Gleditsia Saponin Research Articles

A macromolecular Gleditsia Saponin induces apoptosis of leukemia cells through Fas-mediated death receptor pathway and mitochondrial pathway

Gleditsia sinensis Lam., a natural product derived from medicine and food, exhibits outstanding biological activity, with its primary active component being triterpenoid saponin. In this study, a new macromolecular Gleditsia Saponin (GS) with a molecular weight of 27.562 kDa was extracted from Gleditsia sinensis Lam.. The study focused on examining the impact of GS on the suppression of cell growth and the initiation of apoptosis in leukemia K562 and L1210 cells. The results showed that GS notably suppressed the growth of leukemia cells in a manner reliant on concentration. Morphological and Annexin V-FITC/PI staining analyses revealed characteristic features of apoptosis in leukemia cells treated with GS. Moreover, cell cycle analysis indicated that GS effectively arrested leukemia cell progression at the G0/G1 phase. Mitochondrial membrane potential (ΔΨm) was disrupted, and abnormal accumulation of reactive oxygen species was also detected. In addition, GS could induce cell apoptosis through the Fas-mediated death receptor pathway and mitochondrial pathway, leading to the blocking of leukemia cells in the G1 phase. In summary, these results provide theoretical backing and fresh perspectives for the development and utilization of GS as a functional health food.

Evaluation of interfacial, micellar, and foaming properties of the solutions comprising fluorocarbon surfactant, cocamidopropyl betaine, and Gleditsia saponin as fire-extinguishing agents

The interfacial, micellar, and foam properties of unitary, binary, and ternary aqueous solutions containing a non-ionic fluorocarbon surfactant (FC-170C), an ionic hydrocarbon surfactant (CAB), and Gleditsia saponin (GS) were comprehensively investigated. FC-170C showed the highest surface activity and micellization ability, CAB and GS showed the strongest foaming ability and foam stability, respectively. For binary mixtures, the interaction in each pair of mixtures synergistically enhanced the ability of surface tension reduction, but had little synergistic effect on the efficiency of surface tension reduction. For the ternary mixtures, at an optimal formulation of 7.43 × 10-6 mol/L FC-170C, 5.26 × 10-3 mol/L CAB, and 1.60 × 10-4 mol/L GS, the values of γ, h0, R5min, and bubble size were 17.3 mN/m, 117 mm, 98%, and 115–116 μm, respectively. In particular, the required amount of fluorocarbon was reduced to 6.78 mg/kg, which is significantly<50 mg/kg limit set by international conventions for fire extinguishing agents.

Effective lactic acid production from waste paper using Streptococcus thermophilus at low enzyme loading assisted by Gleditsia saponin

Waste paper has considerable potential as a raw material for lactic acid (LA) production due to high cellulose content, abundance and low cost. In this study, four kinds of waste papers were used for LA production through simultaneous saccharification and fermentation (SSF) by Streptococcus thermophilus. The SSF of office paper achieved the highest LA concentration (39.71 g/L), while the highest LA yield was observed for magazine (99.56%), followed by office paper (82.85%). High LA concentration is unfavorable to total LA conversion because of product inhibition. However, the addition of Gleditsia saponin (GS) could obtain both high yield and high concentration of LA at a low enzyme loading, indicating that product inhibition could be moderated. A lactic acid yield of 86.30% was obtained from office paper at an enzyme loading of 9 FPU/g-cellulose with GS, which was higher than that of without GS at a higher loading of 18 FPU/g-cellulose.

Gleditsia Saponin C Induces A549 Cell Apoptosis via Caspase-Dependent Cascade and Suppresses Tumor Growth on Xenografts Tumor Animal Model.

Saponins are natural compounds and possess the most promising anti-cancer function. Here, a saponin gleditsia saponin C (GSC), extracted from gleditsiae fructus abnormalis, could induce apoptosis of lung tumor cell line A549 via caspase dependent cascade and this effect could be prevented by the caspase inhibitors. In addition, GSC induced cell death companied with an increase ratio of Bax:Bcl-2 and inhibition of ERK and Akt signaling pathways. Meanwhile, GSC suppressed TNFα inducing NF-κB activation and increased the susceptibility of lung cancer cell to TNFα induced apoptosis. Furthermore, on mouse xenograft model, GSC significantly suppressed tumor growth and induced cancer cell apoptosis, which validated the anti-tumor effect of GSC. Based on these results, GSC might be a promising drug candidate of anti-lung cancer for its potential clinical applications.

Enhancement of high-solids enzymatic hydrolysis and fermentation of furfural residues by addition of Gleditsia saponin

Gleditsia saponin (GS) assisted high-solids enzymatic hydrolysis and fermentation for alkaline peroxide (AP)-pretreated furfural residues (FRs) was investigated. The addition of GS increased the glucose yield of untreated FRs to 74.8% in high-solids-loading enzymatic hydrolysis. AP pretreatment before enzymatic hydrolysis resulted in a high sugar concentration (115.4g/L) and a glucose yield of 57.5% by the end of hydrolysis at 20% solids loading. It was suggested that GS could improve significantly the glucose yields of FRs with high lignin content. After simultaneous saccharification and fermentation (SSF), the total cellulose conversion of FRs with 14.3% lignin content reached 82.2% and the highest ethanol concentration (69.1g/L) was achieved, which was 121% higher than that of untreated FRs (30.1g/L). The results indicated that the main reason behind the decreased glucose yield at high-solids loadings was product inhibition. Moreover, the lignin content plays an important role in the enhancement of GS-assisted high-solids enzymatic hydrolysis of AP-treated FRs.

Enhanced Ethanol Production with Mixed Lignocellulosic Substrates from Commercial Furfural and Cassava Residues

Simultaneous saccharification and fermentation (SSF) is an attractive process configuration for bio-ethanol production. Further reductions in process cost of SSF are expected with the use of waste agricultural or industrial materials as feedstock. In the current study, two industrial lignocellulosic wastes, cassava residues (CR) and furfural residues (FR), were combined during SSF for ethanol production due to their value-added applications and positive environmental impacts. After CR were liquefied and saccharified, saccharification liquid was added to SSF of FR. The effect of substrate fractions was investigated in terms of ethanol yield, byproduct concentration and the number of yeast cells. Besides, a natural surfactant, Gleditsia saponin, was added to investigate the effect of FR lignin on SSF with 20% substrate concentration. The results showed that increasing the ratio of CR/FR improved the ethanol yield and that the ethanol yield was also increased gradually by increasing the substrate concentration from 6% to 12%. A high ethanol concentration of 36.0 g/L was obtained under the condition of CR:FR = 2:1 with 12% substrate concentration, reaching 71.1% of the theoretical yield. However, Gleditsia saponin did not affect the ethanol yield, indicating the insignificant effect of lignin in SSF with low lignin content in the reaction system.

Effects of Gleditsia saponin on high-solids enzymatic hydrolysis of furfural residues

Surfactants are popular additives for the prevention of the inhibitory effect of lignin on cellulolytic enzymes. Saponin is a type of natural surfactant extracted from Gleditsia spp. In this study, the effect of saponin on the high-solids-loading enzymatic hydrolysis of furfural residues (FRs) with a lignin content of 45% was investigated. The optimal dosage of Gleditsia saponin (GS) was determined to be 25-fold greater than its critical micelle concentration (CMC; 0.16g/L). The addition of GS increased the cellulose conversion by up to 52.63% in high-solids-loading enzymatic hydrolysis. Due to the addition of GS, a maximum cellulose conversion of 74.88% was achieved after FRs had hydrolyzed for 120h at a solids loading of 20% (w/w) with 30 filter paper unit (FPU)/g-cellulose. The decrease in cellulose conversion was found to exhibit a linear correlation with an increase in the solids loadings from 20.0% to 27.5% (w/w). The surface structural variations of hydrolytic residues were characterized by X-ray photoelectron spectroscopy (XPS). The results indicated that the carbonyl groups of lignin facilitated the binding of GS to the surface of lignin, which resulted in a decrease in the adsorption of between the enzyme and lignin. Interestingly, GS could maintain enzymatic activity in the supernatants.

Bioactive triterpenoid saponins and phenolic compounds against glioma cells

A total of 54 natural origin compounds were evaluated for their activity in inhibiting the proliferation of glioma cells. Results showed that four Aesculus polyhydroxylated triterpenoid saponins (3–6), six Gleditsia triterpenoid saponins (7–12), and five phenolic compounds (43–46, 51) had dose-dependent activity suppressing the proliferation of both C6 and U251 cells. Structure–activity relationship analysis suggested that the acetyl group at C-28 for the Aesculus saponins and the monoterpenic acid moiety for the Gleditsia saponins could be critical for the activity of these active compounds. Aesculioside H (4), gleditsioside A (7), and feuric acid 3,4-dihydroxyphenethyl ester (FADPE, 46) were the three most active compounds from the different types of the active compounds and induced apoptosis and necrosis in glioma cells.

Caspicaosides E–K, triterpenoid saponins and cytotoxic acylated saponins from fruits of Gleditsia caspica Desf.

Seven bisdesmosidic triterpenoid saponins named caspicaosides E–K, were isolated from the methanolic fruit extract of Gleditsia caspica Desf. Their structures were determined by 1D and 2D NMR spectroscopy as well as high resolution mass spectrometry and acid hydrolysis. The saponins comprised echinocystic acid or oleanolic acid as the aglycone and saccharide moieties at C-3 and C-28. Like most Gleditsia saponins, the oligosaccharide moiety at C-3 was identified as β-d-xylopyranosyl-(1→2)-α-l-arabinopyranosyl-(1→6)-β-d-glucopyranosyl. The common oligosaccharide moiety linked to C-28 was determined as β-d-xylopyranosyl-(1→3)-β-d-xylopyranosyl-(1→4)-α-l-rhamnopyranosyl-(1→2)-β-d-glucopyranosyl with the presence of additional β-d-galactopyranose unit and/or another α-l-rhamnopyranose moiety. The C-28 saccharide moiety was acylated with a monoterpenic acid unit or a monoterpenic acid linked to a monoterpene-arabinoside unit at the ester-β-d-glucopyranose C-6. Esterification of C-2 and C-3 hydroxyl groups of the terminal α-l-rhamnopyranose unit with a monoterpenic acid and epoxy-monoterpenic acid units, respectively, or with two identical monoterpenic acid units, was also shown. The acylated saponins caspicaosides G–K were assayed for their in vitro cytotoxicities against the three cell lines HCT116, HepG2 and MCF7. The tested saponins showed moderate to strong activities.

Quantitive analysis of gleditsia saponins in the fruits of Gleditsia sinensis Lam. by high performance liquid chromatography

A high performance liquid chromatography method was developed for simultaneous quantification of 29 ingredients occurring as a very complex mixture in the gleditsia fruits. The analysis was performed on an ODS-2 Hypersil column (250mm×4.6mm, 5μm) with a binary gradient mobile phase of water and acetonitrile both containing 0.1% acetic acid. The method was validated in terms of linearity, sensitivity, stability, precision, and accuracy. It was found that this method had linearity with R2 at 0.99889–0.99997 in the test range of 1.0–24.0μg. The limit of detection (LOD) and limit of quantification (LOQ) for 16 tested reference saponins were 0.24–0.39μg and 1.0–1.2μg, respectively. The relative standard deviations (RSDs %) for intra-day and inter-day repeatability were not more than 3.11% and 4.02%, respectively. The analyzed samples were stable for at least 48h. The spike recoveries for eight analyzed saponins were 99.68–102.17%. The established HPLC analytic method was successfully used to determine the concentrations of 29 compounds including 19 gleditsia saponins and ten unidentified ingredients in eight commercial gleditsia fruits from different sources. The results from this study suggested that this newly developed HPLC method could be used for qualitative and quantitative analysis of the saponins in the gleditsia fruits and the gleditsia extracts that used for animal study and other purpose.

The Effect of &lt;i&gt;Gleditsia&lt;/i&gt; Saponin on Simultaneous Saccharification and Fermentation of Furfural Residue for Ethanol Production

Simultaneous saccharification and fermentation is an attractive process configuration for bioethanol. However, the process cost in SSF is still high. Further reductions in cost are expected with reduction of enzyme addition and the use of waste agricultural or industrial materials as feedstock. Surfactant addition is a simple method to reduce the amount of enzyme loading. In the present work we explore the effect of Gleditsia Saponin, an inexpensive and natural surfactant, on SSF of furfural residue, a main waste of furfural industry in China, for ethanol production. Gleditsia Saponin addition even at low concentration, 0.04 g/l, increased the ethanol yield by 9.9%. Meanwhile, when Gleditsia Saponin addition was 0.16 g/l, the yield was increased by 22.8% and the enzyme activity significantly increased in the liquid fraction at the end of SSF, which would contribute to recover and recycle enzyme. Gleditsia Saponin addition at 1.25 g/l had a better performance in improving SSF efficiency than that of 2.5 g/l Tween-20 addition. The positive effects of Gleditsia Saponin addition on SSF, such as less enzyme loading and shorter residence time could also been observed.

Surface Properties of Gleditsia Saponin and Synergisms of Its Binary System

A gleditsia saponin (GS) is isolated and purified from the fruits of Gleditsia sinensis Lam. Its surface and thermodynamic properties in aqueous solution at 0, 15, 25, 35, 45, and 60°C have been investigated by surface tension measurement. The major data at 25°C are the critical micelle concentration (cmc), 1.10E‐3 mol · L−1; the surface tension at cmc (γ cmc ), 34.4 mN · m−1; the maximum surface excess concentration (Γmax), 2.33E‐6 mol · m2; the minimum area per molecule (A cmc ), 0.712 nm2; the standard adsorption energy (ΔG ad 0), −32.9 kJ · mol−1; and the standard micellization energy (ΔG m 0), −16.9 kJ · mol−1. The molecular interaction parameters, β s and β m , for the binary systems of GS with a cationic surfactant (C16H33NMe3Br), an anionic surfactant (C12H25SO3Na), and a nonionic surfactant (C8H17Ph(EO)10OH) have been determined. Based on the regular solution treatments of Rubingh, Rosen, and their coworkers, the conditions of synergism in surface tension reduction effectiveness have been extended. The condition to produce this synergism and the corresponding parameters at the maximum synergism point, i.e., the mole fractions of surfactant 1 in the solution phase (α1*), the critical micelle concentration of mixtures (cmc 12*), and the surface tension at this concentration (γ cmc12*), are given. The results show that of the three binary systems, synergism in micelle formation can occur only in the mixture of GS with cationic surfactants; synergism in surface tension reduction efficiency can occur in the mixture of GS with both cationic surfactant and nonionic surfactants, but, all the binary systems of GS with ionic and nonionic surfactants can show remarkable synergism in surface tension reduction effectiveness.

Structure-activity relationships of saponins from Gleditsia sinensis in cytotoxicity and induction of apoptosis.

Thirteen compounds were isolated from the anomalous fruits of Gleditsia sinensis on the basis of bioassay-guided fractionation. These saponins together with six analogues or related compounds were tested for their cytotoxicities against six tumor cell lines by the MTT method. The induction of apoptosis in HL-60 cells by these compounds was determined through flow cytometric analysis. Some structure-activity relationships in cytotoxicity and induction of apoptosis were identified. The evaluation of the cytotoxicity and the ability to induce apoptosis revealed that some important structural features are required for activity. A valuable model which enables prediction of their activities was established and may be employed for the drug design of new Gleditsia saponin analogues.

Triterpenoidal saponins from Gleditsia sinensis.

Six bisdesmosidic triterpenoidal saponins, gleditsiosides H-K and gleditsia saponins C' and E', were isolated from the anomalous fruits of Gleditsia sinensis. Their structures were established by a combination of extensive NMR (DEPT, DQF-COSY, HETCOR, HOHAHA, HMBC and ROESY) studies and chemical degradation.

Anti-AIDS agents, 21. Triterpenoid saponins as anti-HIV principles from fruits of Gleditsia japonica and Gymnocladus chinensis, and a structure-activity correlation.

Gleditsia saponin C [1] and gymnocladus saponin G [2] were isolated from Gleditsia japonica and Gymnocladus chinensis, respectively, as anti-HIV principles. Compounds 1 and 2 demonstrated inhibitory effects against HIV replication in H-9 cells with EC50 values of 1.1 and 2.7 microM, respectively. Evaluation of the anti-HIV activities of the prosapogenins of 1 and 2 suggested that the unusual monoterpenyl moieties in 1 and 2 are essential for their anti-HIV activity. Derivatives of echinocystic acid [8], the aglycone of compound 1, were also prepared and evaluated for inhibitory activity against HIV replication. 3,16-Di-O-acetylechinocystic acid [12] was shown to be an anti-HIV agent with an EC50 value of 2.3 microM.

Alteration of Na + permeability in human erythrocytes as studied by 23Na-NMR and inhibition of the kidney Na +,K +-ATPase activities with saponins: Interaction of Gleditsia saponins with human erythrocyte membranes

Interaction of Gleditsia saponin C 1, bis-monoterpenyl echinocystic acid 3,28-O-bisglycoside, with human erythrocyte membranes and artificial membranes was investigated by means of the hemolytic activities, alteration of Na + permeability by 23Na-NMR spectroscopy and inhibition of Na +,K +-ATPase activities. Interaction of Gleditsia saponin C ( 1), bis-monoterpenyl echinocystic acid 3,28-O-bisglycoside, with human erythrocyte membranes and artificial membranes was investigated by means of the hemolytic activities, alteration of Na + permeability by 23Na-NMR spectroscopy and inhibition of Na +,K +-ATPase activities.

Gleditsia Saponins

The chemical structure of gleditsia saponin C (GS-C), a major triterpenoid saponin of the fruit capsule of Gleditsia japonica Miq. (Leguminosae), comprised echinocystic acid, eight sugar units and two monoterpenyl groups ((+)2,6-dimethyl- and (+) 2-hydroxymethyl-6-methyl-6-(S)hydroxy-2- trans-2,7-octadienoyl groups). Methylation analysis of GS-C followed by determination with gas-liquid chromatography and mass spectrometry showed that the monoterpenyl moieties attach to the 2 and 3-positions of terminal L-rhamnose of the C-28 oligoside portion.

Legume saponins of Gleditsia japonica Miquel. IV. 13C-Nuclear magnetic resonance spectral studies for structure elucidation of gleditsia saponins B and C.

Two triterpenoid saponins, gleditsia saponins B (GS-B, XIX) and C (GS-C, XX) which were isolated from Gleditsia japonica cv. 'Saponifera', were characterized as 3, 28-O-bisglycosides of echinocystic acid acylated with monoterpene carboxylic acids. On the basis of 13C-nuclear magnetic resonance spectra and the results of chemical and enzymatic hydrolysis, it was established that the positions of acylation with the mono-terpenes (IV) and (V) were C-2 and C-3, respectively, of the terminal rhamnose of the 28-O-glycoside moiety in the molecules of GS-C and GS-B.

Legume saponins of Gleditsia japonica Miquel. V. 13C Nuclear magnetic resonance spectral studies on the structures of gleditsia saponins D2, G and I.

Three triterpenoid saponins named gleditsia saponins D2 (VII), G (IV) and I (VIII) were isolated from the legume of Gleditsia japonica cv.'Saponifera.' They were characterized as echinocystic acid 3, 28-O-bisglycosides acylated with two monoterpene carboxylic acids IX and X. The locations of these monoterpenes were established by 13C nuclear magnetic resonance spectroscopy.

Legume saponins of Gleditsia japonica Miquel. III. Further desmonoterpenyl glycosides of echinocystic acid.

Two triterpenoid saponins, gleditsia saponins E (GS-E) and G (GS-G), were isolated from legumes of Gleditsia japonica cv. 'Saponifera' (Leguminosae). These saponins contain monoterpene ester moieties. The desmonoterpenyl compounds, GS-E' (C69H112O34) and GS-G' (C64H104O30), were obtained from them by alkaline hydrolysis with K2CO3 and both were identified as echinocystic acid 3, 28-O-bisdesmoside on the basis of physical data and degradation products.