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Structural characterization and in vitro lipid binding studies of non-specific lipid transfer protein 1 (nsLTP1) from fennel (Foeniculum vulgare) seeds.

ABSTRACT: Non-specific lipid transfer proteins (nsLTPs) are cationic proteins involved in intracellular lipid shuttling in growth and reproduction, as well as in defense against pathogenic microbes. Even though the primary and spatial structures of some nsLTPs from different plants indicate their similar features, they exhibit distinct lipid-binding specificities signifying their various biological roles that dictate further structural study. The present study determined the complete amino acid sequence, in silico 3D structure modeling, and the antiproliferative activity of nsLTP1 from fennel (Foeniculum vulgare) seeds. Fennel is a member of the family Umbelliferae (Apiaceae) native to southern Europe and the Mediterranean region. It is used as a spice medicine and fresh vegetable. Fennel nsLTP1 was purified using the combination of gel filtration and reverse-phase high-performance liquid chromatography (RP-HPLC). Its homogeneity was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry. The purified nsLTP1 was treated with 4-vinyl pyridine, and the modified protein was then digested with trypsin. The complete amino acid sequence of nsLTP1 established by intact protein sequence up to 28 residues, overlapping tryptic peptides, and cyanogen bromide (CNBr) peptides. Hence, it is confirmed that fennel nsLTP1 is a 9433 Da single polypeptide chain consisting of 91 amino acids with eight conserved cysteines. Moreover, the 3D structure is predicted to have four ?-helices interlinked by three loops and a long C-terminal tail. The lipid-binding property of fennel nsLTP1 is examined in vitro using fluorescent 2-p-toluidinonaphthalene-6-sulfonate (TNS) and validated using a molecular docking study with AutoDock Vina. Both of the binding studies confirmed the order of binding efficiency among the four studied fatty acids linoleic acid?>?linolenic acid?>?Stearic acid?>?Palmitic acid. A preliminary screening of fennel nsLTP1 suppressed the growth of MCF-7 human breast cancer cells in a dose-dependent manner with an IC₅₀ value of 6.98 µM after 48 h treatment.

SUBMITTER: Megeressa M

PROVIDER: S-EPMC7718255 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature

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Structural characterization and in vitro lipid binding studies of non-specific lipid transfer protein 1 (nsLTP1) from fennel (Foeniculum vulgare) seeds.

Megeressa Mekdes M Siraj Bushra B Zarina Shamshad S Ahmed Aftab A

Scientific reports 20201204 1

Non-specific lipid transfer proteins (nsLTPs) are cationic proteins involved in intracellular lipid shuttling in growth and reproduction, as well as in defense against pathogenic microbes. Even though the primary and spatial structures of some nsLTPs from different plants indicate their similar features, they exhibit distinct lipid-binding specificities signifying their various biological roles that dictate further structural study. The present study determined the complete amino acid sequence, ...[more]

PMID: 33277525

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Project description:Researchers recently focused on studying the nutritional and functional qualities of sprouts generated from seeds. The current study investigated the total phenolic content (TPC), total flavonoids (TF), total flavonols (TFL), antioxidant activity (AOA), specific phenolic acids, and volatile chemicals in fennel seeds (FS) and fennel seed sprouts (FSS). The oxidative DNA damage prevention activity of selected FS and FSS extracts against DNA was examined. Consequently, the antioxidative stress potential of FS and FSS extracts at 300 and 600 mg kg-1 on CCl4-induced hepatotoxicity and oxidative stress in rats weas investigated. The liver's functions and oxidative stress biomarkers in rat blood were examined. FSS exhibited rich phytochemical content such as TPC, TF, TFL, and AOA with altered phenolics and volatiles. HPLC identified nineteen compounds of phenolic acids and their derivatives in FS. Thirteen phenolics and six flavonoids were predominantly identified as Vanillic acid and Kaempferol, respectively. GC-MS analysis identified fifty and fifty-one components in FS and FSS, respectively. The predominant component was Benzene, [1-(2-propenyloxy)-3-butenyl] (trans-Anethole) (38.41%), followed by trans-Anethole (Benzene, 1-methoxy-4-(2-propenyl)) (23.65%), Fenchone (11.18%), and 1,7-Octadiene, 2-methyl-6-methylene- Cyclohexene (7.17%). Interestingly, α-Pinene, Fenchone, trans-Anethole (Benzene, 1-methoxy-4-(2-propenyl)), 4-Methoxybenzaldehyde (4-Anisaldehyde), Benzeneacetic acid, α-hydroxy-4-methoxy, and Nonacosane contents were increased. While Dillapiole, 7-Octadecenoic acid, and methyl ester were newly identified and quantified in FSS. The oxidative DNA damage prevention capability of FSS and FS extracts indicated remarkable DNA protection. Administrating FS and FSS extracts at 300 and 600 mg kg-1 ameliorated AST, ALT, and ALP, as well as GSH, CAT, MDA, and SOD, in a dose-dependent manner. The most efficient treatment of FS or FSS was using a dose of 600 mg Kg-1, which recorded an improvement rate of 20.77 and 24.17, 20.36 and 24.92, and 37.49 and 37.90% for ALT, AST, and ALP, respectively. While an improvement rate of 40.08 and 37.87%, 37.17 and 46.52%, 114.56 and 154.13%, and 66.05 and 69.69% for GSH, DMA, CAT, and SOD compared to the CCl4-group, respectively. The observed protection is associated with increased phenolics and volatiles in F. vulgare. Therefore, FS and FSS are recommended as functional foods with bioactive functionality, health-promoting properties, and desired prevention capabilities that may help prevent oxidative stress-related diseases.

Project description:BackgroundWith the increasing demands in livestock and poultry breeding and the growing number of food-borne diseases, it is necessary to practice food safety and develop strategies to produce healthy livestock. Fennel (Foeniculum vulgare Mill.) has been used as an additive in poultry production by some researchers, but there are few studies on the systemic beneficial effects of dietary fennel seed powder supplementation on broilers. Therefore, this study aimed to investigate the effect of dietary fennel seed powder supplementation on feed intake, the apparent metabolic rate of nutrients, intestinal morphology, and carcass traits in Cobb broilers.MethodsA single-factor experimental design was used. In total, 160 1-day-old Cobb broiler chicks were randomly assigned to four treatments, with four replicates each (n = 10/replicate). Broilers in the control (CN) group were fed a basal diet without fennel seed powder, and broilers in the treatment groups were fed a basal diet supplemented with 0.15% (LF), 0.30% (MF), or 0.45% (HF) fennel seed powder, respectively. Feeding trials lasted for 42 days under the conditions of ad libitum access to feed and water, and 24-h illumination. During the third and sixth weeks, digestive and metabolic assays were carried out. When the broilers were 42 days old, one chicken with a weight close to the average was selected from each repetition, euthanized by an intravenous injection of 5% sodium pentobarbital, and carcass traits were measured and intestinal samples were collected for morphological assessment.ResultsThere was no significant difference in growth performance of broilers (P > 0.05). The breast muscle percentage, fat width and fat width index, breast muscle area, and breast muscle area index of broilers in the LF group were higher than those in other groups (P < 0.05). Jejunum weight and length were higher in MF than in CN and LF broilers (P < 0.05). Additionally, duodenal villi height, ileal villi height, and ileal wall thickness were higher in MF than in CN broilers (P < 0.05). There were no significant differences in nutrient utilization among all groups (P > 0.05), except that the ash apparent metabolic rate in MF broilers at 21 days of age was higher than that in LF broilers (P < 0.05). In conclusion, dietary supplementation with a moderate concentration of fennel affects carcass performance, and intestinal morphology, and promotes the growth and development of broilers.

Dataset Information

Structural characterization and in vitro lipid binding studies of non-specific lipid transfer protein 1 (nsLTP1) from fennel (Foeniculum vulgare) seeds.

Publications

Structural characterization and in vitro lipid binding studies of non-specific lipid transfer protein 1 (nsLTP1) from fennel (Foeniculum vulgare) seeds.

Similar Datasets

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