Background: Oligosaccharides are composed of a variable number of monosaccharide units and very important in the biologically diverse of biological systems. Materials and Methods: Crude water-soluble oligosaccharide was extracted from the fruiting bodies with water and then successively purified by DEAE-cellulose 52 and Sephadex G-100 column chromatography, yielding one major oligosaccharides fractions: LES-A. Structural features of Lactarius deliciosus (L. ex Fr.) Gray oligosaccharide (LDGO-A) were investigated by a combination of monosaccharide component analysis by thin layer chromatography, infrared spectra, nuclear magnetic resonance spectroscopy, scanning electron microscopy, and high-performance gel permeation chromatography analysis. Result: The results indicated that LDGO-A was composed of D-glucose and D-xylose, and the average molecular sizes was approximately 945 Da. The anti-tumor activity of LDGO-A was evaluated in vivo. The inhibitory rate in mice treated with 40 mg/kg LDGO-A can reach 40.02%, being the highest in the three doses, which may be comparable to mannatide. Histology of immune organs shows that the tissues arranged more regular and firmer, but the tumor tissue arranged looser in LDGO-A group than those in the control group. Meanwhile, there is no obvious damage to other organs, such as heart. The anti-tumor activity of the LDGO-A was usually believed to be a consequence of the stimulation of the cell-mediated immune response because it can significantly promote the lymphocyte and macrophage cells in the dose range of 100-400 μg/mL in vitro. LDGO-A also effected the expression of some housekeeping genes mRNA in S180 tumor. Conclusion: Accordingly, the LDGO-A might serve as an effective healthcare food and source of natural anti-tumor compounds.
RPL23A gene encodes a ribosomal protein that is a component of the 60S subunit. The protein belongs to the L23P family of ribosomal proteins, which is located in the cytoplasm. The purpose of this paper was to explore the structure and anti-cancer function of ribosomal protein L23A (RPL23A) gene of the Giant Panda (Ailuropoda melanoleuca). The cDNA of RPL23A was cloned successfully from the Giant Panda using RT-PCR technology. We constructed a recombinant expression vector containing RPL23A cDNA and over-expressed it in Escherichia coli using pET28a plasmids. The expression product obtained was purified by using Ni chelating affinity chromatography. Recombinant protein of RPL23A obtained from the experiment acted on Hep-2 cells and human HepG-2 cells, then the growth inhibitory effect of these cells was observed by MTT (3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2H-tetrazolium bromide) assay. The result indicated that the length of the fragment cloned is 506 bp, and it contains an open-reading frame (ORF) of 471 bp encoding 156 amino acids. Primary structure analysis revealed that the molecular weight of the putative RPL23A protein is 17.719 kDa with a theoretical pI 11.16. The molecular weight of the recombinant protein RPL23A is 21.265 kDa with a theoretical pI 10.57. The RPL23A gene can be really expressed in E. coli and the RPL23A protein, fusioned with the N-terminally His-tagged protein, gave rise to the accumulation of an expected 22 KDa polypeptide. The data showed that the recombinant protein RPL23A had a time- and dose-dependency on the cell growth inhibition rate. The data also indicated that the effect at low concentrations was better than at high concentrations on Hep-2 cells, and that the concentration of 0.185 μg/mL had the best rate of growth inhibition of 36.31%. All results of the experiment revealed that the recombinant protein RPL23A exhibited anti-cancer function on the Hep-2 cells. The study provides a scientific basis and aids orientation for the research and development of cancer protein drugs as well as possible anti-cancer mechanisms. Further research is on going to determine the bioactive principle(s) of recombinant protein RPL23A responsible for its anticancer activity.
The present study investigated the structural characterization and immune regulation of a novel polysaccharide from Maerkang Lactarius deliciosus Gray. Chemical methods, high performance gel permeation chromatography, fourier transform infrared spectroscopy, nuclear magnetic resonance spectrum and gas chromatography-mass spectrometry were used to characterize the polysaccharide structure. The immunomodulatory abilities of the Maerkang L deliciosus Gray polysaccharide (LDG-M) were also investigated. LDG-M was primarily composed of beta-D-glucose and alpha-D-lyxose with the ratio of 2:1. The possible structure of LDG-M had a backbone of 1,6-linked-beta-D-glucose and 1,4,6-linked-beta-D-glucose, with branches primarily composed of one (1 -> 4)-linked-alpha-D-lyxose residue. The immunoregulatory activity results demonstrated that LDG-M promoted the proliferation and phagocytosis of macrophages, and induced cytokine release. LDG-M also promoted the proliferation of B cells by affecting the G(0)/G(1), S and G2/M phases. The present study introduced LDG-M as a valuable source with unique immunoregulatory properties.
1 February 2018 in International Journal of Molecular Medicine
A new heteropolysaccharide was extracted and purified from the fruiting bodies of Cantharellus cibarius Fr. The Cantharellus cibarius Fr. polysaccharide (CC-1) had a molecular weight of 61,056 kDa and was mainly formed of the glucose and xylose at ratio of 5:1. Structure identification of CC-1 was analysed by a combined application of total hydrolysis, high performance liquid chromatography (HPLC), methylation analysis, gas chromatography-mass spectrometry (GC-MS), infrared (IR) spectra and nuclear magnetic resonance (NMR) spectroscopy. The experimental results showed that CC-1 had a backbone of 1,4-linked-β-D-glucose which branched at O-6 and the branches were mainly composed of 6→1)-α-D-xylopyranose residue. CC-1 exhibited significant in vitro antioxidant effect and proliferation effect of immune cells. The activity study showed CC-1 has ability to clear the ABTS+ free radical and DPPH- free radical in a certain range of concentration. The proliferation activity of the immune cells showed that the proliferation effect on B cells was very significant (P<0.001) in the concentration of 0.625-80 mg/ml; and the effect of T cell proliferation was also very significant (P<0.001) in the concentration of 5-20 mg/ml. The result of this study introduced Cantharellus cibarius Fr. as a possible valuable source in exhibiting unique immunoregulatory and antioxidant properties.
About Me
Yiling Hou, mainly engaged in biochemistry and molecular biology research.
Yiling Hou, mainly engaged in biochemistry and molecular biology research.