The aim of the study was to investigate the effect of cigarette smoke on the production and characterization of exopolysaccharides (EPSs) produced by Bifidobacterium. Cigarettes of Shanhua brand (nicotine: 1.1 mg, tar: 11 mg) were utilized to prepare a cigarette smoke condensate (CSC). The standard strain of Bifidobacterium animalis was cultured in MRS media under anaerobic addition of CSC. The results showed that CSC significantly decreased the growth of B. animalis as well as EPSs and acetic acid production. Furthermore, two EPSs fractions (Fr-I and Fr-II) were isolated and purified for chemical and molecular determination. By comparison with control, CSC was found to be of great impact on EPSs carbohydrate composition. The molecular weight mass of Fr-I changed from 3.33×10 5 g/mol (without CSC) to 2.99×10 5 (with CSC). In conclusion, in vitro studies revealed that CSC was directly able to affect the production of metabolites for B. animalis, which could be an essential factor in certain pathological disorders.
SummaryIn this study, the eff ects of the addition of Tween 80 and acetone on secretion, structure and antioxidant activities of Lentinus tigrinus exopolysaccharides (EPS) were investigated. It was found that Tween 80 and acetone displayed a stimulatory eff ect on EPS secretion. The EPS obtained by the addition of Tween 80 (EPS-T), acetone (EPS-A) and control (EPS--C) were purifi ed by Sepharose CL-6B gel fi ltration chromatography and molecular mass of purifi ed fractions was estimated to be 22.1, 137 and 12 kDa, respectively. Monosaccharide composition analysis indicated that EPS-T, EPS-A and EPS-C were mainly composed of glucose and mannose. Congo Red test indicated that EPS-T and EPS-A had a highly ordered conformation of triple helix, while EPS-C had a random coil conformation. Furthermore, EPS-A exhibited higher DPPH scavenging and antiproliferative activities than EPS--C and EPS-T, which might be att ributed to the molecular mass.
A soil isolate, Penicillium janthinellum sw09 has been found to produce significant amounts of an extracellular pectinase subsequently characterized as exo-polygalacturonase (exo-PG). By optimizing growth conditions, P. janthinellum sw09 produced high amount of exo-PG (16.54 units/mL). The crude enzyme was purified by gel filtration chromatography and two exo-PG activity peaks (designated as PGI and PGII) were revealed. On SDS-PAGE analysis, purified PGII using DEAE-Sepharose FF column, was found to be a single band with a molecular mass of 66.2 kDa. The purified PGII exhibited maximal activity at the temperature of 45 o C and pH 5.0. The stability profiles show that PGII is more stable in the pH range of 4.0-8.0 and below 60 o C. The K m and V max for the enzyme was 1.74 mg/mL and 18.08 μmol/ (mL•min), respectively. Due to this enzymatic characterization, this pectinase is an attractive candidate for applications in degradation of pectin.
Background:Many exopolysaccharides from the endophytes in medicinal plants possess various potential bioactivities.Materials and Methods:The endophytic fungus JY25 was isolated from the leave of the Chinese medicinal plant Gynostemma pentaphylla and identified as Chaetomium sp. by its phylogenetic and physiological analysis. One exopolysaccharide (EPS) fraction was isolated from the fermentation broth by ethanol precipitation and purified by gel filtration chromatography on Sepharose CL-6B. The molecular characteristics were examined by GC-MS, FT-IR, and multiangle laser light scattering (MALLS).Results:The monosaccharide composition analysis indicated that the purified EPS was mainly composed of glucose, mannose, arabinose, and galactose with the molecular ratio of 78.29: 8.99: 8.64: 4.08. FT-IR spectral analysis of the purified EPS revealed prominent characteristic groups, such as carbonyl bond, pyranose ring, and so on. The weight-average molar mass and the polydispersity ratio of the EPS were revealed to be 1.961×104 g/mol and 1.838, respectively. Furthermore, thermo gravimetric analysis (TGA) indicated that the degradation temperature of the purified EPS was 305° C. The purified EPS from the endophytic fungus Chaetomium sp. displayed antioxidant and antiproliferative activities.Conclusion:The results demonstrated that the EPS could be used as a healthful food and material source in pharmaceutical industries.SUMMARY An exopolysaccharides (EPS) with antioxidant and antiproliferative activities from an endophytic fungus Chaetomium sp. was reported. Abbreviations used: ANOVA: Analysis of variance; DPPH: 2,2-diphenyl-1-picrylhydrazyl; EPS: Exopolysaccharide; FT-IR: Fourier transform infrared spectroscopy; GC-MS: Gas chromatography–mass spectrometry; Mw: Mass weight; MALLS: Multiangle laser light scattering; SEC: Size Exclusion Chomatography; SPSS: Statistical Package of the Social Science; TGA: Thermo gravimetric analysis; TFA: Trifluoroacetic acid.
ObjectiveTriple-negative breast cancer (TNBC) is distinguished by early recurrence and metastases, a high proclivity for treatment resistance, and a lack of targeted medicines, highlighting the importance of developing innovative therapeutic techniques. Salvia chinensis Benth (SCH) has been widely studied for its anticancer properties in a variety of cancers. However, its significance in TNBC treatment is rarely discussed. Our study investigated the anticancer effect of SCH on TNBC and the underlying mechanisms.MethodsFirst, we used clonogenic, cell viability, flow cytometry, and Transwell assays to assess the effect of SCH on TNBC. Bioinformatic studies, especially network pharmacology-based analysis and RNA sequencing analysis, were performed to investigate the constituents of SCH and its molecular mechanisms in the suppression of TNBC. High-performance liquid chromatography and thin-layer chromatography were used to identify two major components, quercetin and β-sitosterol. Then, we discovered the synergistic cytotoxicity of quercetin and β-sitosterol and assessed their synergistic prevention of cell migration and invasion. Breast cancer xenografts were also created using MDA-MB-231 cells to test the synergistic therapeutic impact of quercetin and β-sitosterol on TNBC in vivo. The impact on the DNA damage and repair pathways was investigated using the comet assay and Western blot analysis.ResultsOur findings showed that SCH decreased TNBC cell growth, migration, and invasion while also inducing cell death. We identified quercetin and β-sitosterol as the core active components of SCH based on a network pharmacology study. According to RNA sequencing research, the p53 signaling pathway is also regarded as a critical biological mechanism of SCH treatment. The comet assay consistently showed that SCH significantly increased DNA damage in TNBC cells. Our in vivo and in vitro data revealed that the combination of quercetin and β-sitosterol induced synergistic cytotoxicity and DNA damage in TNBC cells. In particular, SCH particularly blocked the inter-strand cross-link repair mechanism and the double-strand breach repair caused by the homologous recombination pathway, in addition to inducing DNA damage. Treatment with quercetin and β-sitosterol produced similar outcomes.ConclusionThe current study provides novel insight into the previously unknown therapeutic potential of SCH as a DNA-damaging agent in TNBC.
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