Saccharide Mixture Research Articles

Behavioral evidence of saccharide mixture as a potent feeding effector for the purple mud crab Scylla tranquebarica

Behavioral evidence of saccharide mixture as a potent feeding effector for the purple mud crab Scylla tranquebarica

A novel maltooligosaccharide-forming α-amylase from Bacillus cereus and its application in the preparation of maltopentaose product

Maltooligosaccharides (MOS) have become a prominent market in the pharma and food industries due to their excellent bio-preservative and prebiotic properties. However, producing specific maltooligosaccharides like maltopentaose (G5) is relatively tedious and expensive. Here, a novel maltooligosaccharide-forming α-amylase from Bacillus cereus (BcMFAse) was characterized and exhibited a specific activity of 1965.0 ± 9.7 U/mg. The BcMFAse principally produced G5 by hydrolyzing soluble starch, with a maximum G5 yield of 215.8 ± 5.9 g/kg starch, accounting for 63.8 % of the saccharide mixture. The enzymatic hydrolysate was purified by AB-8 macroporous resin column chromatography. Briefly, 1.0 mL adsorbate containing 486.6 g/L total saccharides with deionized water elution at a flow rate of 0.7 mL/min and column temperature of 55 °C. Afterward, G5 was enriched up to 95.4 % purity, and 72.2 % was recovered in solution. The proposed process was simulated by SuperPro Designer software, and the remarkable economic results included a payback period of 0.08 years and an internal rate of return of 294.6 %. This study represented that BcMFAse could be a promising candidate for applications in the food and pharma fields. It first provided a high-efficiency process for producing > 90 % pure G5, which was economically attractive on an industrial scale.

No-additive salting-out liquid-liquid extraction-A tool for purification of positively charged compounds from highly salted reaction mixtures.

A novel and original application of salting-out assisted liquid-liquid extraction is presented. This technique was used to purify the final reaction products (quaternary ammonium salts) from unreacted components and by-products present in multiple excesses. The partition of two structurally related compounds as (2-aminoethyl)trimethylammonium salt (a labeling reagent) and a derivative of [2-(imidazoline-1-yl)ethyl]trimethylammonium salt (a final reaction product of N-acetylglucosamine labeling by (2-aminoethyl)trimethylammonium salt) between acetonitrile-rich and water-rich layers was monitored by hydrophilic interaction chromatography with electrospray ionization mass spectrometry. Despite the poor solubility of both highly polar substances in solutions containing a high concentration of acetonitrile, the main portion of the labeling reagent (72%) can be removed from the crude reaction mixture in the first extraction step using 95% acetonitrile/5% water as an extraction solvent. The purified final reaction product contained only 2% of the labeling reagent, and it was suitable for analysis by direct infusion mass spectrometry to confirm its identity. The capability of the suggested purification protocol to process small-volume highly salted reaction mixtures was also proven by analysis of saccharide mixture containing glucose, maltose, and maltotriose labeled by the positively charged tag.

Assessment of agave fructans as lyoprotectants of bovine plasma proteins concentrated by ultrafiltration

The effectiveness of fructans as protectant agent to prevent protein denaturation was evaluated during freeze-drying and storage. Native agave fructans and two fractions obtained by ultrafiltration were assessed as protective agents. Since the protein denaturation during freeze-drying can alter the functional properties of proteins, the evaluation of bovine plasma protein properties was performed with different concentration of fructans, pH and freezing temperatures. The results showed that the incorporation of fructans as lyoprotectants improved functional properties, due to the reduction of protein denaturation with maximum stabilization of plasma bovine protein at a fructan concentration between 5% and 10% (w/v) for native fructans and 10% (w/v) for the fractions of fructans at −40°C. Moreover, when freezing at −4°C, fructans fraction at 15% (w/v) prevented protein denaturation while native fructans exerted no acceptable protection (P<0.001). This behavior could be explained by a microstructure study of fructans, where the higher heterogeneity in size of native fructans may be the reason for the lower protective effect. Furthermore, the higher heterogeneity of the samples affected the extent of non-enzymatic browning in the range of the temperatures assessed. The shelf life of freeze-dried proteins was improved from 1.7months for the control sample to 7–11, depending on the saccharide considered. Taking into account the health benefits of fructans, since they are categorized as prebiotic, the protein–saccharide mixture may be valuable functional ingredients for food formulations.

Selective ethanol production from reducing sugars in a saccharide mixture

Fermentation profiles of four different yeasts reportedly defective in sucrose utilization indicate that all strains tested removed particular sugar via selective conversion to ethanol in a saccharide mixture. At the temperature of pressed sugarcane juice, Saccharomyces dairenensis and Saccharomyces transvaalensis performed better in ethanol production rate and yield, respectively.

Impact of saccharides and amino acids on the interaction of apple polyphenols with ion exchange and adsorbent resins

Model solutions containing catechin, caffeic acid, chlorogenic acid, phloridzin and rutin were used to examine their recovery using two adsorbent resins, four cation and four anion exchange resins at two temperatures (20 °C and 40 °C) and two pH values (pH 1 and 7). In addition, the interference of glucose, fructose and sucrose and of a saccharide mixture on polyphenol binding by one anion exchange and two adsorbent resins was assessed to approximate real conditions. In a further step polyphenol recovery using the aforementioned adsorbent and ion exchange resins was studied upon the addition of the amino acids alanine, glutamic acid, phenylalanine, lysine and histidine. The data reveal that both saccharides and amino acids influence polyphenol recovery positively and negatively, depending on the experimental conditions and the resin type; possible explanations for these findings are discussed. The information obtained in this study helps to gain a better understanding of the complex mechanisms of ion exchange and adsorption processes involving multi-component systems. The insights gained allow to design cost-efficient recovery of plant secondary metabolites from crude extracts for producing tailor-made health-beneficial fractions.

Preparation of 1-(2-naphthyl)-3-methyl-5-pyrazolone as pre-column derivatization reagent for separation and determination of saccharides using high performance liquid chromatography-mass spectrometry

The background and main task of the system of TCM fingerprintology from holism, systematology and complexity science are elaborately discussed. The fingerprintology of TCM is a novel system, in which the pharmacologic substance bases, mechanism and the law of pharmacokinetics on TCM (herbal medicine) and technologies for other related preparations are systematically and integrally studies. The core position and bridge effect of the fingerprint informatics of TCM were set forth. The system of TCM fingerprintology comprises the fingerprint testology, the fingerprint quality controlology, the fingerprint pharmacodynamics, the fingerprint pharmacokinetic, the fingerprint-pharmaceutics and the biofingerprintology of TCM. The theories and methods of complexity science and system science should be applied in the study of the system. The new mode of the innovative development of TCM is established by breaking through the linear thinking and reduction analysis, highly emphasizing mathematical principles and methods from a integral and systematic point of view. The theories and methods of the TCM fingerprintology are the main forces in the analysis of the leading technology and modernization of TCM. The maturity and perfection of the theories and methods will powerfully support he development of innovative TCM. The ultimate purpose of the system of the TCM fingerprintology is to develope effective, safety and controllable TCMs for human and other beneficial organisms.

Photoinduced biohydrogen production from saccharide mixture with the photosensitization of Mg chlorophyll a from green plant

Abstract Here, we developed a photoinduced biohydrogen production system coupling saccharide hydrolysis with hydrogen production. A mixture of three different saccharides (sucrose, maltose, and cellobiose) was hydrolyzed by three enzymes (invertase, glucoamylase, and cellulase) and glucose dehydrogenase (GDH). Hydrogen production was performed with colloidal platinum as a catalyst, using visible light-induced photosensitization of Mg chlorophyll a (Mg Chl a ). Irradiation of the sample solution containing the saccharides, enzymes, GDH, NAD + , Mg Chl a , methyl viologen (MV 2+ ), and colloidal platinum resulted in continuous hydrogen production. Approximately 2.9 μmol hydrogen was produced after 2 h of irradiation, and the total yield was approximately 8.8%.

Fluorophore-assisted carbohydrate electrophoresis as detection method for carbohydrate-protein interactions

Fluorophore-assisted carbohydrate electrophoresis (FACE) is a straightforward, sensitive method for determining the presence and relative abundance of individual (oligo)saccharide in a(n) (oligo)saccharide mixture. The single terminal aldehydes of (oligo)saccharides were tagged with the charged fluorophore 8-aminonaphthalene-1,3,6-trisulfonate (ANTS), and separated with high resolution on the basis of size by polyacrylamide gel electrophoresis. ANTS fluorescence labeling is not biased by (oligo)saccharide length. Therefore, band fluorescence intensity is directly related to the relative abundance of individual (oligo)saccharide moieties in heterogeneous sample. In the same time, it also indicates that FACE can be used to investigate the interactions of carbohydrates and proteins.

The analysis of oligosaccharides derived from different sources by fluorophore-assisted carbohydrate electrophoresis

Fluorophore-assisted carbohydrate electrophoresis (FACE) is a straightforward, sensitive method for determining the presence and relative abundance of individual (oligo)saccharide in a(n) (oligo)saccharide mixture. The single terminal aldehydes of oligoglucoside residues released by acid hydrolysis of β-1, 3- d-glucan from yeast were tagged with the charged fluorophore 8-aminonaphthalene-1,3,6-trisulfonate (ANTS), and separated with high resolution on the basis of size by polyacrylamide gel electrophoresis. ANTS fluorescence labeling was not biased by oligoglucoside length; therefore, band fluorescence intensity was directly related to the relative abundance of individual oligoglucoside moieties in heterogeneous sample. Therefore, FACE represents an accessible, sensitive, and quantitative analytical tool enabling the analysis of (oligo)saccharides derived from different sources.

Hydrolysis characteristics of a β‐1,3‐d‐glucan elicitor from yeast

Fluorophore-assisted carbohydrate electrophoresis (FACE) is a straightforward, sensitive method for determining the presence and relative abundance of individual (oligo)saccharides in a(n) (oligo)saccharide mixture. The single-terminal aldehydes of oligoglucoside residues released by acid hydrolysis of beta-1,3-D-glucan from yeast were tagged with the charged fluorophore ANTS (8-aminonaphthalene-1,3,6-trisulphonate), and separated with high resolution on the basis of size by PAGE. ANTS fluorescence labelling was not biased by oligoglucoside length; therefore band fluorescence intensity was directly related to the relative abundance of individual oligoglucoside moieties in a heterogeneous sample. FACE analysis revealed that the major oligoglucoside mixture released by acid hydrolysis from beta-1,3-D-glucan was composed of monosaccharide, disaccharide, trisaccharide, tetrasaccharide, pentasaccharide, hexasaccharide, heptasaccharide and octasaccharide, and the order of abundance from high to low was trisaccharide, monosaccharide, disaccharide, tetrasaccharide, pentasaccharide, hexasaccharide, heptasaccharide and octasaccharide respectively. In conclusion, FACE represents an accessible, sensitive and quantitative analytical tool enabling the characterization of a(n) (oligo)saccharide mixture.

Studies on the Hydrolytic Condition of β‐1,3 Glucan from Yeast by Fluorophore‐Assisted Carbohydrate Electrophoresis

Fluorophore‐assisted carbohydrate electrophoresis (FACE) is a straightforward, sensitive method for determining the presence and relative abundance of individual (oligo)saccharide in a(n) (oligo)saccharide mixture. The single‐terminal aldehydes of oligoglucoside residues released by acid hydrolysis of β‐1,3 glucan from yeast were tagged with the charged fluorophore 8‐aminonaphthalene‐1,3,6‐trisulfonate (ANTS) and separated with high resolution on the basis of size by polyacrylamide gel electrophoresis. ANTS fluorescence labeling was not biased by oligoglucoside length; therefore, band fluorescence intensity was directly related to the relative abundance of individual oligoglucoside moieties in heterogeneous samples. FACE analysis revealed that the major oligoglucoside mixture released by acid hydrolysis from β‐1,3 glucan was composed of monosaccharide, disaccharide, trisaccharide, and tetrasaccharide, respectively. And the best hydrolytic condition is at a concentration of 2.0 mol/L TFA, heated at 85°C for 2.0 h. In conclusion, FACE represents an accessible, sensitive, and quantitative analytical tool enabling the characterization of a(n) (oligo)saccharide mixture. The results of these bioassays demonstrated that the sample obtained on the condition of 2.0 h, 85°C, 2.0 mol/L was the most active as compared to the sample obtained on the other conditions in eliciting phytoalexin accumulation in tobacco cotyledon tissue. The project was financially supported by “The Scaling the Height Program” of the State Science and Technology Committee of China.

Intravenous synthetic alphaGal saccharides delay hyperacute rejection following pig-to-baboon heart transplantation.

Several oligosaccharides containing the terminal structure Gal(alpha)1-3Gal (alphaGal) and different side chains were tested in vitro for their ability to block natural anti(alpha)Gal antibodies. A di-and a trisaccharide (di(alpha)Gal and tri(alpha)Gal) were selected. A blood group B baboon, having IgG and IgM natural antipig titers of 1:256 and 1:1024 and a hemolytic titer (to pig red blood cells, RBCs) of 1:8, was chosen to measure pharmacokinetic parameters of the saccharides and to assess the extent of in vivo neutralization of the antibodies. Three grams each of the di(alpha)Gal and the tri(alpha)Gal dissolved in saline were administered by bolus intravenous (i.v.) injection. Blood samples were collected at various times and urine was collected at 8 and 24 h. Plasma and urine concentrations of the alphaGal saccharides were estimated by an ELISA specially developed for this study. A fast distribution phase followed by equilibrium and excretion phases were observed, indicating a T1/2 in the order of 1 h. Fifty-eight per cent of the saccharides were recovered in the urine within 24 h. Determination of antipig antibody binding by FACS analysis and of serum cytotoxicity titers for pig endothelial cells demonstrated that a 70% reduction in binding and cytotoxicity could be achieved with plasma saccharide levels of 300-400 microg/ml. Six months later, a pig heart was transplanted heterotopically into the baboon. A 3-g bolus of the saccharide mixture (1.5 g of each saccharide) was given i.v. before allowing blood reperfusion of the transplanted heart, followed by an i.v. infusion of 1 g/hr for 1 hr and 0.5 g/hr for the 3 succeeding hours. Blood concentrations of the saccharides, CH50, hematology and cytotoxicity for PK15 cells were estimated in blood samples taken at various times. Heart function was observed to be satisfactory for 8 h, but was found to have ceased at 18 h. Myocardial biopsies taken at 3 and 5 h showed congestion only, suggestive of minimal vascular rejection, but by 18 h demonstrated severe vascular rejection. In conclusion, alphaGal saccharide therapy given for a period of 4 h delayed, but did not totally prevent, the development of vascular rejection in the pig-to-baboon heart transplant model. alphaGal saccharide therapy may be one of several useful approaches for the prevention of hyperacute rejection in pig-to-primate organ transplantation.

Formation of Fine Oil-in-Water Emulsions by the Gel Emulsification Method with Saccharide and Protein

An emulsification method using a gel-like phase of a saccharide and protein mixture has been developed. In the method, which is called a gel emulsification method, an oil is added to the highly concentrated saccharide solution containing protein to form a clear gel-like phase, which followed by dilution with water to form a fine oil-in-water emulsion. This emulsion was investigated as to its emulsifying activity and emulsion stability as compared with that obtained by high-shear equipment, which was called a homomixer method. The emulsifying activity of the emulsions prepared by the gel emulsification method was much higher than that of the emulsions prepared by the homomixer method. The emulsions prepared by both methods were highly stable in terms of the stability against coalescence. On the other hand, the stability against creaming of the emulsions prepared by the gel emulsification method was much higher than that of the emulsions prepared by the homomixer method. The surface hydrophobicity of the protein and the unfreezable water content in the highly concentrated saccharide solution containing protein were not correlated to the emulsifying properties of the emulsions prepared by the gel emulsification method, which appeared to be dependent on the viscosity of the highly concentrated saccharide solution containing protein.

The high maltose-producing α-amylase of the thermophilic actinomycete, Thermomonospora curvata

The alpha-amylase of Thermomonospora curvata catalyses the formation of very high levels of maltose from starch (73%, w/w) without the attendant production of glucose. The enzyme was produced extracellularly in high yield during batch fermentation in a 5-1 fermentor. Purification was achieved by ammonium sulphate fractionation, Superose-12 gel filtration and DEAE-Sephacel ion-exchange chromatography. The enzyme exhibited maxima for activity at pH 6.0 and 65 degrees C, had a relative molecular mass of 60,900-62,000 and an isoelectric point at 6.2. The exceptionally high levels of maltose produced and the unique action pattern exhibited on starch and related substrates indicate a very unusual maltogenic system. The predominance of maltose as the final end-product may be explained by the participation of reactions other than simple hydrolysis and the preferential cleavage of maltotriose from higher maltooligosaccharides. The enzyme exhibits very low affinity for maltotriose (Km = 7.7 x 10(-3) M) and its conversion to maltose is achieved by synthetic followed by hydrolytic events, which result in the very high levels of maltose observed and preclude glucose formation. This system is distinguished from other very high maltose-producing amylases by virtue of its high temperature maximum, very low affinity for maltotriose and the absence of glucose in the final saccharide mixture.

Convenient, in-line purification of saccharide mixture in automated high- performance liquid chromatography

Convenient, in-line purification of saccharide mixture in automated high- performance liquid chromatography

An automated anion-exchange chromatographic procedure for the estimation of saccharides in acid hydrolysates of soil

Abstract It is well known that the major neutral monosaccharide components released from soil by acid hydrolysis are glucose, galactose, mannose, arabinose, xylose, ribose, rhamnose and fucose. A colorimetric determination of the saccharide mixture released from soil is unsatisfactory for determining an accurate figure for soil saccharides. More precise information can be obtained by the determination of monosaccharides after separation by chromatography. Paper and thin-layer chromatography for quantitative analysis are rather time consuming and laborious. The gas chromatographic procedure was applied successfully for the analysis of sugars in soil hydrolysates by OADES et at. (7). Preparation of the various derivatives for gas chromatography still requires many steps, much handling of the sample, and considerable time, although final analysis of the product derivative is accomplished in an hour or two.