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AbstractPeptide-based medications hold immense potential in addressing a wide range of human disorders and discomforts. However, their widespread utilization encounters two major challenges: preservation and production efficiency. Cyclotides, a class of ribosomally synthesized and post-translationally modified peptides (RiPPs), exhibit unique characteristics, such as a cyclic backbone and cystine knot, enhancing their stability and contributing to a wide range of pharmacological properties exhibited by these compounds. Cyclotides are efficient in the biomedical (e.g., antitumor, antidiabetic, antimicrobial, antiviral) and agrochemical fields by exhibiting activity against pests and plant diseases. Furthermore, their structural attributes make them suitable as molecular scaffolds for grafting and drug delivery. Notably, the mutated variant of kalata B1 cyclotide ([T20K] kalata B1) has recently entered phase 1 of human clinical trials for multiple sclerosis, building upon the success observed in animal trials. To enable large-scale production of cyclotides, it is crucial to further explore their remarkable structural and bioactive properties. This necessitates extensive research focused on enhancing the efficiency of the processes required for their production. This study provides a comprehensive review of the biological synthesis methods of cyclotides, with particular emphasis on various expression systems, namely bacteria, plants, yeast, and cell-free systems. By investigating these expression systems, it becomes possible to design production systems that are adaptable, economically viable, and efficient for generating active and pure cyclotides at an industrial scale. The advantages of biological synthesis over chemical synthesis are thoroughly explored, highlighting the potential of these expression systems in meeting the demands of large-scale cyclotide production. [ABSTRACT FROM AUTHOR]

The cholecystokinin-2 receptor (CCK2R) is a G protein-coupled receptor (GPCR) that is expressed in peripheral tissues and the central nervous system and constitutes a promising target for drug development in several diseases, such as gastrointestinal cancer. The search for ligands of this receptor over the past years mainly resulted in the discovery of a set of distinct synthetic small molecule chemicals. Here, we carried out a pharmacological screening of cyclotide-containing plant extracts using HEK293 cells transiently-expressing mouse CCK2R, and inositol phosphate (IP1) production as a readout. Our data demonstrated that cyclotide-enriched plant extracts from Oldenlandia affinis, Viola tricolor and Carapichea ipecacuanha activate the CCK2R as measured by the production of IP1. These findings prompted the isolation of a representative cyclotide, namely caripe 11 from C. ipecacuanha for detailed pharmacological analysis. Caripe 11 is a partial agonist of the CCK2R (Emax = 71%) with a moderate potency of 8.5 µM, in comparison to the endogenous full agonist cholecystokinin-8 (CCK-8; EC50 = 11.5 nM). The partial agonism of caripe 11 is further characterized by an increase on basal activity (at low concentrations) and a dextral-shift of the potency of CCK-8 (at higher concentrations) following its co-incubation with the cyclotide. Therefore, cyclotides such as caripe 11 may be explored in the future for the design and development of cyclotide-based ligands or imaging probes targeting the CCK2R and related peptide GPCRs. [ABSTRACT FROM AUTHOR]

Natural compounds are proper tools for inhibiting cancer cell proliferation. Hence, the search for these ligands of overexpressed receptors in breast cancer has been a competitive challenge recently and opens new avenues for drug discovery. In this research, we have investigated molecular interactions between natural products and overexpressed receptors in breast cancer using molecular docking and dynamic simulation approaches followed by extraction of the best ligand from Citrus limetta and developing for nanoscale encapsulation composed of soy lecithin using a sonicator machine. The encapsulation process was confirmed by DLS and TEM analyses. Anticancer activity was also examined using MTT method. Among the investigated natural compounds, hesperidin was found to bind to specific targets with stronger binding energy. The molecular dynamics results indicated that the hesperidin-MCL-1 complex is very stable at 310.15 K for 200 ns. The RP-HPLC analysis revealed that the purity of extracted hesperidin was 98.8% with a yield of 1.72%. The results of DLS and TEM showed a strong interaction between hesperidin and lecithin with an entrapped efficiency of 92.02 ± 1.08%. Finally, the cytotoxicity effect of hesperidin was increased against the MDA-MB-231 cell line with an IC50 value of 62.93 μg/mL after encapsulation, whereas no significant effect against the MCF10A cell line. We showed for the first time that hesperidin is a flexible and strong ligand for the MCL-1 receptor. Also, it has the in vitro ability to kill the MDA-MB-231 cell lines without having a significant effect on the MCF10A cell lines. Therefore, hesperidin could be used as a food ingredient to generate functional foods. [ABSTRACT FROM AUTHOR]

Functional evaluation of encrypted bioactive peptides in protein structure helps to better understand those for using in pharmacy and food sciences. For this purpose, the total protein was extracted from Matricaria chamomilla, Ziziphora clinopodioides, and Cressa cretica, and partially purified with ammonium sulfate. Protein hydrolysates were obtained from pancreatin hydrolysis for 240 min and the enzyme hydrolysis was confirmed using the determination of hydrolysis degree and Fourier transform infrared (FT‐IR) followed by the physicochemical and sensory properties were investigated. The results showed that all hydrolysates had both cytotoxic and antioxidant activities. Specifically, C. cretica hydrolysates represented cytotoxic activity against the MCF‐7 cell line with the IC50 of 135.21 µg/mL, while showed no significant growth inhibition effect on the HEK293 cell line. Besides, M. chamomilla hydrolysates showed the lowest bitterness value (1.125 ± 0.52). From the perspective of color investigation, M. chamomilla hydrolysates indicated the highest L* and the lowest a* factors. The highest turbidity and surface tension, and 10‐fold more cancer cell killing effect under gastrointestinal digestion conditions were observed for M. chamomilla hydrolysates. Therefore, bioactive peptides might be formulated in designing of novel anticancer drugs or could be used in promising protocols for the production of food products with beneficial health effects. [ABSTRACT FROM AUTHOR]

[Display omitted] • The peptides inhibit the growth of MCF-7 cell line strongly. • The results of antioxidant activities showed that the peptides have a high effect. • Three major peaks detected using LC–MS, which were a dipeptide or tripeptide. • The SEM analysis showed that these peptides create pores in the MCF-7 cell membrane. Despite the limitations of current methods in cancer treatment, the use of bioactive peptides can be as an alternative to treat today. Therefore, isolation and relative purification of bioactive peptides was carried out form Achillea eriophora using a Sep-Pak C18 SPE cartridge and Amicon® Ultra Centrifugal Filters. The presence of desired peptides was checked using RP-HPLC and confirmed using LC-MS. The results of anticancer assay showed that the peptide mixture inhibits the growth of MCF-7 cancerous cell line with the values of IC 50 , GI 50 , and LC 50 equal to 18.73 ± 0.22, 7.52 ± 0.15, and 56.73 ± 0.18 µg/mL, respectively. It also showed DPPH radical scavenging activity and cupric-ion reducing power with the IC 50 value of 5.095 ± 0.23 and 63.3 ± 0.44 µg/mL, respectively. Although flavonoids were present in the sample along with the peptides, their amount was trivial (18.097 ± 1.36 μg/mL). Nevertheless, the results of the LC-MS showed mass-to-charge ratios of 301.17, 261.22, and 243.25, which was a dipeptide or tripeptide in compression to enzyme-digested BSA as a standard. In addition, SEM analysis of the purified peptide mixture showed that it kills the MCF-7 cancerous cell line by creating pores in the membrane. Therefore, it might be valuable to these peptides sequenced and be studied for physicochemical properties. Animal and clinical studies could help its application in drug development. [ABSTRACT FROM AUTHOR]

Camelina sativa oil has garnered significant interest due to its potential as a source of natural bioactive compounds. This study focuses on optimizing the extraction of C. sativa oil and evaluating its cytotoxic effects against MCF-7 breast cancer cells using response surface methodology (RSM). The optimal extraction conditions were determined to be a ratio of 9:1 mL/g, with an extraction time of 13.78 min at 60.06℃ for oil extraction (11.69 %, p-value ≤ 0.05), and also a ratio of 6.5:1 mL/g with a time of 10.64 min at 70.18℃ for cytotoxic effect (63.80 %, p-value ≤ 0.05). Gas chromatography analysis identified key components of the oil, including oleic acid (23.44 %), 8,11-octadecadienoic acid (13.48 %), and palmitic acid (6.31 %). The physical properties of the oil, such as refractive index (1.469), density (0.9132 g/cm³), and viscosity (0.9103 g/cm³) were also characterized. The extracted oil displayed antioxidant activity with a DPPH IC 50 value of 78.40 µg/mL. Molecular docking studies revealed stable interactions between the oil components and critical residues in the active sites of estrogen and progesterone receptors linked to breast cancer. ADMET analysis indicated compliance with Lipinski's Rule of Five, although varying solubility and absorption characteristics were noted. Overall, C. sativa oil exhibited promise as a dual-functioning agent with both anticancer and antioxidant properties, highlighting the necessity for further research to confirm its therapeutic potential. [Display omitted] • Oil extraction and its cytotoxicity effect was optimized using RSM. • Oil key compounds included oleic acid, 8,11-octadecadienoic acid, and palmitic acid. • Refractive index, density, and viscosity of oil were of interest in the study. • The extracted oil exhibited DPPH radical scavenging effect (IC 50 = 78.40 µg/mL). • Oil compounds interacted with breast cancer-linked receptors with high affinity. [ABSTRACT FROM AUTHOR]

Conversion of manures to vermicompost and biochar may alleviate some negative effects of manure application to soil but the efficiency of the produced vermicompost and biochar as compared to their feedstocks is not well-known. In the current investigation, we compared the effects of sheep manure and its derived vermicompost and biochar (pyrolyzed at 400°C for 4 h) on the properties of a calcareous soil that planted with five cultivars of barley (Behrokh, Khatam, Reyhaneh03, Fajr 30 and Nimrooz) for 60 days. Different soil properties and availability of nutrients and barley yield were determined after plant harvest. The biochar significantly increased barley yield rather than control (4.20 vs. 3.57 g pot−1), but sheep manure and vermicompost had no effect on it (3.51 and 3.37 g pot−1, respectively). Fajr 30 and Nimrooz (3.52 and 3.42 g pot−1, respectively) had significantly lower yield than other cultivars. Biochar increased soil pH up to 8.2. Soil salinity was increased by application of all organic materials (increase to 16–36%). Cation exchange capacity (CEC) and organic matter content of soil were also increased by all organic materials application (0.4–0.9 cmol kg−1 and 0.33–0.50%, respectively). All organic materials increased total nitrogen (N), but this increase was the highest with sheep manure application (53%). The availability of phosphorus (P) and potassium (K) was increased significantly by application of all organic materials, and this increase was the highest with biochar application (19 and 309 mg kg−1, respectively). Biochar application had no effect on the availability of micronutrients, but application of sheep manure and vermicompost increased the availability of iron (Fe) (0.62 and 0.48 mg kg−1, respectively) and zinc (Zn) (0.18 and 0.37 mg kg−1, respectively). Generally, organic materials may change the status of soil nutrients via change in soil pH, organic matter content, release of nutrients, increase in soil CEC and formation of soluble complex with nutrients. [ABSTRACT FROM AUTHOR]

The effects of ultrasound‐assisted extraction (UAE) and microwave‐assisted extraction (MAE) methods on molecular and physicochemical characteristics of the resultant gelatin were examined. Before extraction procedure, we investigated the optimum pH for swelling of Common carp by‐products, which is an important pretreatment for gelatin production. The highest swelling yield was achieved at pH 13 among pH 1–14 with unit intervals. Results indicated that the UAE gelatin has a higher gel strength, viscosity, melting point, and gelling point. The power and time of sonication showed a reverse relation with these characteristics. In addition, as the time of microwave heating was raised, the gel strength, viscosity, melting point, and gelling point were decreased. The FT‐IR spectra showed similar peaks but the Amide B in UAE gelatin slightly vanished. The electrophoretic pattern also revealed the higher gel strength and viscosity of UAE gelatin due to the higher intensity of α and β chains compared to MAE gelatin. It can be concluded from all of the results of this study that the produced gelatin using these procedures can be a good source of gelatin in food and drug industries. [ABSTRACT FROM AUTHOR]

Medicinal plants contain valuable compounds that have attracted worldwide interest for their use in the production of natural drugs. The presence of compounds such as rosmarinic acid, carnosic acid, and carnosol in Rosmarinus officinalis has made it a plant with unique therapeutic effects. The identification and regulation of the biosynthetic pathways and genes will enable the large-scale production of these compounds. Hence, we studied the correlation between the genes involved in biosynthesis of the secondary metabolites in R. officinalis using proteomics and metabolomics data by WGCNA. We identified three modules as having the highest potential for the metabolite engineering. Moreover, the hub genes highly connected to particular modules, TFs, PKs, and transporters were identified. The TFs of MYB, C3H, HB, and C2H2 were the most likely candidates associated with the target metabolic pathways. The results indicated that the hub genes including Copalyl diphosphate synthase (CDS), Phenylalanine ammonia lyase (PAL), Cineole synthase (CIN), Rosmarinic acid synthase (RAS), Tyrosine aminotransferase (TAT), Cinnamate 4-hydroxylase (C4H), and MYB58 are responsible for biosynthesis of important secondary metabolites. Thus, we confirmed these results using qRT-PCR after treating R. officinalis seedlings with methyl jasmonate. These candidate genes may be employed for genetic and metabolic engineering research to increase R. officinalis metabolite production. [ABSTRACT FROM AUTHOR]

This study aims to research the impact of coatings containing whey protein (WP), fish gelatin hydrolysates (FGH), and both compounds together (WP + FGH) on the shelf-life of chicken breast fillets over the course of 16 days of cold storage (4 °C, 4-day intervals), as assessed by their physicochemical, microbiological, and sensory properties. Overall, cooking loss, pH value, total volatile base nitrogen, free fatty acids, peroxide value, and thiobarbituric acid reactive substances increased with storage time in all samples. WP + FGH coated samples had significantly lower variation in all these parameters over the time of storage compared to other coated samples (WP and FGH), while these parameters increased greatly in control (uncoated) samples. WP + FGH coating also resulted in reduced bacterial counts of total mesophilic, aerobic psychrotrophic, and lactic acid bacteria compared to other coated and uncoated samples. The sensory evaluation revealed no differences in the panelists' overall acceptance at day 0 of storage between samples. The samples were considered "non-acceptable" by day 8 of storage; however, WP + FGH coated samples maintained an overall higher acceptability score for the sensory attributes evaluated by the panelists. Overall, this study shows the potential of WP + FGH coatings for prolonging the shelf-life of chicken breast fillets. [ABSTRACT FROM AUTHOR]

Cecropin A, as an antimicrobial peptide (AMP), is possible to use in medical and agricultural fields as a new and safe biocontrol agent. Therefore, it is highly necessary to find a cost‐effective and scalable approach to generate a large scale of it. In this research, the Agrobacterium rhizogenes strain ATCC 15834 was used to transfer the Cecropin A gene to the Nicotiana tabacum. After confirmation of transgenic hairy roots, the antibacterial activity of purified Cecropin A peptide was measured using the agar gel diffusion method. Successful transforming of Cecropin A was confirmed at the RNA and protein levels in hairy root cells using RT‐PCR and enzyme‐linked immunosorbent assay (ELISA), respectively. The highest Cecropin A amount was detected in line 4 of the transgenic lines using ELISA in comparison with the nontransgenic line. Subsequently, the antimicrobial activity of Cecropin A extracted from line 4 showed the highest inhibition activity against Aspergillus niger. Besides, this activity was stable against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Candida albicans pathogens after 7 days. The recombinant production of Cecropin A AMP had a yield of 63.81 μg/g of fresh weight. According to a significant yield, this system can be used to produce the Cecropin A peptide for pharmacological and food science applications. [ABSTRACT FROM AUTHOR]