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1.
World J Microbiol Biotechnol ; 40(5): 152, 2024 Mar 30.
Article in English | MEDLINE | ID: mdl-38553646

ABSTRACT

Saponins are a large group of compounds, produced mostly by plants as a side product of their metabolic activity. These compounds have attracted much attention over the years mostly because of their surface activity and antibacterial, anti-inflammatory and antifungal properties. On the other hand, most of the hitherto research has concerned the action of saponins against microbial cells as a whole. Therefore, knowing the possible interaction of saponins with biomembrane, we decided to check in-vitro the influence of saponin-rich extract of Saponaria officinalis on spheroplasts of two Candida sp. The obtained results show that 10 mg L- 1 of extract increased the permeability of spheroplasts up to 21.76% relative to that of the control sample. Moreover, the evaluation of surface potential has revealed a decrease by almost 10 mV relative to that of the untreated samples. Such results suggest its direct correlation to integration of saponins into the biomembrane structure. The obtained results have proved the antifungal potential of saponins and their ability of permeabilization of cells. This proves the high potential of saponins use as additives to antifungal pharmaceutics, which is expected to lead to improvement of their action or reduction of required dosage.


Subject(s)
Saponaria , Saponins , Antifungal Agents/pharmacology , Antifungal Agents/chemistry , Saponaria/chemistry , Saponins/pharmacology , Saponins/chemistry , Plant Extracts/pharmacology , Plant Extracts/chemistry , Candida , Permeability
2.
FASEB J ; 36(7): e22387, 2022 07.
Article in English | MEDLINE | ID: mdl-35696068

ABSTRACT

Targeting Toll-like receptor 4/myeloid differentiation factor 2 (TLR4/MD2) signaling is regarded as a potential strategy for treating inflammatory diseases. Saponaria officinalis L. is rich in saponin, which include quillaic acid, gypsogenin, saponarin, and hederagenin. We evaluated the pharmacological activity of a Saponaria officinalis extract in THP-1 derived macrophages and RAW264.7 macrophages. TLR4/MyD88 complex formation and downstream signals were investigated by co-immunoprecipitation (Co-IP). In silico docking simulation was conducted to predict binding scores and perform 3D modeling of saponarin-TLR4/MD2 complex. A hexane fraction of Saponaria officinalis (SH) and fr.1 (a sub-fraction 1 of SH) inhibited mitogen-activated protein kinase (MAPK) signaling, nuclear factor kappa b (NF-κB) activity, cytokine production, and the expressions of marker genes specific for M1 polarization. The inhibitory effects of fr.1 and saponarin on TLR4/MyD88 complex formation were observed by western blotting TLR4 co-immunoprecipitated proteins. Saponarin and fr.1 markedly attenuated LPS-induced inflammatory cytokines, thus reducing mortality and morphological abnormality in zebrafish larvae. Finally, docking simulation revealed that saponarin can directly interact with TLR4/MD2 complex to inhibit downstream signalings. Our findings suggest that saponarin reduces downstream inflammatory response by disrupting TLR4/MD2 complex and blocking MyD88-dependent inflammatory signaling.


Subject(s)
Saponaria , Toll-Like Receptor 4 , Adaptor Proteins, Signal Transducing/metabolism , Animals , Lipopolysaccharides/pharmacology , Myeloid Differentiation Factor 88/metabolism , NF-kappa B/metabolism , Saponaria/metabolism , Signal Transduction/physiology , Toll-Like Receptor 4/metabolism , Zebrafish/metabolism , Zebrafish Proteins/metabolism
3.
Int J Mol Sci ; 24(14)2023 Jul 20.
Article in English | MEDLINE | ID: mdl-37511452

ABSTRACT

Saponaria sicula Raf. grows in Sicily, Sardinia, and Algeria on limestone cliffs and volcanic sands 1300-2500 m above sea level. The aim of the present study was to investigate how the pedo-climatic conditions influence the micromorphological, phytochemical, and biological properties of Sicilian S. sicula leaves collected in the Madonie Mountains (SsM) and on Etna Mt (SsE). Micromorphological investigations revealed that leaves from SsM had a higher amount of calcium oxalate druses in the mesophyll and a more intense blue-green staining with Toluidine blue O, indicating a higher content of polyphenols. These data were confirmed by phytochemical analyses carried out on hydroalcoholic extracts, which showed a higher content of total phenols (8.56 ± 0.57 g GAE/100 g DE) and flavonoids (6.09 ± 0.17 g RE/100 g DE) in SsM. Sixty-four compounds were identified by LC-DAD-ESI-MS analysis with propelargonidin dimer as the most abundant compound (10.49% and 10.19% in SsM and SsE, respectively). The higher polyphenol content of SsM leaves matches also with their biological activity, identifying SsM extract as the strongest plant complex (IC50 2.75-477.30 µg/mL). In conclusion, the present study experimentally demonstrates that not only climatic differences but also soil characteristics affect the micromorphological, phytochemical, and biological features of this plant species.


Subject(s)
Saponaria , Antioxidants/analysis , Plant Extracts/chemistry , Polyphenols/analysis , Flavonoids/analysis , Phytochemicals/chemistry , Plant Leaves/chemistry , Sicily
4.
Int J Mol Sci ; 23(4)2022 Feb 12.
Article in English | MEDLINE | ID: mdl-35216169

ABSTRACT

Saponaria officinalis L., commonly known as "Soapwort", is a rich source of triterpene glycosides; however, the chemical constituents of S. officinalis seeds have not been fully identified. In this study, we conducted a systematic phytochemical investigation of the seeds of S. officinalis and obtained 17 oleanane-type triterpene glycosides (1-17), including seven new glycosides (1-7). The structures of 1-7 were determined based on a detailed analysis of NMR spectroscopic data and chromatographic and spectroscopic analyses following specific chemical transformation. The cytotoxicities of the isolated compounds were evaluated against HL-60 human promyelocytic leukemia cells, A549 human adenocarcinoma lung cancer cells, and SBC-3 human small-cell lung cancer cells. The cytotoxicities of 1, 4, and 10 toward HL-60 cells and SBC-3 cells were nearly as potent as that of cisplatin. Compound 1, a bisdesmosidic triterpene glycoside obtained in good yield, arrested the cell cycle of SBC-3 cells at the G2/M phase, and induced apoptosis through an intrinsic pathway, accompanied by ROS generation. As a result of the mitochondrial dysfunction induced by 1, mitochondria selective autophagy, termed mitophagy, occurred in SBC-3 cells.


Subject(s)
Antineoplastic Agents/toxicity , Apoptosis , Mitochondria/metabolism , Oleanolic Acid/toxicity , Saponaria/chemistry , A549 Cells , Cell Cycle/drug effects , Humans , Oleanolic Acid/metabolism , Saponaria/metabolism , Seeds/chemistry , Seeds/metabolism
5.
Molecules ; 27(18)2022 Sep 08.
Article in English | MEDLINE | ID: mdl-36144548

ABSTRACT

The purpose of this study was to identify the chemical components in root extracts of Saponaria cypria, an endemic species of Cyprus. Subsequently, the synergistic bioactivity of its root extracts through different extraction procedures was also investigated for the first time. A total of nine saponins, along with six phenolic compounds, were identified and quantified using the UHPLC/Q-TOF-MS method. Additionally, S. cypria root extracts demonstrated antibacterial potential against Escherichia coli, Staphylococcus aureus, Enterococcus faecalis and Salmonella enteritidis. S. aureus presented the highest susceptibility among all bacteria tested. These findings provide the first phytochemical data regarding the saponin, phenolic content and antimicrobial activity of S. cypria extracts, indicating that the Cyprus saponaria species is a rich natural source for bioactive compounds with a potentially wider bioactivity spectrum.


Subject(s)
Saponaria , Saponins , Anti-Bacterial Agents/pharmacology , Microbial Sensitivity Tests , Phenols/pharmacology , Phytochemicals , Plant Extracts/chemistry , Plant Extracts/pharmacology , Saponaria/chemistry , Saponins/pharmacology , Staphylococcus aureus
6.
Bioorg Chem ; 113: 105032, 2021 08.
Article in English | MEDLINE | ID: mdl-34089947

ABSTRACT

This study attempts to evaluate the antioxidant, enzyme inhibitory, and anticancer properties as well as fatty acid compositions of endemic Saponaria prostrata WILLD. subsp. anatolica HEDGE. The gas chromatography-mass spectrometry (GC-MS) was used to determine the fatty acid content of methanol: dichloromethane extract from S. prostrata subsp. anatolica (SPA). Enzymatic activity was measured against acetylcholinesterase, butyrylcholinesterase and α-glucosidase. DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging activity and Ferric reducing antioxidant power assay (FRAP) were conducted to antioxidant properties. The anticancer effect of SPA on human MCF-7 breast cancer and human HCT116 colorectal cancer cell line was evaluated by WST-1 cell viability assay, colony formation assay and wound healing assay. In addition, human VEGF Elisa method was used to determine the anti-angiogenic effect, and the quantitative real-time PCR (qRT-PCR) method on p53, Bax and Bcl-2 mRNA levels were used to evaluate apoptosis. While high amounts of palmitic acid (40.8%), linoleic acid (17.75%) and α-linolenic acid (16.84%) were detected in the SPA, the total amount of unsaturated fatty acid (51.34%) was higher than the total amount of saturated fatty acid (48.66%). SPA displayed the most promising acetylcholinesterase (AChE), butyrylcholinesterase (BuChE) and α-glycosidase (AG) inhibitory activities (AChE: IC50: 18.03 µg/mL, BuChE: IC50: 44.24 µg/mL and AG: IC50: 210.85 µg/mL). The half maximum inhibitory concentration (IC50) of SPA in MCF-7 and HCT116 cells was determined as 259.79 µg/mL and 97.24 µg/mL, respectively. In addition, it was determined that SPA suppresses colony formation and wound closure, and suppresses angiogenesis as well as triggering apoptosis at a significant level. It is true that endemic S. prostrata subsp. anatolica is a potential source of functional food ingredients, but more analytical and in vivo experiments are needed to explore further secondary metabolite diversity and pharmacological properties.


Subject(s)
Antineoplastic Agents, Phytogenic/chemistry , Antioxidants/chemistry , Fatty Acids/analysis , Plant Extracts/chemistry , Saponaria/chemistry , Acetylcholinesterase/chemistry , Acetylcholinesterase/metabolism , Angiogenesis Inhibitors/chemistry , Angiogenesis Inhibitors/metabolism , Angiogenesis Inhibitors/pharmacology , Antineoplastic Agents, Phytogenic/metabolism , Antineoplastic Agents, Phytogenic/pharmacology , Apoptosis/drug effects , Butyrylcholinesterase/chemistry , Butyrylcholinesterase/metabolism , Cell Line, Tumor , Cell Movement/drug effects , Cell Survival/drug effects , Cholinesterase Inhibitors/chemistry , Cholinesterase Inhibitors/metabolism , Glycoside Hydrolase Inhibitors/chemistry , Glycoside Hydrolase Inhibitors/metabolism , Humans , Saponaria/metabolism , alpha-Glucosidases/chemistry , alpha-Glucosidases/metabolism
7.
Molecules ; 26(1)2021 Jan 05.
Article in English | MEDLINE | ID: mdl-33466379

ABSTRACT

We evaluated a hydroalcoholic extract of Sapindus saponaria L. pericarps (ETHOSS), as a candidate to a topical antifungal medicine for onychomycosis. ETHOSS was produced by extracting the crushed fruits in ethanol. The saponin contents were identified and characterized by electrospray ionization mass spectrometry. We measured the in vitro antifungal activity against three dermatophyte fungi, isolated from onychomycosis: Trichophyton rubrum, T. mentagrophytes, and T. interdigitale, using broth microdilution tests. The minimum fungicide concentration of ETHOSS ranged from 195.31 to 781.25 µg/mL. The cytotoxicity of the crude extract was tested on the HeLa cell line, and its ability to permeate into healthy human nails by photoacoustic spectroscopy and Fourier transformation infrared spectrometer (FTIR) spectroscopy by attenuated total reflection. Besides its strong antifungal activity, ETHOSS showed low cytotoxicity in human cells. It was able to permeate and reach the full thickness of the nail in one hour, without the aid of facilitating vehicles, and remained there for at least 24 h. These results suggest that ETHOSS has great potential for treating onychomycosis.


Subject(s)
Alcohols/chemistry , Antifungal Agents/pharmacology , Nails/drug effects , Plant Extracts/pharmacology , Saponaria/chemistry , Saponins/pharmacology , Adult , Female , Humans , Nails/metabolism
8.
Molecules ; 26(7)2021 Apr 04.
Article in English | MEDLINE | ID: mdl-33916520

ABSTRACT

In the present work, the properties of ZnO nanoparticles obtained using an eco-friendly synthesis (biomediated methods in microwave irradiation) were studied. Saponaria officinalis extracts were used as both reducing and capping agents in the green nanochemistry synthesis of ZnO. Inorganic zinc oxide nanopowders were successfully prepared by a modified hydrothermal method and plant extract-mediated method. The influence of microwave irradiation was studied in both cases. The size, composition, crystallinity and morphology of inorganic nanoparticles (NPs) were investigated using dynamic light scattering (DLS), powder X-ray diffraction (XRD), SEM-EDX microscopy. Tunings of the nanochemistry reaction conditions (Zn precursor, structuring agent), ZnO NPs with various shapes were obtained, from quasi-spherical to flower-like. The optical properties and photocatalytic activity (degradation of methylene blue as model compound) were also investigated. ZnO nanopowders' antibacterial activity was tested against Gram-positive and Gram-negative bacterial strains to evidence the influence of the vegetal extract-mediated synthesis on the biological activity.


Subject(s)
Anti-Bacterial Agents/pharmacology , Metal Nanoparticles/chemistry , Saponaria/chemistry , Zinc Oxide/pharmacology , Anti-Bacterial Agents/chemical synthesis , Candida albicans/drug effects , Candida albicans/growth & development , Catalysis , Escherichia coli/drug effects , Escherichia coli/growth & development , Green Chemistry Technology , Humans , Light , Metal Nanoparticles/ultrastructure , Microbial Sensitivity Tests , Microwaves , Photochemical Processes , Pseudomonas aeruginosa/drug effects , Pseudomonas aeruginosa/growth & development , Staphylococcus aureus/drug effects , Staphylococcus aureus/growth & development , Zinc Oxide/chemistry
9.
Molecules ; 26(18)2021 Sep 16.
Article in English | MEDLINE | ID: mdl-34577098

ABSTRACT

Our skin is continuously exposed to different amphiphilic substances capable of interaction with its lipids and proteins. We describe the effect of a saponin-rich soapwort extract and of four commonly employed synthetic surfactants: sodium lauryl sulfate (SLS), sodium laureth sulfate (SLES), ammonium lauryl sulfate (ALS), cocamidopropyl betaine (CAPB) on different human skin models. Two human skin cell lines were employed: normal keratinocytes (HaCaT) and human melanoma cells (A375). The liposomes consisting of a dipalmitoylphosphatidylcholine/cholesterol mixture in a molar ratio of 7:3, mimicking the cell membrane of keratinocytes and melanoma cells were employed as the second model. Using dynamic light scattering (DLS), the particle size distribution of liposomes was analyzed before and after contact with the tested (bio)surfactants. The results, supplemented by the protein solubilization tests (albumin denaturation test, zein test) and oil emulsification capacity (using olive oil and engine oil), showed that the soapwort extract affects the skin models to a clearly different extent than any of the tested synthetic surfactants. Its protein and lipid solubilizing potential are much smaller than for the three anionic surfactants (SLS, ALS, SLES). In terms of protein solubilization potential, the soapwort extract is comparable to CAPB, which, however, is much harsher to lipids.


Subject(s)
Biomimetics/methods , Plant Extracts/chemistry , Saponaria/chemistry , Skin/drug effects , Surface-Active Agents/chemistry , 1,2-Dipalmitoylphosphatidylcholine/chemistry , Betaine/analogs & derivatives , Betaine/chemistry , Cell Line , Cell Survival/drug effects , Cholesterol/chemistry , Emulsifying Agents/chemistry , Humans , Hydrogen-Ion Concentration , Keratinocytes/drug effects , Liposomes/chemistry , Models, Biological , Particle Size , Saponins/chemistry , Sodium Dodecyl Sulfate/analogs & derivatives , Sodium Dodecyl Sulfate/chemistry , Triterpenes/chemistry , Zein/chemistry
10.
Molecules ; 26(24)2021 Dec 20.
Article in English | MEDLINE | ID: mdl-34946786

ABSTRACT

Saponins are an important group of secondary metabolites naturally occurring in plants with important properties like: antibacterial, antiviral and antifungal. Moreover, they are widely used in the cosmetic industry and household chemistry. The sapogenins are saponin hydrolyses products, frequently used to facilitate saponin detection. In the present study, an improved methodology for isolation and separation of five sapogenins extracted from nettle (Urtica dioica L.), white dead-nettle (Lamium album L.), common soapwort (Saponaria officinalis L.) and washnut (Sapindus mukorossi Gaertn.) was developed using ultra-high-performance liquid chromatography with an evaporative light-scattering detector (UHPLC-ELSD). Based on quantitative analysis, the highest content of hederagenin (999.1 ± 6.3 µg/g) and oleanolic acid (386.5 ± 27.7 µg/g) was found in washnut extracts. Good recoveries (71% ± 6 up to 99% ± 8) were achieved for four investigated targets, while just 22.2% ± 0.5 was obtained for the fifth one. Moreover, hederagenin and oleanolic acid of whose highest amount was detected in washnut (999.1 ± 6.3 µg/g and 386.5 ± 27.7 µg/g, respectively) were subject to another approach. Consequently, liquid chromatography coupled mass spectrometry (LC/MS) with multiple reaction monitoring mode (MRM) was used as an additional technique for fast and simultaneous identification of the mentioned targets.


Subject(s)
Sapindus/chemistry , Sapogenins/analysis , Sapogenins/isolation & purification , Saponaria/chemistry , Urtica dioica/chemistry
11.
J Sci Food Agric ; 101(2): 786-793, 2021 Jan 30.
Article in English | MEDLINE | ID: mdl-32869316

ABSTRACT

BACKGROUND: This study evaluated the usability of saponin-rich extracts (soapwort and horse chestnut) as a foaming agent for foam mat drying of pomegranate juice. RESULTS: According to the foaming and stabilization studies, the optimum conditions were determined as 0.4% of soapwort extract, 0.03% of carboxymethyl cellulose as a stabilizer, and 3 min of whipping time. The foams produced using these conditions were dried at different spreading thicknesses and drying temperatures. The results showed that the thicker spreading thicknesses provided a higher antioxidant activity. On the other hand, drying temperature had a significant effect on all measured parameters except moisture content and water activity. The higher drying temperature caused a greater colour change and a lower content of total phenolics, total monomeric anthocyanins, cyanidin-3-glucoside, and delphinidin-3-glucoside. On the other hand, a higher content of ascorbic acid and better antioxidant activity was determined in the samples dried at 70 °C. CONCLUSION: According to the results obtained, spreading thickness of 2 mm and drying temperature of 70 °C were suggested for pomegranate juice powder production by foam mat drying. Overall, it was demonstrated that saponin-rich extracts are a good foaming agent alternative that provides foaming at very low concentrations and a product with high quality. © 2020 Society of Chemical Industry.


Subject(s)
Aesculus/chemistry , Fruit and Vegetable Juices/analysis , Fruit/chemistry , Plant Extracts/analysis , Pomegranate/chemistry , Saponaria/chemistry , Saponins/chemistry , Desiccation , Food Additives/chemistry , Powders/chemistry
12.
Arch Virol ; 165(6): 1445-1451, 2020 Jun.
Article in English | MEDLINE | ID: mdl-32248293

ABSTRACT

The use of the nanocapsulated adjuvant Sapomax increased the expression of innate immunity genes (H2Q10, Ddx58, Tyk2, Tlr3, Tlr7, and TNF) responsible for the primary recognition of influenza virus, i.e., those belonging to the RLR and TLR families; genes involved in stimulating the production of type I and III IFN and pro-inflammatory cytokines; and Th1 and Th2 cellular immunity genes (Ccr4, Ccr5, IFNγ, IL-2, IL-4, and IL-10) responsible for triggering regulatory immune mechanisms in the cell. The high immunological activity of the plant-derived nanocapsulated adjuvant Sapomax may be used to enhance the efficacy of vaccines.


Subject(s)
Adjuvants, Immunologic/administration & dosage , Immunity, Innate/drug effects , Saponaria/chemistry , Vaccines/immunology , Adjuvants, Immunologic/genetics , Animals , Cytokines/immunology , Drug Compounding , Female , Male , Mice , Mice, Inbred BALB C , Nanocapsules , Th1 Cells/drug effects , Th1 Cells/immunology , Th2 Cells/drug effects , Th2 Cells/immunology
13.
Mol Ecol ; 27(23): 4947-4959, 2018 12.
Article in English | MEDLINE | ID: mdl-30372557

ABSTRACT

Multiple infections (co-occurrence of multiple pathogen genotypes within an individual host) can have important impacts on diseases. Relatedness among pathogens can affect the likelihood of multiple infections and their consequences through kin selection. Previous studies on the castrating anther-smut fungus Microbotryum lychnidis-dioicae have shown that multiple infections occur in its host plant Silene latifolia. Relatedness was high among fungal genotypes within plants, which could result from competitive exclusion between unrelated fungal genotypes, from population structure or from interactions between plant and fungal genotypes for infection ability. Here, we aimed at disentangling these hypotheses using M. saponariae and its host Saponaria officinalis, both experimentally tractable for these questions. By analysing populations using microsatellite markers, we also found frequent occurrence of multiple infections and high relatedness among strains within host plants. Infections resulting from experimental inoculations in the greenhouse also revealed high relatedness among strains co-infecting host plants, even in clonally replicated plant genotypes, indicating that high relatedness within plants did not result merely from plant x fungus interactions or population structure. Furthermore, hyphal growth in vitro was affected by the presence of a competitor growing nearby and by its genetic similarity, although this latter effect was strain-dependent. Altogether, our results support the hypothesis that relatedness-dependent competitive exclusion occurs in Microbotryum fungi within plants. These microorganisms can thus respond to competitors and to their level of relatedness.


Subject(s)
Basidiomycota/genetics , Basidiomycota/pathogenicity , Plant Diseases/microbiology , Saponaria/microbiology , Flowers/microbiology , Genetic Variation , Genotype , Microsatellite Repeats , Plant Infertility , Saponaria/genetics , Virulence
14.
Ecotoxicol Environ Saf ; 161: 676-682, 2018 10.
Article in English | MEDLINE | ID: mdl-29935432

ABSTRACT

Azole fungicides constitute an extensive group of potential emerging pollutants which can be found in natural environment. This study focuses on the biodegradation of clotrimazole and the characterization of cell surface properties of microorganisms capable of degradation of this compound. The influence of long-term contact of bacteria with clotrimazole and the impact of the addition of Saponaria officinalis extract on cell surface modification was also checked. The biodegradation of clotrimazole did not exceed 70%. The presence of plant extract increased biodegradation of fungicide. The cells metabolic activity after one-month exposure to clotrimazole was the highest for each tested strain. Moreover, metabolic stress led to a strong modification of cell surface properties. The results are promising for determining the impact of clotrimazole on environmental microorganisms.


Subject(s)
Antifungal Agents/metabolism , Clotrimazole/metabolism , Environmental Pollutants/metabolism , Saponins , Bacteria/metabolism , Biodegradation, Environmental , Plant Extracts/chemistry , Saponaria/chemistry , Stress, Physiological , Surface Properties , Surface-Active Agents
15.
Biochim Biophys Acta ; 1854(10 Pt A): 1357-64, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26096917

ABSTRACT

Saporin-S6 is a plant toxin belonging to the type 1 ribosome-inactivating protein (RIP) family. Since it was extracted and isolated from Saponaria officinalis for the first time almost thirty years ago, the protein has been widely studied mainly for its potential applications in anti-tumour and anti-viral infection therapy. Like other RIPs, saporin-S6 is particularly effective in the form of immunotoxin conjugated with monoclonal antibodies and its chemico-physical characteristics made the protein a perfect candidate for the synthesis, development and use of saporin-S6-based chimeric toxins. The high stability of the protein against different denaturing agents has been broadly demonstrated, however, its complete thermal unfolding characterization has not already been performed. In this work we analyse in detail structure, thermostability and unfolding features by means of infrared spectroscopy coupled with two-dimensional correlation spectroscopy. Our data showed that saporin-S6 in solution at neutral pH exhibits a secondary structure analogue to that of the crystal and confirmed its good stability at moderately high temperatures, with a temperature of melting of 58°C. Our results also demonstrated that the thermal unfolding process is non-cooperative and occurs in two steps, and revealed the sequence of the events that take place during the denaturation, showing a higher stability of the N-terminal domain of the protein.


Subject(s)
Ribosome Inactivating Proteins, Type 1/chemistry , Saponaria/chemistry , Seeds/chemistry , Hot Temperature , Hydrogen-Ion Concentration , Models, Molecular , Protein Denaturation , Protein Stability , Protein Structure, Secondary , Protein Structure, Tertiary , Protein Unfolding , Ribosome Inactivating Proteins, Type 1/isolation & purification , Saporins , Solutions , Spectroscopy, Fourier Transform Infrared
16.
Planta Med ; 82(18): 1525-1531, 2016 Dec.
Article in English | MEDLINE | ID: mdl-27392242

ABSTRACT

Triterpenoidal saponins are synthesized in the roots of Saponaria officinalis L. The same plant is also a source for the toxin Saporin, which is a ribosome-inactivating protein. Triterpenoidal saponins are known to increase the cytotoxicity of Saporin by modulating its intracellular trafficking. Here, we investigated if the combinatorial effects elicited by purified saponins and Saporin can be applied to increase the therapeutic efficacy of the immunotoxin Saporin-Rituximab. First, saponins were purified by high-performance liquid chromatography. Thereafter, their intrinsic cytotoxicity was evaluated on Ramos cells with no observed effect up to 5 µg/mL, however, saponins increased the cytotoxicity of Saporin, while no influence was observed on its N-glycosidase activity. Saporin-Rituximab bound to CD20 in Ramos cells and, in the absence of saponins, had a GI50 (concentration inhibiting cell growth to 50 %) of 7 nM. However, in the presence of a nontoxic concentration of saponins, the GI50 of Saporin-Rituximab was 0.01 nM, a nearly 700-fold increase in efficacy. Moreover, two further immunotoxins, namely Saporin-anti-CD22 and Saporin-anti-CD25, were tested in combination with saponins yielding enhancement factors of 170-fold and 25-fold, respectively. All three receptors are present in Ramos cells and the differences in cytotoxicity enhancement may be explained by the differing expression levels of the cellular receptors. The application of purified saponins from S. officinalis L. is therefore a new strategy to potentially improve the cytotoxicity and therapeutic efficacy of Rituximab-immunotoxins for the treatment of B-cell lymphoma.


Subject(s)
Immunotoxins/pharmacology , Lymphoma, B-Cell/pathology , Ribosome Inactivating Proteins, Type 1/pharmacology , Rituximab/pharmacology , Saponaria/chemistry , Saponins/pharmacology , Cell Line , Cell Survival/drug effects , Chromatography, High Pressure Liquid , Drug Synergism , Humans , Immunotoxins/chemistry , Immunotoxins/isolation & purification , Ribosome Inactivating Proteins, Type 1/chemistry , Ribosome Inactivating Proteins, Type 1/isolation & purification , Saponins/chemistry , Saponins/isolation & purification , Saporins
17.
Biochemistry ; 54(29): 4565-74, 2015 Jul 28.
Article in English | MEDLINE | ID: mdl-26091305

ABSTRACT

Saporin L3 from Saponaria officinalis (soapwort) leaves is a type 1 ribosome-inactivating protein. It catalyzes the hydrolysis of oligonucleotide adenylate N-ribosidic bonds to release adenine from rRNA. Depurination sites include both adenines in the GAGA tetraloop of short sarcin-ricin stem-loops and multiple adenines within eukaryotic rRNA, tRNAs, and mRNAs. Multiple Escherichia coli vector designs for saporin L3 expression were attempted but demonstrated high toxicity even during plasmid maintenance and selection in E. coli nonexpression strains. Saporin L3 is >10(3) times more efficient at RNA deadenylation on short GAGA stem-loops than saporin S6, the saporin isoform currently used in immunotoxin clinical trials. We engineered a construct for the His-tagged saporin L3 to test for expression in Pichia pastoris when it is linked to the protein export system for the yeast α-mating factor. DNA encoding saporin L3 was cloned into a pPICZαB expression vector and expressed in P. pastoris under the alcohol dehydrogenase AOX1 promoter. A fusion protein of saporin L3 containing the pre-pro-sequence of the α-mating factor, the c-myc epitope, and the His tag was excreted from the P. pastoris cells and isolated from the culture medium. Autoprocessing of the α-mating factor yielded truncated saporin L3 (amino acids 22-280), the c-myc epitope, and the His tag expressed optimally as a 32 kDa construct following methanol induction. Saporin L3 was also expressed with specific alanines and/or serines mutated to cysteine. Native and Cys mutant saporins are kinetically similar. The recombinant expression of saporin L3 and its mutants permits the production and investigation of this high-activity ribosome-inactivating protein.


Subject(s)
Plant Proteins/chemistry , Ribosome Inactivating Proteins, Type 1/chemistry , Saponaria/enzymology , Amino Acid Substitution , Base Sequence , Catalytic Domain , Hydrogen-Ion Concentration , Kinetics , Mutagenesis, Site-Directed , Pichia , Plant Proteins/biosynthesis , Plant Proteins/genetics , RNA/chemistry , RNA Cleavage , Ribosome Inactivating Proteins, Type 1/biosynthesis , Ribosome Inactivating Proteins, Type 1/genetics , Saporins , Substrate Specificity
18.
J Biol Chem ; 288(18): 12500-10, 2013 May 03.
Article in English | MEDLINE | ID: mdl-23486480

ABSTRACT

Caryophyllaceae-type cyclic peptides (CPs) of 5-12 proteinogenic amino acids occur in 10 plant families. In Saponaria vaccaria (Caryophyllaceae), they have been shown to be formed from linear peptide precursors derived from ribosomal translation. There is also evidence for such precursors in other members of the Caryophyllaceae, Rutaceae, and Linaceae families. The biosynthesis of CP in the developing seeds of S. vaccaria was investigated with respect to the enzymes involved in precursor processing. Through biochemical assays with seed extracts and synthetic peptides, an enzyme named oligopeptidase 1 (OLP1) was found that catalyzes the cleavage of intermediates at the N terminus of the incipient CP. A second enzyme, peptide cyclase 1 (PCY1), which was separated chromatographically from OLP1, was found to act on the product of OLP1, giving rise to a cyclic peptide and concomitant removal of a C-terminal flanking sequence. PCY1 was partially purified, and using the methods of proteomics, a full-length cDNA clone encoding an enzyme matching the properties of PCY1 was obtained. The substrate specificity of purified recombinant PCY1, believed to be the first cloned plant enzyme whose function is peptide cyclization, was tested with synthetic peptides. The results are discussed in the light of CP biosynthetic systems of other organisms.


Subject(s)
Peptide Biosynthesis, Nucleic Acid-Independent/physiology , Peptides, Cyclic/biosynthesis , Plant Proteins/metabolism , Saponaria/enzymology , Seeds/enzymology , Serine Proteases/metabolism , Amino Acid Sequence , Cloning, Molecular , DNA, Complementary/genetics , Molecular Sequence Data , Peptides, Cyclic/genetics , Plant Proteins/genetics , Saponaria/genetics , Seeds/genetics , Serine Proteases/genetics
19.
Biosci Biotechnol Biochem ; 78(2): 288-95, 2014.
Article in English | MEDLINE | ID: mdl-25036683

ABSTRACT

Saponins have the potential to favorably modulate rumen fermentation, but there is generally a lack of the chemical structures associated with the described effects. The activity of extracts from Calendula officinalis and Saponaria officinalis in the rumen was evaluated in vitro. The S. officinalis root extract, reduced CH4 production by 8.5% and increased total VFA concentration by 25.2%. C. officinalis and S. officinalis root extracts and the S. officinalis aerial part extract decreased the acetate to propionate ratio from 8.6 to 17.4%, according to the extract. An HPLC-ELSD analysis indicated that the saponin content ranged from 43.6 to 57.6 mg/g of dry matter (DM) in the C. officinalis extracts and from 224.0 to 693.8 mg/g of DM in the S. officinalis extracts, expressed as the hederacoside C equivalent. Identification of the saponin compounds present in the extracts by HPLC-MS(n) suggested that the saponin profile modulated the biological activities, showing the importance of determining the structure of saponins when evaluating extracts.


Subject(s)
Calendula/chemistry , Fermentation/drug effects , Plant Extracts/pharmacology , Rumen/drug effects , Rumen/metabolism , Saponaria/chemistry , Saponins/metabolism , Animals
20.
Bull Exp Biol Med ; 156(3): 366-9, 2014 Jan.
Article in English | MEDLINE | ID: mdl-24771377

ABSTRACT

The effects of triterpene glycosides (saponins) extracted from Saponaria officinalis L. radices, on the cellular and humoral innate immunity factors were studied. Saponins stimulated the phagocytic, bactericidal, and adhesion activities of polymorphonuclear leukocytes. Optimal conditions of saponin treatment (dose and duration) were determined for mice. Saponins promoted the maturation of human peripheral blood dendritic cells, which was proven by high expression of CD83 (terminal differentiation marker) and CD86 (bone-stimulating molecule) and of HLA-DR and HLA-ABC molecules on the cell membrane. Saponins modulated the production of TNF-α, IL-1ß, IL-4, IL-6, and IFN-γ in cultured peripheral blood intact cells. The results help to understand some mechanisms of the effects of saponins extracted from Saponaria officinalis L. radix on the cellular and humoral factors of innate immunity and demonstrate good prospects of their practical use.


Subject(s)
Immunity, Innate/drug effects , Plant Extracts/pharmacology , Saponaria/drug effects , Triterpenes/pharmacology , Animals , Cells, Cultured , Glycosides/chemistry , Humans , Mice , Triterpenes/chemistry
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