ceratonia siliqua L.) flour, rich in bioactive compounds, has potential as a functional additive. However, its incorporation into bread negatively affects dough behavior and product quality due to high levels of insoluble dietary fibers. This study investigates the use of carob extract (PCE) as a functional additive to enhance the nutritional and bioactive profile of bread while preserving its rheological behavior and sensory quality. PCE was obtained via microwave-assisted extraction and spray drying, and incorporated into bread formulations at 1%, 3%, and 5%. The addition of PCE reduced water absorption by 1.5% and increased dough stability three times. Dough resistance increased by 15%, while extensibility decreased by 5%. The viscoelastic properties of dough were preserved, as the storage modulus increased and Tan δ values remained stable. Changes in specific volume, crumb texture, crumb porosity, and bread color of produced bread with PCE were minimal; however, aroma, taste, and overall sensory quality were improved. Additionally, the incorporation of PCE resulted in a significant increase in total phenolic content and antioxidant activity, indicating an enhancement of the bread’s functional properties. These improvements were achieved without negatively affecting the dough rheology or bread quality parameters. Overall, the findings suggest that PCE can be a promising functional ingredient in bread formulations, contributing to both nutritional value and technological performance. Full article

Carob fruit utilization remains limited, with most of their commercial value derived from locust bean gum, which is obtained from seed endosperm. Efficient extraction requires dehusking, which is traditionally performed under harsh conditions. This study aims to develop and optimize a milder, more sustainable dehusking method while preserving seed quality for industrial applications. Various aqueous-based solvents were tested, leading to the selection of metanesulfonic acid (CH₄O₃S). A Box–Behnken design with response surface methodology optimized the process, using husk removal efficiency as the response variable. The optimized conditions were 24.5 g of seeds treated in 50 mL of a solvent mixture (41% CH₄O₃S and 59% H₂SO₄) at 90 °C for 10 min, followed by washing by 5 min with water (87 mL). The treated seeds were analyzed using colorimetry assays and diffusive reflectance spectroscopy and benchmarked against both pristine and dehusked seeds from a local company. The resulting seeds remained morphologically intact and exhibited appealing color characteristics comparable to commercial samples. The optimized method ensured intact seed morphology and color characteristics comparable to commercial standards, offering a viable alternative to conventional H₂SO₄-based dehusking. Furthermore, this study also highlights for the first time the effectiveness of diffusive reflectance spectroscopy as a rapid and straightforward tool for assessing the dehusking process. Full article

The carob tree (Ceratonia siliqua L.) is a defining species of the Mediterranean region, and its fruit, the carob pod, has seen a notable increase in economic interest in recent years, primarily due to the production of locust bean gum (E410), a widely used food additive derived from the seeds. The remainder of the fruit, the carob pulp, comprises 80–90% of the fruit’s weight and is typically considered a by-product, with its primary application being in animal feed. This study focused on obtaining cellulose-rich extracts from selected carob varieties cultivated in the Mediterranean region. A comprehensive physicochemical characterization of these cellulose-rich fractions was conducted, including the assessment of their antioxidant properties, specifically total phenolics and antioxidant capacity measured by the FRAP, ABTS, and CUPRAC methods. The findings reveal that carob pulp is an excellent source of carbohydrates, including soluble sugars, which constitute 33–45% of the pulp’s fresh weight, depending on the variety, and cell wall polysaccharides. The cell wall polymers, with cellulose as the predominant component, account for approximately 45% of the fresh pulp weight. Notable amounts of other polysaccharides, such as pectins and hemicelluloses, were also identified. Among the studied varieties, Bugadera and Rotjal stood out as exceptional sources of cellulose-rich extracts. Carob pulp was also found to be rich in antioxidant compounds, reflected in its high antioxidant capacity. In particular, the Bugadera variety, grown under irrigated conditions, exhibited a significant concentration of phenolic compounds (24.4 mg gallic acid equivalents per gram of pulp) and high antioxidant activity across all methods used, with ABTS measurements reaching up to 391.5 mg Trolox equivalents per gram of pulp. In conclusion, these results underscore the significant potential of carob pulp as a source of valuable cellulose-rich extracts, offering applications beyond its traditional use as animal feed. By exploring these new possibilities, the economic and environmental sustainability of carob cultivation could be greatly enhanced, contributing to the broader valorization of this iconic Mediterranean fruit. Full article

Ceratonia siliqua L. (Fabaceae) is an evergreen sclerophyllous species that successfully overcomes the challenges of the Mediterranean climate. Commonly, biosynthesis of secondary metabolites is a major reaction of the plants thriving in the Mediterranean formations against temperature stress. Due to concerns about the climate crisis, we studied the impact of 6-day low (5 °C) and high (40 °C) temperature stress on young carob seedlings. In stressed plants, mainly the heat-treated, the leaves appear xeromorphic. Parameters of the physiology of the plants such as chlorophyll-a and -b, total phenolic content, and oxidative stress were measured and presented via Principal Component Analysis. Chlorophyll-a and -b contents are inferior in cold-stressed leaves while heat-stressed leaves accumulate more phenolics and experience higher oxidative stress as compared to their cold-stressed counterparts. The phytochemical profile of different extracts obtained from stressed carob leaves was identified so as to gain insight into metabolites produced under stress. Moreover, LC-HRMS/MS metabolomic workflow was utilized for the discovery of biomarkers, over- or under-regulated in stressed conditions. The antimicrobial activity of carob leaf extract fractions was assessed against six human pathogen strains and three phytopathogen bacterial strains. MeOH-H₂O and dichloromethane (DCM) extracts presented notable activity against Candida albicans and Saccharomyces cerevisiae, while DCM extracts inhibited the growth of Erwinia amylovora. We may conclude that carob tree exposure to temperature stress does not have a significant influence on secondary metabolic pathways. Full article

Background/Objectives: Hyperlipidemia is a serious risk factor for cardiovascular diseases and liver steatosis. In this work, we explored the effect of an herbal formula (CBF) containing immature Ceratonia siliqua pods and Ocimum basilicum extracts on lipid metabolism disorders and lipoprotein-rich plasma (LRP) oxidation in mice. Methods: The phenolic composition was determined using HPLC-DAD analysis. The antioxidant activity was studied using various in vitro methods. Acute toxicity was evaluated in mice. Importantly, the effect of the CBF on lipid metabolism disorders was investigated in a high-fat diet (HFD) hyperlipidemia mouse model. An in silico study was carried out to predict underlying mechanisms. Results: The HPLC analysis revealed gallic acid, cinnamic acid, and naringenin as major phenolics of the carob pod aqueous extract. Concerning the basil hydro-ethanolic extract, rosmarinic, chicoric, caftaric, and caffeic acids were the main phenolics. Accordingly, the CBF prevented LRP oxidation in a concentration-dependent manner. This formula is not toxic in mice (LD₅₀ > 2000 mg/kg body weight). Moreover, animals administered the CBF at 200 mg/kg/day presented a significant decline in their body weight gain, adipose tissue weight, plasma total cholesterol, low-density lipoprotein cholesterol (LDL-C) level, and glycaemia after 10 weeks’ treatment. Accordingly, the CBF decreased the plasma atherogenic index and the LDL-C to HDL-C ratio and reduced the level of fats accumulated in the liver. The molecular docking study revealed that chicoric, rosmarinic, and caftaric acids, and naringenin bound particularly strongly to many proteins involved in the regulation of lipid and cholesterol metabolism. This includes the HMG-CoA reductase, PPARα/γ, PCSK9, Cyp7a1, and ATP-citrate lyase. Conclusions: The CBF could be a good source of natural supplements, functional foods, and pharmaceuticals effective in managing hyperlipidemia and oxidative stress and preventing their related cardiovascular disorders. Full article

Background: Oxaliplatin-induced neuropathy (OIN) is a severe painful condition that strongly affects the patient’s quality of life and cannot be counteracted by the available drugs or adjuvants. Thus, several efforts are devoted to discovering substances that can revert or reduce OIN, including natural compounds. The carob tree, Ceratonia siliqua L., possesses several beneficial properties. However, its antalgic properties have not been substantially investigated and only a few investigations have been conducted on the unripe carob (up-CS) pods. Thus, the aims of this study were to evaluate for the first time the unripe variety of Apulian carob, chemically characterized and profiled as antioxidant potential of polyphenolic compounds as well as to investigate the ability of up-CS to reduce the neurotoxicity in a mouse model of oxaliplatin-induced neuropathic pain. Methods: By UHPLC-HRMS/MS analyses, 50 phenolic compounds, belonging mainly to n-galloylated glucoses and flavonoids were detected. Results: In a mouse model of oxaliplatin-induced neurotoxicity (2.4 mg/kg, 10 injections over two weeks), acute per os treatment with up-CS provoked a dose-dependent pain-relieving effect that completely counteracted oxaliplatin hypersensitivity at the dose of 200 mg/kg. Repeated oral administration of up-CS (100 mg/kg), concomitantly with oxaliplatin injection, exerted a protective effect against the development of thermal and mechanical allodynia. In addition, up-CS exerted a neuroprotective role against oxaliplatin-induced astrocytes activation in the spinal cord measured as GFAP-fluorescence intensity. Conclusions: Overall, our study contributes to the knowledge on up-CS properties by highlighting its protective activity in the painful condition related to the administration of oxaliplatin. Full article

Dietary fiber (DF), and especially soluble dietary fiber (SDF), is a nutrient of particular interest today because of its anti-inflammatory role and its ability to reduce cardiovascular risk. Therefore, the enhancement of SDF in foods using different techniques has become a promising field of research. In order to prove the possibility of increasing this SDF content, the effects of different commercial enzymes (Pectinex^® Ultra SP-L, Viscozyme^® L and Celluclast^® 1.5 L) were tested on a variety of carob (CE), artichoke (ARE), apple (APE) and broccoli (BE) by-product extracts. Enzymatic treatment significantly affected SDF content in all by-products, showing the greatest increases for CE, ARE and APE using Celluclast^® 1.5 L, while Viscozyme^® L obtained the best results after application in BE. On the other hand, positive results were reported in the solubility, WHC and FAC of the by-products due to the enzymatic treatment, being increased in all extracts analyzed. Moreover, a general increase in antioxidant capacity (FRAP, ABTS and DPPH) was observed after enzymatic treatment. Finally, high yields were obtained after the application of the enzymatic processes, reaching values of 80–85% for each food by-product. These results evidenced a potential revalorization of carob, artichoke, apple and broccoli by-products after enzymatic treatment, improving its nutritional and physicochemical properties, revealing a possible application as a higher value-added ingredient. Full article

Carob pulp is a valuable source of cellulose-rich fraction (CRF) for many food applications. This study aimed to obtain and characterize a CRF derived from carob pulp waste after sugar removal and to evaluate its potential use in the 3D printing of cellulose-rich foods. Thus, the extraction of the CRF present in carob pulp (by obtaining the alcohol-insoluble residue) was carried out, accounting for nearly 45% dm (dry matter) of this byproduct. The CRF contained about 24% dm of cellulose. The functional properties (swelling capacity, water retention, and fat adsorption) related to this fraction were determined, showing a value of 5.9 mL/g of CRF and 4.0 and 6.5 g/g of CRF, respectively. Different gels were formulated with a total solids content of 15% wm (wet matter), using potato peel flour as a base and partially substituting with CRF (0% to 8% wm). The cellulose-based gels were characterized in terms of viscosity, water distribution (low-field Nuclear Magnetic Resonance), and printability, while the 3D printed samples were assessed for their textural properties. As the percentage of added CRF increased, the viscosity decreased while the water retention increased. Printability improved when small proportions of CRF (2% to 4%) were used, while it deteriorated for higher percentages (6% to 8%). The textural properties (hardness, adhesiveness, cohesiveness, and gumminess) showed significant changes caused by the addition of CRF, with gels containing 3% to 4% CRF exhibiting the most suitable printing values. In summary, this study demonstrates the significant potential of carob cellulose-based gel as an ingredient in the 3D printing of novel fiber-rich foods, contributing to reducing food waste and promoting sustainable practices within the framework of the circular economy. Full article

Carob leaves have gained attention for their bioactive properties and traditional medicinal uses, including as treatment for diabetes, digestive disorders, and microbial infections. The aim of this study was to explore the phytochemical composition of carob leaf acetone extracts using advanced spectroscopic techniques. The combined use of heteronuclear nuclear magnetic resonance (NMR) experiments with 1D selective nuclear Overhauser effect spectroscopy (NOESY) offers detailed structural insights and enables the direct identification and quantification of key bioactive constituents in carob leaf extract. In particular, the NMR and mass spectrometry techniques revealed the presence of myricitrin as a predominant flavonoid, as well as a variety of glycosylated derivatives of myricetin and quercetin, in acetone extract. Furthermore, siliquapyranone and related gallotannins are essential constituents of the extract. The potent inhibitory effects of the carob leaf extract on Staphylococcus aureus (MIC = 50 μg mL⁻¹) and a-glucosidase enzyme (IC₅₀ = 67.5 ± 2.4 μg mL⁻¹) were also evaluated. Finally, the antibacterial potency of carob leaf constituents were calculated in silico; digalloyl-parasorboside and gallic acid 4-O-glucoside exert a stronger bactericidal activity than the well-known myricitrin and related flavonoids. In summary, our findings provide valuable insights into the bioactive composition and health-promoting properties of carob leaves and highlight their potential for pharmaceutical and nutraceutical applications. Full article

The present study aimed to find solutions based on the use of plant-based ingredients that would improve the nutritional quality of meat products as well as ensure sensory and microbiological quality. Two fat replacers, lemon albedo (Citrus lemon) and carob seed gum (Ceratonia siliqua), were investigated by chemical analysis and panel testing to evaluate their effect on the nutritional and sensory quality of beef burgers. The antimicrobial activity of two plant extracts, from nettle (Urtica dioica) leaves and medlar (Eriobotrya japonica) seeds, was studied, evaluating the intensity of inhibitory action and the minimum inhibitory concentration against Pseudomonas spp. and Listeria innocua strains by plate test. In addition, the antioxidant activity of both extracts was evaluated. Based on the results, lemon albedo and medlar seed extracts were validated in a food model (beef burger) by a storage test and a challenge test. The storage test results highlight that medlar seed extract prevents the formation of thiobarbituric acid reactive substances (TBARSs) and ensures microbiological quality, inhibiting Enterobacteriaceae and Pseudomonas spp. Anti-Listeria efficacy was confirmed in situ by challenge test results. In conclusion, although fat replacers ensure nutritional and sensory quality, they do not satisfy microbiological quality. This study clearly demonstrates that the safety of low-fat burgers can only be achieved through the combination of appropriate fat replacers with well-selected natural antimicrobial extracts. Full article

Ceratonia siliqua L. is a medicinal plant that has long been used in traditional Moroccan medicine to treat many diseases. This study aimed to assess the impact of the stages of the immature phase of carob pulp (M1, M2, M3, M4, and M5) on phytochemical composition, antioxidant activity, and antidiabetic activity of Ceratonia siliqua L. The identification of the phenolic profile by HPLC-UV/MS-MS and the study of the antidiabetic effect by in silico, in vitro, and in vivo studies were carried out for extracts with high contents of phenolic compounds from immature wild carob pulp from the communes of Timoulit (TM), Bin Elouidane (AW), and Ouaouizerth (TG) in the province of Azilal in the Béni Mellal-Khénifra region. The results revealed a gradual increase in total sugar content over the pulp’s ripening period, reaching a value of 2134 ± 56.23 mg GE/100 g fresh weight (FW) for TG. The three locations showed peak values for total polyphenol content (TPC), total flavonoid content (TFC), and total condensed tannin (TCT) at the M2 stage. AW had the highest concentrations of TPC (3819 ± 226.4 mg GAE/100 g FM), TFC (1034 ± 57.08 mg QE/100 g FM), and TCT (1472 ± 28.46 mg CE/100 g FM). The DPPH assay (7892 ± 296.1 mg TE/100 g FM) and the FRAP assay (278.2 ± 7.85 mg TE/100 g FM) both demonstrated that the TG zone is a highly potent antioxidant zone. In contrast, the AW site exhibited a markedly elevated value of 725.4 ± 103.6 mg TE/100 g FM in the ABTS assay. HPLC-UV-MS/MS analysis showed that the methanolic extracts of immature carob pulp (MEICP) from the three areas contained several different chemical compounds. The most prevalent were 3-O-p-coumaroyl-5-O-caffeoylquinic acid, quercetin 3-methyl ether, gallic acid, and galloylquinic acid. Immature carob pulp extract (ICPE) from AW showed the strongest in vitro inhibition of pancreatic α-amylase (IC₅₀ = 0.405 µg/mL) and TG extracts were most potent against intestinal α-glucosidase (IC₅₀ = 0.063 µg/mL). In vivo, AW, TG, and TM extracts significantly reduced postprandial glycemia in rats, with AW having the greatest effect. These results highlight the antidiabetic potential of ICPE. The 3-O-p-Coumaroyl-5-O-caffeoylquinic acid showed better affinity for α-amylase compared to acarbose and interacted significantly with several amino acid residues of the enzyme. Similarly, this molecule and 3,4-Dicaffeoylquinic acid demonstrated a strong affinity for α-glucosidase, suggesting their potential as natural inhibitors of enzymes involved in carbohydrate metabolism. Most of the compounds are not substrates of P-glycoprotein and exhibited high intestinal absorption. Furthermore, the majority of these compounds did not act as inhibitors or substrates of CYP450 enzymes, reinforcing their suitability for development as oral medications. These results underscore the potential of immature carob pulp as a promising antidiabetic agent. Full article