Chemical modification, characterization and bioactivity of Chinese lacquer polysaccharides from lac tree Rhus vernicifera against leukopenia induced by cyclophosphamide

Lacquer polysaccharide (LP) was isolated from the sap of lac tree (Rhus vernicifera). Its derivatives, carboxymethyl LP, sulfated LP and debranching LP were prepared. Their structure was analyzed by GPC, FT-IR and NMR spectroscopy. The sugar components of carboxymethyl and sulfated LPs hardly changed, but the molecular weight of the former decreased. The side chains of LPs were partially removed using sodium periodate in mild conditions and the pyranose ring β-configuration of products obtained was not changed. Bioactivity of natural and modified LPs against leukopenia induced by cyclophosphamide (CP) was investigated in mice. LP exhibited a significant bioactivity (P<0.05) compared to positive control group (CP). The bioactivity could increase slightly with the increasing of the contents of carboxymethyl groups. However, with the removal of the side chains and the incorporation of sulfate groups, the bioactivity gradually decreased. These showed that the bioactivity of lacquer polysaccharides against leukopenia induced by CP was strongly dependent on the types of ionic groups of the polysaccharides and concerned with the side chains with 4-O-methyl-β-glucuronic acid in the terminal.     

The structure-anticoagulant activity relationships of sulfated lacquer polysaccharide: effect of carboxyl group and position of sulfation

Regiospecific oxidation of the primary hydroxyl groups in lacquer polysaccharide (LPL, Mw 6.85 x 10(4)) and its NaIO4 oxidation derivatives (LPLde) to C-6 carboxy groups was achieved with NaOCl in the presence of Tempo and NaBr. Sulfate groups were incorporated into the oxidated polysaccharides using Py.SO3 complex as a reagent. Reactivity of polysaccharide hydroxyl group was C-6 > C-2 > C-4. Sulfate groups were mainly linked to the second hydroxy at C-2 in the products. The results of APTT assay showed after incorporation of carboxyl groups into lacquer polysaccharides, the intrinsic coagulation pathway was promoted, and all sulfated polysaccharides had very weak anticoagulant activity within the scope of studied DS (0.39-1.11). These indicated that carboxyl groups and sulfate groups had the synergistic action. At the same time, the anticoagulant activity increased very slowly with the DS in the second hydroxy. This indicated that 6-O-SO3- in the side chains took an important role in the anticoagulant activity.     

Sulfation of Chinese lacquer polysaccharides in different solvents

A branched ionic polysaccharide isolated from the sap of the Chinese lac tree (Rhus vernicifera) was chemically modified by sulfation using sulfur trioxide–pyridine (SO₃·Py) complex as a reagent. Effects of molar ratio of SO₃·Py complex to sugar unit, reaction time and reaction temperature on degree of sulfation (DS) and molecular weights of products were studied. Solvent was another important factor affecting sulfation reaction. In different solvents, when the other conditions remained constant, DS and molecular weights were in the following order: DMSO>DMF>FA (formamide) and DMSO3·Py complex. Based on these, we deduced that degradation of polysaccharide in the sulfation reaction process involved both dehydrolysis and hydrolytic degradation.     

Chemical modification of alpha-thrombin and in vitro characterization of its anticoagulant activity

Acetylthrombin, succinylthrombin, maleylthrombin, polyethylene glycol (PEG)-thrombin, and acetylated PEG-thrombin were prepared and characterized. Although clotting activities could be reduced remarkably by the modification, the potencies of all the modified thrombins for protein C activation also decreased. However, in the case of acetylthrombin, succinylthrombin, and acetylated PEG-thrombin, the remaining potencies for protein C activation exceeded largely the remaining clotting activities. Moreover, by the addition of thrombodulin, the potencies for protein C activation were enhanced 16 - 60 fold for succinylthrombin, maleylthrombin, and PEG-thrombin.     

Structure and molecular weight of Asian lacquer polysaccharides

Structural analysis of Asian lacquer polysaccharides in Vietnam, Myanmar, Cambodia, Taiwan, and Japan was carried out by a combination of chemical and physical methods, and then their structures were compared with that of a Chinese lacquer polysaccharide reported previously. It was found that the structure of polysaccharides in China and Japan, Taiwan and Vietnam, Myanmar and Cambodia, was similar to each other. The polysaccharides in Myanmar and Cambodia had larger amounts of -arabinose and -rhamnose than those in other Asian lacquer polysaccharides. In addition, the degradation process of lacquer polysaccharide was revealed for the first time by the time-course of GPC measurements of polysaccharide in Aizu, Japan. The results suggest that the molecular weight of polysaccharide in lacquer tree had around with narrow molecular weight distribution and then decreased gradually into two molecular weight fractions of and 23×10³ in the proportion of 25 and 75 mol% isolated after 3 weeks.     

肠浒苔多糖的羧甲基化修饰及其抗氧化活性研究

该研究采用氢氧化钠-氯乙酸的化学反应体系制备羧甲基化肠浒苔多糖,以获得不同取代度的羧甲基化肠浒苔多糖,取代度的大小受氢氧化钠浓度、反应温度和反应时间的影响。结果表明:(1)当氢氧化钠浓度20%、反应温度60℃、反应时间3 h时,得到羧甲基化的最大取代度为0.781。(2)通过体外抗氧化来评价不同羧甲基化肠浒苔多糖的抗氧化活性。(3)当羧甲基化肠浒苔多糖的浓度为1.6 mg·mL~(-1)时,羧甲基化肠浒苔多糖清除羟基自由基、超氧阴离子自由基的能力分别为44.45%、51.98%,其清除DPPH自由基清除率和还原能力分别为16.75%、0.457 6。(4)与修饰前的相比,羟基自由基、超氧阴离子的清除能力均有较大幅度提高,羧甲基化修饰对肠浒苔多糖的DPPH自由基和还原力有减弱作用。以上结果表明,羧甲基化修饰引起的肠浒苔多糖的结构变化可以提高其抗氧化活性。     

Chemical modification of nodulisporic acid A: preliminary structure-activity relationships

Medicinal chemistry efforts were initiated to identify the key constituents of the nodulisporic acid A (1) pharmacophore that are integral to its potent insecticidal activity. New semisynthetic derivatives delineated 1 into 'permissive' and 'nonpermissive' regions and led to the discovery of new nodulisporamides with significantly improved flea efficacy.     

Chemical modification of chloroperoxidase for enhanced stability and activity

Chloroperoxidase from Caldariomyces fumago (CPO, EC 1.11.1.10) is one of the most interesting enzymes from the group of heme peroxidases and has been extensively applied in synthetic processes. Nevertheless, the practical application of CPO is limited due to its very low operational stability, especially in the presence of peroxidative compounds. For this reason, effect of chemical modifications of CPO in the stability of the enzyme was studied. Side-chain selective modifications of amino groups of Lys residues, and carboxyl groups of Asp and Glu residues, as well as crosslinking and periodate oxidation of sugar moiety were carried out. The stability of modified CPOs was evaluated at elevated pH and temperature, and in the presence of tert-butyl hydroperoxide. Effect of modification of CPO on the performance of the reaction of Cbz-ethanolamine oxidation was studied as well. Those modifications that involved carboxyl groups via carbodiimide coupled method and the periodate oxidation of the sugar moiety produced better catalysts than native CPO in terms of stability and activity at elevated pH values and temperatures.     

Sulfated modification can enhance the immune-enhancing activity of lycium barbarum polysaccharides

In test in vitro, four sulfated lycium barbarum polysaccharides (sLBPSs) with different degrees of sulfation (DS), sLBPS_0.7, sLBPS_1.1, sLBPS_1.5 and sLBPS_1.9, were added into cultured chicken peripheral lymphocytes and the changes of lymphocytes proliferation were compared by MTT assay taking the non-modified LBPS as control. Two sLBPSs with better efficacy, sLBPS_1.5 and sLBPS_1.9 were selected. In test in vivo, one hundred 14-day-old chickens were averagely divided into five groups randomly. The chickens except blank control group were vaccinated with Newcastle disease vaccine, repeated vaccination at 28 days old. At the same time of the first vaccination, the chickens in three experimental groups were injected with 0.5 mL of sLBPS_1.5, sLBPS_1.9 and LBPS at 4 mg mL⁻¹, in vaccination control group, with 0.5 mL of physiological saline, once a day for three successive days. On days 7, 14, 21 and 28 after the first vaccination, the changes of peripheral lymphocytes proliferation and serum HI antibody titer were determined. The result showed that two sLBPSs could significantly promote lymphocytes proliferation and enhance serum antibody titer. These results indicated that sulfated modification could enhance the immune-enhancing activity of LBPS, which there was a certain relativity with the DS of sulfated polysaccharide. sLBPS_1.5 possessed the best efficacy and would be expected as the component drug of a new-type immunopotentiator.     

Chemical modification of the carbohydrate moiety in N-acetylmuramoyl-l-alanyl-d-isoglutamine,and the immunoadjuvant activity

Five carbohydrate analogs of N-acetylmuramoyl-l-alanyl-d-isoglutamine have been synthesized from benzyl 2-acetamido-2-deoxy-3-O-[d-1-(methoxycarbonyl)ethyl]-α-d-glucopyranoside (1) and the corresponding 6-O-benzoyl derivative (2). Chlorination of 1 and 2 with triphenylphosphine in carbon tetrachloride gave the 4,6-dichloro compound 3 and the 6-O-benzoyl-4-chloro compound (4), which were treated with tributyltin hydride, to yield benzyl 2-acetamido-2,4,6-trideoxy-3-O-[d-1-(methoxycarbonyl)ethyl]-α-d-xylo-hexopyranoside (6) and benzyl 2-acetamido-6-O-benzoyl-2,4-dideoxy-3-O-[d-1-(methoxycarbonyl)ethyl]-α-d-xylo-hexopyranoside (7), respectively. Methanesulfonylation of 8, derived from 7 by debenzoylation, gave the 6-methanesulfonate, which underwent displacement with azide ion to afford benzyl 2-acetamido-6-azido-2,4,6-trideoxy-3-O-[d-1-(methoxycarbonyl)ethyl]-α-d-xylo-hexopyranoside (10). Hydrolysis of the methyl ester group in compounds 3, 5 (debenzoylated 4), 6, 8, and 10 gave the corresponding free acids, which were coupled with l-alanyl-d-isoglutamine benzyl ester, to yield the dipeptide derivatives in excellent yields. Hydrogenation of the dipeptide derivatives thus obtained gave the five carbohydrate analogs of N-acetylmuramoyl-l-alanyl-d-isoglutamine, respectively, in good yields. The immunoadjuvant activity of the N-acetylmuramoyl-dipeptide analogs was examined.     

Chemical characterization, antiproliferative and antiadhesive properties of polysaccharides extracted from Pleurotus pulmonarius mycelium and fruiting bodies

Mushroom polysaccharides are potent substances that exhibit antitumor and immunomodulatory properties. Studies comparing the chemical composition and antitumor-related activities of polysaccharides released by fungal strains under different growth conditions are not available. Thus, the present study compared polysaccharides extracts produced by Pleurotus pulmonarius from mycelium grown in liquid culture (ME) or fruiting bodies (FBE). Polysaccharides of both ME and FBE had a relatively high molecular mass. NMR spectroscopy indicated that ME glucan is an α-glucan whereas FBE glucan is a mixture of both α- and β-glucans. Glucose and galactose where the most prominent monosaccharide in both glucans. Treatment of several colon cancer cell lines expressing varying amounts of galectin-3 with the two fungal glucans inhibited their viability and significantly reduced their ability to adhere to the key component of the extracellular matrix, fibronectin, and to a human umbilical vein endothelial cell monolayer, in a time- and dose-dependent manner mainly in those cell lines expressing high amounts of galectin-3. We conclude that ME and FBE glucans may exert a direct antiproliferative effect on cancer cells expressing high galectin-3 concentrations and concomitantly downregulate tumor cell adherence, the latter being directly related to cancer progression and metastasis.     

Enzymatic modification of plant polysaccharides

Enzymes find widespread industrial use in the modification of the functional properties of plant polysaccharides both in vivo and in vitro. Reactions catalysed include depolymerization, debranching and de-esterification, depending on the specific enzyme or enzyme mixture employed and on the particular industrial requirement. Depolymerization of pentosans and/or barley β-glucans to destroy their viscosity-building properties is essential in starch and gluten manufacture, in the mashign of barley malt and in the production of maltosaccharide syrups. Depolymerization of pectin is required in juice clarification and to allow concentration. However, in other instances the aim may be to maintain or, at most, only slightly alter the molecular size of functional polysaccharides, i.e. in the conversion of guar galactomannan to a locust-bean type galactomannan and in the enzymic treatment of wheat-flour doughs. Enzymes may also be used to produce specific oligosaccharide fragments from polysaccharides and as diagnostic tools in the measurement of a particular polysaccharide in a mixture.     

Purification and characterization of hydrosoluble components from the sap of Chinese lacquer tree Rhus vernicifera

Continuous gradient elution chromatography (CGEC) was employed to purify and separate enzymes and polysaccharides from the sap of Rhus vernicifera Chinese lacquer tree. There are three different molecules with laccase enzyme activity. Two are enzymes of each other (L1, and L2), whereas the third (RL) is an entirely separate entity. Two polysaccharides (GP1 and GP2) were also found. The Rhus laccase (RL), and isoenzymes L1 and L2, have peak molecular masses of 109,100, 120,000, 103,000 respectively; each has four copper atoms per molecule, and the pI values were 8.2, 8.6, and 9.1, respectively. The structure of the laccases was studied by Fourier-transform infrared (FT-IR) and Matrix-assisted laser desorption/ionization time-of flight (MALDI-TOF) mass spectrometry. The typical amide I (1646 cm⁻¹) and amide II (1545 cm⁻¹) bands were observed. The results from MALDI-TOF were similar to those from CGEC, but the molecular mass from the MALDI-TOF was significantly different from that obtained from sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).     

Chemical modification of the glucosidase inhibitor 1-deoxynojirimycin. Structure-activity relationships.

The ability of the glucosidase inhibitor 1-deoxynojirimycin (dNM) and a series of N-alkylated dNM derivatives to interfere with biosynthesis, transport, and maturation of the glycoprotein alpha 1-antitrypsin in HepG2 cells was investigated. Inhibition of endoplasmic reticulum glucosidase I and II by dNM and its derivatives resulted in an intracellular accumulation of alpha 1-antitrypsin with glucose-containing high mannose type oligosaccharides (precursor). N-alkylation of dNM increased its potency in inhibiting endoplasmic reticulum glucosidases, as determined from the concentration required for half maximal inhibition. N-Alkylated derivatives of dNM were better able to inhibit glucosidase I than glucosidase II (deduced from the number of glucose residues retained in Endo H-releasable oligosaccharides). The inhibition of glucosidase activity imposed by alkylated dNM derivatives was less easily reversed than that by dNM, an effect most pronounced for N-methyl-dNM. Branching of the alkyl group of dNM derivatives decreased the inhibitory potency. Although dNM and its derivatives interfered strongly with intracellular oligosaccharide processing, they did not completely block N-glycan maturation of alpha 1-antitrypsin even at the highest concentrations tested.     

Extraction optimization,physicochemical characterization,and antioxidant activity of polysaccharides from Rhodosorus sp. SCSIO-45730

Journal of Applied Phycology - Microalgal polysaccharides have been reported in many studies due to their uniqueness, biocompatibility, and high value, and Rhodosorus sp. SCSIO-45730 was an...     

Chemical synthesis of polysaccharides

Here we summarize the chemical syntheses of glycans containing over 20 monosaccharide units, with the recent syntheses of ultra-long glycans, including a 92mer arabinogalactan, a 100mer and a 151mer mannan, and a 128mer rhamnomannan, being highlighted. The assembly strategies, glycosylation methods, and protecting group manipulations, which are crucial to the successful synthesis, are discussed to give guidance for the future synthesis of various polysaccharides.     

Chemical characterization and genetic relationships among Ocimum basilicum L. cultivars

Twenty-seven Ocimum basilicum cultivars were subjected to a chemical characterization of essential oil components by gas chromatography/mass spectrometry (GC/MS) and a genetic characterization using the amplified fragment-length polymorphism (AFLP) technique. Since the same 27 accessions had previously been classified into six morphotypes, these analyses allowed us to make detailed comparisons of chemistry, genetics, and morphology. The chemical composition and morphology of the studied cultivars appeared to have a strong genetic component. The AFLP analysis revealed a distinction between the green and purple morphotypes. The green morphotypes predominantly utilized the terpene biosynthetic pathway, while most purple morphotypes primarily utilized the phenylpropene biosynthetic pathway. The GC/MS analysis led to identification of 87 volatiles. Among the 27 cultivars, five chemotypes were identified. A detailed characterization of the essential oil constituents indicated the existence of both specific combinations of compounds and 'private' compounds with the potential to be used in many aspects of human life. The established relationship between a genetic profile, chemical composition, and morphology represents an important step in future breeding programs and in the cultivation of this species.