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1.
Abstract: The water‐soluble crude polysaccharide tea flower polysaccharide (TFP), obtained from tea (Camellia sinensis) flower by boiling‐water extraction and ethanol precipitation, was fractionated by Sephadex G‐100 column chromatography, giving 2 polysaccharide fractions termed TFP‐1 and TFP‐2. The structural features of TFP‐1 and TFP‐2 were investigated by high‐performance liquid chromatography (HPLC), gel‐permeation chromatography (GPC), rheometer, infrared (IR) spectra, nuclear magnetic resonance (NMR) spectroscopy, atomic force microscope (AFM), and scanning electron microscope (SEM). Results indicated that TFP‐1 was composed of glucose: xylose: rhamnose: galactose = 1.0:1.2:0.81:0.98 with a molecular weight of 167.5 KDa, while TFP‐2 comprised glucose: xylose: rhamnose: arabinose = 1.0:0.76:2.3:2.3 with a molecular weight of 10.1 KDa. The 1H NMR revealed that TFP‐1 contained α‐L‐Rhap, α‐D‐Galp, α‐D‐GalpNAc, α‐D‐Xylp, α‐D‐Glcp, and β‐D‐Glcp residues, while TFP‐2 was illustrated to have α‐L‐Rhap, α‐L‐Arap, α‐D‐Xylp, α‐D‐Glcp, and α‐D‐GlcpNAc residues. Antioxidant activities of these fractions were investigated using various in vitro assay systems compared with ascorbic acid. In conclusion, TFP‐2 exhibited the higher antioxidant activities and could be explored as a novel potential antioxidant. Practical Application: At present, commonly low‐grade tea is preferred to extract the tea polysaccharide, to take full advantage of tea flower resource to extract polysaccharides can greatly reduce the cost of tea products. Thus, the search for plant‐derived biomaterials from this study could generate natural value‐added products from underutilized tea plant waste and used as a medicinal agent against chronic health problems, such as cancers, aging, and atherosclerosis caused by reactive free radicals that produced from oxidation.  相似文献   

2.
BACKGROUND: To contribute towards understanding the relationship of structure and bioactivity, a protein‐bound acidic polysaccharide named TPC3‐1 was isolated and purified from low‐grade green tea (Camellia sinensis L.). The homogeneity and weight average molecular weight of TPC3‐1 was determined by agarose gel electrophoresis and high‐performance gel permeation chromatography. The monosaccharide and amino acid composition of TPC3‐1 were analysed by gas chromatography and an amino acid analyser. The molecular structure of TPC3‐1 was characterised by Fourier transform infrared spectroscopy, 13C nuclear magnetic resonance spectroscopy and atomic force microscopy. RESULTS: Based on the data obtained, the average peak molecular weight of TPC3‐1 was about 120 kDa. TPC3‐1 was composed of L ‐arabinose, D ‐ribose, D ‐xylose, D ‐glucose and D ‐galactose with a molar ratio of 4.9:2.2:3.1:1.8:1.0. Fifteen amino acids were identified as components of the polymer. The TPC3‐1 molecule was found to have an anomeric carbon sign of both α and β configurations and high‐branched chains. The network structure of TPC3‐1 was observed. CONCLUSION: The tea polysaccharide TPC3‐1 was an acid protein‐bound polysaccharide with an image of network structure. The results presented here will facilitate further study of the relationship between the chemical structure and biological role of tea polysaccharide. Copyright © 2008 Society of Chemical Industry  相似文献   

3.
Phellinus linteus is a medicinal mushroom that has been used in Oriental countries for centuries for its antitumor, antioxidant, immunomodulatory, and biological activity on hyperglycemia. A water‐soluble crude polysaccharide was extracted using hot water from P. linteus mycelia grown under submerged culture. An orthogonal experiment was used to optimize the extraction conditions of P. linteus mycelia polysaccharides (PLP). The crude polysaccharide was purified using DEAE Sephadex A‐50 and Sephadex G‐200 chromatography. Fourier transform infrared (FT‐IR) spectroscopy and nuclear magnetic resonance (1H NMR) spectroscopy were used to investigate the structure of the purified P. linteus polysaccharide (PLP‐I), revealing that it was mainly a branched‐type glycan with both α‐ and β‐linkages and a pyranoid sugar ring conformation. PLP orally administered at 100 mg/kg body weight/d could significantly reduce the blood glucose level by 35.60% in alloxan‐induced diabetic mice. The results of an oral glucose tolerance test (OGTT) revealed that PLP had an effect on glucose disposal after 28 d of treatment. The result revealed that PLP from a submerged culture of P. linteus mycelia possessed potent hypoglycemic properties. The polysaccharide may be useful as a functional food additive and a hypoglycemic agent.  相似文献   

4.
BACKGROUND: Recently, much attention has focused on the biological properties of fungal polysaccharides. Polysaccharides extracted from Phellinus baumii Pilát are reported to have antitumor and immunostimulatory properties, as well as anti‐mutagenicity activity, hypoglycemic and free radical scavenging properties. In this paper, we report the chemical and structural characterization of a novel neutral polysaccharide isolated from the fruiting bodies of P. baumii Pilát. RESULTS: PBF4, a purified polysaccharide, was isolated from the fruiting bodies of P. baumii Pilát, and was shown to be composed of L ‐fucose and D ‐glucose in a ratio of 1:4. It was found to be a fucoglucan consisting of a α‐(1 → 4)‐D ‐glucopyranose backbone with some insertions of α‐(1 → 2)‐L ‐Fucp residues. It also contained a minor β‐(1 → 4,6)‐linked D ‐glucopyranosyl and β‐glycosidically linked nonreducing‐end D ‐glucopyranosyl residues. CONCLUSION: Our results add a new polysaccharide to those already described for different fungal groups. This kind of polysaccharide is useful for further study of the structure–function relationships and mechanism responsible for its biological activities. Copyright © 2008 Society of Chemical Industry  相似文献   

5.
BACKGROUND: Polysaccharides extracted from fungi or algae have shown a variety of medical activities, and the exploitation of polysaccharides for application in pharmacy has been very promising. In this study, the immunomodulatory properties of polysaccharides from Morchella esculenta were investigated to identify immunostimulatory activity and potentially contribute to the therapeutic potential of Morchella esculenta. RESULTS: A water‐soluble polysaccharide, MEP, was obtained from the fermentation broth of Morchella esculenta. Two fractions of this polysaccharide (MEP‐I and MEP‐II) were extracted and purified. High‐performance gel permeation chromatography analysis showed the average molecular weights of two polysaccharides. Structural properties and compositions of these two fractions were examined by Fourier transform infrared and a high‐performance liquid chromatography with an evaporative light scattering detector. Active experiments suggested that the MEP had typical immunostimulatory activity. CONCLUSION: These bioactivity tests showed that the polysaccharides from Morchella esculenta presented significant immune modulating activity, and this finding may offer the basis for the popular use of polysaccharides in functional foods or medicine. Copyright © 2011 Society of Chemical Industry  相似文献   

6.
Four types of carbohydrates, including Dendrobium officinale polysaccharide, Dendrobium aphyllum polysaccharide and β‐glucans from yeast and barley, were examined, and their structures were found to mainly contain 1,4‐linked‐β‐d ‐Glcp. Artificially simulated gastrointestinal digestion was conducted to characterise the changes of molecular weight, reducing sugars and released free monosaccharides by high‐performance liquid chromatography, kits and the newly developed gas chromatography (GC)‐mass spectrometry (MS)/MS analysis, which indicated that high molecular weight and complex spatial structures contributed to delayed monosaccharide release following exposure to digestive solution. The spatial structures of carbohydrates were changed during gastric digestion, but their primary structures were destroyed during intestinal digestion. Additionally, for the developed 7890A/7000 GC‐TQ/MS‐MS, the new analytical method was successfully used to analyse very low concentrations of monosaccharides in the simulated gastrointestinal digestive system.  相似文献   

7.
An aminopeptidase (AP) was partially purified from jumbo squid (Dosidicus gigas) hepatopancreas with 154.24‐fold and yield of 6.15%. The purification procedure consisted of ammonium sulphate fractionation and DEAE‐Sephacel chromatography. The enzyme was approximately 48–53 kDa as estimated by SDS‐PAGE. With l ‐leu‐p‐NA, it had optimum activity at pH 8.0 and 30 °C. The Km and Vmax/Km values of the enzymes for l ‐leu‐p‐NA were 0.326 mm and 2787 at 37 °C, respectively. Activation energy (Ea) of the enzyme was 53.50 kJ M?1.The AP showed activity against seven synthetic substrates: l ‐proline>l ‐methionine>Ac. l ‐γ‐glutamic>l ‐glycine>l ‐leucine>l ‐alanine>l ‐lysine‐p‐NA. The enzyme was strongly inhibited by Bestatin, partially inhibited by a metal‐chelating agent and by PCMB, a cystein protease inhibitor. Zn2+ and (or) Ca2+ seemed to be its metal cofactor(s). Incubation of casein with the partially purified AP resulted in a degree of hydrolysis of 6%.  相似文献   

8.
BACKGROUND: Seed galactomannans are preferred hydrocolloids since they are comparatively cheap, non‐toxic, eco‐friendly and non‐polluting during production and application. Galactomannans from seeds of three species of Gleditsia, namely G. sinensis, G. microphylla and G. melanacantha, were characterized in terms of structural and thermal properties. RESULTS: Gleditsia polysaccharides were characterized using both chemical and chromatographic methods, as well as Fourier transform infrared, 1H nuclear magnetic resonance (NMR) and 13C NMR spectroscopy, and it was shown that they consist of D ‐mannopyranose and D ‐galactopyranose residues. The mannose/galactose (M/G) ratio of galactomannans was 3.25, 3.31 and 2.30, respectively. It was also found that these polysaccharides differ from one another in values of Mw, Mn and polydispersity. X‐ray diffraction confirmed the amorphous nature of Gleditsia galactomannans, although G. sinensis galactomannan showed a high crystallinity. Thermal analysis of the galactomannans by differential scanning calorimetry illustrated that their endothermic peaks ranged from 290 to 320 °C. CONCLUSION: Gleditsia polysaccharides are neutral galactomannans. The higher value of M/G ratio from G. sinensis and G. microphylla indicates that their gums offer an excellent alternative for locus bean gum. Copyright © 2011 Society of Chemical Industry  相似文献   

9.
An α‐l ‐rhamnosidase secreted by Penicillium citrinum MTCC‐8897 has been purified to homogeneity from the culture filtrate of the fungal strain using ammonium sulphate precipitation and cation‐exchange chromatography on carboxymethyl cellulose. The sodium dodecyl sulphate/polyacrylamide gel electrophoresis analysis of the purified enzyme gave a single protein band corresponding to the molecular mass 51.0 kDa. The native polyacrylamide gel electrophoresis also gave a single protein band confirming the enzyme purity. The Km and Vmax values of the enzyme for p‐nitrophenyl α‐l ‐rhamnopyranoside were 0.36 mm and 22.54 μmole min?1 mg?1, respectively, and kcat value was 17.1 s?1 giving kcat/Km value of 4.75 × 104 m ?1 s?1. The pH and temperature optima of the enzyme were 7.0 and 60 °C, respectively. The purified enzyme liberated l ‐rhamnose from naringin, rutin, hesperidin and wine, indicating that it has biotechnological application potential for the preparation of l ‐rhamnose and other pharmaceutically important compounds from natural glycosides containing terminal α‐l ‐rhamnose and also in the enhancement of wine aroma.  相似文献   

10.
BACKGROUND: In Tunisia, sardinelle (Sardinella aurita) catches totalled about 13 300 t in 2002. During processing, solid wastes including heads and viscera are generated, representing about 30% of the original raw material. Viscera, one of the most important by‐products of the fishing industry, are recognised as a potential source of digestive enzymes, especially proteases with high activity over a wide range of pH and temperature conditions. This paper describes the purification procedure and some biochemical characterisation of trypsin from S. aurita viscera. RESULTS: Trypsin from the viscera of sardinelle (S. aurita) was purified by fractionation with ammonium sulphate, Sephadex G‐75 gel filtration, Sepharose mono Q anion exchange chromatography, ultrafiltration and a second Sephadex G‐75 gel filtration, resulting in a 5.42‐fold increase in specific activity and 6.1% recovery. The molecular weight of the purified enzyme was estimated to be 24 kDa using size exclusion chromatography and sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified enzyme showed esterase‐specific activity on N‐α‐benzoyl‐L ‐arginine ethyl ester (BAEE) that was four times greater than its amidase‐specific activity on N‐α‐benzoyl‐DL ‐arginine‐p‐nitroanilide (BAPNA). The optimal pH and temperature for enzyme activity were pH 8 and 55 °C respectively using BAEE as a substrate. The trypsin kinetic constants Km and kcat on BAPNA were 1.67 mmol L?1 and 3.87 s?1 respectively, while the catalytic efficiency kcat/Km was 2.31 s?1 L mmol?1. CONCLUSION: Trypsin was purified from sardinelle (S. aurita) viscera. Biochemical characterisation of S. aurita trypsin showed that this enzyme can be used as a possible biotechnological tool in the fish‐processing and food industries. Copyright © 2008 Society of Chemical Industry  相似文献   

11.
An extracellular α‐l ‐rhamnosidase from Penicillium citrinum MTCC‐3565 has purified to homogeneity from its culture filtrate using ethanol precipitation and cation‐exchange chromatography on carboxymethyl cellulose. The purified enzyme gave a single protein band corresponding to molecular mass of 45.0 kDa in SDS‐PAGE analysis showing the purity of the enzyme preparation. The native PAGE analysis showed the monomeric nature of the purified enzyme. Using p‐nitrophenyl α‐l ‐rhamnopyranoside as substrate, Km and Vmax values of the enzyme were 0.30 mm and 27.0 μm min mg?1, respectively. The kcat value was 20.1 s giving kcat/Km value of 67.0 mm s?1 for the same substrate. The pH and temperature optima of the enzyme were 8.5 and 50 °C, respectively. The activation energy for the thermal denaturation of the enzyme was 29.9 KJ mol?1. The α‐l ‐rhamnosidase was able to hydrolyse naringin, rutin and hesperidin and liberated l ‐rhamnose, indicating that the purified enzyme can be used for the preparation of α‐l ‐rhamnose and pharmaceutically important compounds by derhamnosylation of natural glycosides containing terminal α‐l ‐rhamnose. The α‐l ‐rhamnosidase was active at the level of ethanol concentration present in wine, indicating that it can be used for improving wine aroma.  相似文献   

12.
Potato proteins (PP) in the lab‐made potato juice consists of patatins (38.1%), protease inhibitors (56.7%) and other high molecular weight proteins (5.1%) as characterised by Tricine‐SDS‐PAGE and MALDI‐TOF/TOF‐MS. To recover PP from potato juice, the complex behaviours of PP with several natural polyelectrolytes as a function of pH and protein to polyelectrolyte ratio (RPP/Polysaccharide) were studied by turbidimetric titration. Results indicated that the turbidity curves of PP‐polysaccharides displayed a shift to higher pH for chitosan and lower pH for anionic polysaccharides with the decreasing of RPP/Polysaccharide. Chitosan could be used to selectively recover patatin with the purity of 88.6%, and the highest protein yield (51.9%) was achieved at pH 6.0 and RPP/chitosan of 1:2. Carrageenan, one typical anionic polysaccharide, could recover all the PP in potato juice at pH 3.5 and RPP/carrageenan of 2.5:1 with no special protein selectivity.  相似文献   

13.
The extracellular α‐l ‐rhamnosidase has been purified by growing a new fungal strain Aspergillus awamori MTCC‐2879 in the liquid culture growth medium containing orange peel. The purification procedure involved ultrafiltration using PM‐10 membrane and anion‐exchange chromatography on diethyl amino ethyl cellulose. The purified enzyme gave single protein band in SDS‐PAGE analysis corresponding to molecular mass 75.0 kDa. The native PAGE analysis of the purified enzyme also gave a single protein band, confirming the purity of the enzyme. The Km and Vmax values of the enzyme for p‐nitrophenyl‐α‐l ‐rhamnopyranoside were 0.62 mm and 27.06 μmole min?1 mg?1, respectively, yielding kcat and kcat/km values 39.90 s?1 and 54.70 mm ?1 s?1, respectively. The enzyme had an optimum pH of 7.0 and optimum temperature of 60 °C. The activation energy for the thermal denaturation of the enzyme was 35.65 kJ?1 mol?1 K?1. The purified enzyme can be used for specifically cleaving terminal α‐l ‐rhamnose from the natural glycosides, thereby contributing to the preparation of pharmaceutically important compounds like prunin and l ‐rhamnose.  相似文献   

14.
Water‐soluble polysaccharides from coarse green tea were separated by anion‐exchange chromatography into five fractions (fraction A [FA], fraction B, fraction C [FC], fraction D and fraction E). Two of these fractions, FA and FC, contained significant glucokinase‐stimulating activity (P < 0.05). The major component, FC, showed the most activity, and thus, was further purified by gel filtration chromatography, thereby obtaining fraction C‐1 (FC‐1) and fraction C‐2 (FC‐2). The biological activity of the two fractions was investigated, and FC‐1 displayed higher glucokinase‐stimulating activity (P < 0.01). Chemical tests combined with IR and UV spectroscopy revealed that FC‐1 is an acidic polysaccharide without conjugation to protein. Sugars of FC‐1 are composed of rhamnose, arabinose, mannose, glucose and galactose in the ratio of 12.57:22.95:4.4:39.34:20.77. Uronic acid analysis by ion chromatography showed that FC‐1 contains 8% galacturonic acid, and its molecular weight was estimated to be approximately 6 × 104 using a Sephacryl S200 column. These results are different from the observations previously reported, therefore suggesting that FC‐1 is a novel polysaccharide.  相似文献   

15.
Fungal polysaccharides are novel ingredients in functional foods with diverse medicinal properties. Here, a polysaccharide, MSP2-1, was isolated from Morchella sextelata fruiting bodies and purified using column chromatography. MSP2-1 (371.5 kDa) is a branched (1 → 4)-α-D-glucan with the O-6 or O-3 position occupied by α-D-Glcp. Light scattering analysis revealed that MSP2-1 has a spherical conformation in 0.9% NaCl solution; this was confirmed using transmission electron microscopy. Finally, MPS2-1 was found to promote proliferation, NO release and cytokine secretion in RAW264.7 cells through a mechanism involving Toll-like receptor 4. Collectively, these results highlight the potential application of MSP2-1 as an immunoenhancing compound for hypoimmunity.  相似文献   

16.
In this work, crude polysaccharide extracts were extracted from pumpkin (Cucurbita moschata) fruit by hot water. After removal of proteins and purification, polysaccharides of pumpkin fruit (PP1‐1) were subjected to structural identification. Gas chromatography analysis indicated that PP1‐1 comprised of galactose (86.4%), and glucose (13.6%). The molecular weight of PP1‐1 was measured to be 0.87 × 104 Da by gel permeation chromatography. The inhibitory kinetic evaluation showed that it was non‐competitive inhibition of PP1‐1 on the α‐glucosidase‐catalysed hydrolysis of PNPG. The Michaelis–Menten constant (Km) was 0.106 m , and the inhibitory constants (Ki), 0.435 mg.  相似文献   

17.
The objective of this study was to investigate the multiple relations between the preliminary molecular structural characteristics and antioxidant activities of polysaccharides from Canarium album (Lour.) Raeusch (CPS). Three polysaccharide fractions, CPS1, CPS2, and CPS3, were isolated from CPS by column chromatography. CPS1 and CPS3 were mainly composed of neutral polysaccharides linked by α‐ and β‐glycosidic linkages while CPS2 was pectin polysaccharides mainly linked by β‐glycosidic linkages. According to the SEC‐MALLS‐RI system, the molecular weight of CPS1 was greater compared to CPS2 and CPS3, and the molecular weight and radius of CPS did not display positive correlation. The chain conformation analysis indicated CPS1 and CPS2 were typical highly branched polysaccharides while CPS3 existed as a globular shape in aqueous. Furthermore, the antioxidant activity of CPS2 was better than that of CPS3, while that of CPS1 was the weakest. The antioxidant activities of polysaccharide fractions were affected by their monosaccharide composition, glycosidic linkage, molecular weight, and chain conformation. This functional property was a result of a combination of multiple molecular structural factors. CPS2 was the major antioxidant component of CPS and it could be exploited as a valued antioxidant product.  相似文献   

18.
Three sulphated polysaccharides, coded as BEMPA, BEMPB1, BEMPB2, were extracted from the mucilage of mud snail of Bullacta exarata and purified by DEAE-cellulose ion-exchange and size-exclusion chromatography. Structural analysis of purified polysaccharides by chemical and biochemical methods revealed BEMPA was a high (1→3,4)-linked mannose-containing polysaccharide with molecular weight of 22,977 Da. BEMPB1, with molecular weight of 64,117 Da, was a high (1→3)-linked arabinose-containing polysaccharide. BEMPB2 was mainly composed of (1→3,4)-linked mannose with molecular weight of 47,507 Da. The comparison between sulphated polysaccharides and their desulphated products showed that sulphate substitutions of BEMPB1 were deduced to be at the C-3 of (1→4)-linked mannose, while sulphate substitutions of BEMPA and BEMPB2 were at C-4 of (1→3)-linked mannose. Furthermore, BEMPA exhibited highest inhibitory effects on growth of B-16 melanoma cells, and IC50 were 31.1 μg/mL.  相似文献   

19.
Maca (Lepidium meyenii Walp) polysaccharides (MP) with purity of 99.2% were obtained to investigate their structural characteristics and antifatigue effect in vivo. The physicochemical properties of MP were analyzed through high‐performance gel filtration chromatography, IR, monosaccharide composition, methylation, GC‐MS, and NMR analyses. The antifatigue effect of MP was evaluated by using a mouse weight‐loaded swimming model. MP is an acidic heteropolysaccharide with an average molecular weight (Mw) of 793.5 kDa. It is composed of D‐GalA: D‐Glc: L‐Ara: D‐Man: D‐Gal: L‐Rha = 35.07:29.98:16.98:13.01:4.21:0.75 (mol, %). The findings revealed that MP contained β‐1,3‐Galp(A), β‐1,3‐Glcp, and α‐1, 3‐Manp linked alternatingly to form a backbone (5:4:1). MP (above mid‐dosage 50 mg/kg bw/d) could effectively elongate swimming durations and accelerate average swimming speeds (within the 1st 5 min) of mice (P < 0.05) and improve the serous biochemical parameters of mice. Compared with the control model, high‐dosage (100 mg/kg bw/d) MP treatment could significantly enhance glutathione peroxidase and creatine kinase activities (P < 0.05) and decreased lactate dehydrogenase activity (P < 0.01). High‐dosage MP could significantly reduce the levels of blood urea nitrogen, lactic acid, and malondialdehyde (P < 0.05). MP is an acidic polysaccharide with a high D‐GalA content, which could be responsible for the antifatigue effect of maca.  相似文献   

20.
This study is focused on the isolation and characterisation of two purified polysaccharide fractions (namely PPEP‐1 and PPEP‐2) from Pleurotus eryngii (P. eryngii) and evaluation of their hypolipidaemic effects. The Congo red analysis indicated that PPEP‐2 but not PPEP‐1 possessed a triple‐helix conformation. The atomic force microscope analysis revealed that PPEP‐1 and PPEP‐2 showed different polysaccharide chain conformations. Importantly, the mice treated with PPEP‐1 showed significantly lowered serum levels of total cholesterol (TC), triglyceride (TG) and low‐density lipoprotein cholesterol (LDL‐c) and increased high‐density lipoprotein cholesterol when compared with the hyperlipidaemia mice induced by the high‐fat diet. However, PPEP‐2 revealed less hypolipidaemic effect than PPEP‐1. Additionally, both PPEP‐1 and PPEP‐2 demonstrated remarkable hypolipidaemic effects by decreasing levels of serum TC, TG and LDL‐c in the hyperlipidaemic model induced by P‐407. Taken together, our findings suggest that the P. eryngii polysaccharides, especially PPEP‐1, could be developed as a natural functional food supplement for preventing hyperlipidaemia.  相似文献   

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