scholarly journals Anti-coagulant Activity of Isolated and Partially Characterized Proteoglycans and Glycosaminoglycans from African Night Crawler (Eudrilus eugeniae Kinberg)

KIMIKA ◽  
2015 ◽  
Vol 26 (2) ◽  
pp. 31-38
Author(s):  
Mia Clare Marie L. Bercansil ◽  
Miko Lorenzo J. Belgado
Keyword(s):  
Hyaluronic Acid ◽  
Infrared Spectroscopy ◽  
Enzymatic Hydrolysis ◽  
Gel Filtration ◽  
Acetate Buffer ◽  
Eudrilus Eugeniae ◽  
Gel Filtration Chromatography ◽  
Molecular Weights ◽  
Coagulant Activity ◽  
Hydrolysis Of

Proteoglycans and glycosaminoglycans were isolated from African night crawler (Eudrilus eugeniae Kinberg) and partially characterized proteoglycans (3.04 % of lyophilized worm) were liberated from the defatted and depurinated worm samples by dissociative method using 4M urea in acetate buffer. Glycosaminoglycans (12.47% of proteoglycan extract) were extracted using enzymatic hydrolysis of the proteoglycan extract with papain. Gel filtration chromatography using Sepharose CL-4B was used to purify and estimate the molecular weights of the proteoglycan and glycosaminoglycan fractions. Three proteoglycan fractions PGF1, PGF2 and PGF3 with estimated molecular weigths 860 kDa, 181 kDa and 3 kDa, respectively were identified as monitored by the Bradford and modified carbazole assay. Two glycosaminoglycan fractions - GF1 (MW = 860 kDa) and GF2 (MW=140 kDa) were identified using the modified carbazole assay. Infrared spectroscopy of the GF1 and GF2 showed the possible identities of the fractions. GF1 may be a hyaluronic acid and GF2 is possibly chondroitin. Anti-coagulant assay for the extracts and fractions revealed that the glycosaminoglycan isolate has anti-coagulant activity but not the GF1 and GF2 fractions individually.

Enzymatic hydrolysis of chitosan in acetate buffer solution

2015 ◽  
Vol 88 (4) ◽  
pp. 647-651 ◽  
Author(s):  
V. V. Chernova ◽  
A. S. Shurshina ◽  
M. V. Bazunova ◽  
E. I. Kulish
Keyword(s):  
Enzymatic Hydrolysis ◽  
Acetate Buffer ◽  
Acetate Buffer Solution ◽  
Buffer Solution ◽  
Hydrolysis Of

scholarly journals Purification and Characterization of 2,6-β-d-Fructan 6-Levanbiohydrolase fromStreptomyces exfoliatus F3-2

2000 ◽  
Vol 66 (1) ◽  
pp. 252-256 ◽  
Author(s):  
Katsuichi Saito ◽  
Kazuya Kondo ◽  
Ichiro Kojima ◽  
Atsushi Yokota ◽  
Fusao Tomita
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Extracellular Enzyme ◽  
Molecular Weights ◽  
Sds Page ◽  
Monomeric Structure ◽  
Ph Range ◽  
Hydrolysis Of

ABSTRACT Streptomyces exfoliatus F3-2 produced an extracellular enzyme that converted levan, a β-2,6-linked fructan, into levanbiose. The enzyme was purified 50-fold from culture supernatant to give a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weights of this enzyme were 54,000 by SDS-PAGE and 60,000 by gel filtration, suggesting the monomeric structure of the enzyme. The isoelectric point of the enzyme was determined to be 4.7. The optimal pH and temperature of the enzyme for levan degradation were pH 5.5 and 60°C, respectively. The enzyme was stable in the pH range 3.5 to 8.0 and also up to 50°C. The enzyme gave levanbiose as a major degradation product from levan in an exo-acting manner. It was also found that this enzyme catalyzed hydrolysis of such fructooligosaccharides as 1-kestose, nystose, and 1-fructosylnystose by liberating fructose. Thus, this enzyme appeared to hydrolyze not only β-2,6-linkage of levan, but also β-2,1-linkage of fructooligosaccharides. From these data, the enzyme from S. exfoliatus F3-2 was identified as a novel 2,6-β-d-fructan 6-levanbiohydrolase (EC 3.2.1.64 ).

scholarly journals Characterization of Melanoidins and Color Development in Dulce de Leche, a Confectionary Dairy Product With High Sucrose Content: Evaluation of pH Effect, an Essential Manufacturing Process Parameter

2021 ◽  
Vol 8 ◽  
Author(s):  
Analía Rodríguez ◽  
Patricia Lema ◽  
María Inés Bessio ◽  
Guillermo Moyna ◽  
Cristina Olivaro ◽  
...  
Keyword(s):  
Enzymatic Hydrolysis ◽  
Ph Effect ◽  
Dairy Product ◽  
Gel Filtration ◽  
Significant Degree ◽  
Product Analysis ◽  
Molecular Weights ◽  
Ph Values ◽  
Initial Ph

The effect on color of the initial pH employed in dulce de leche (DL) production was evaluated through physicochemical and spectroscopical characterization of the melanoidins formed in the process. Melanoidins originated at pH values of 6.5, 7.0, and 7.5, and they were released by the enzymatic hydrolysis of the protein backbone and purified by gel filtration. They showed a significant degree of polydispersity, in general, with molecular weights (MWs) below 1,800 Da. DL produced at a higher pH released melanoidins with higher average MW after the enzymatic hydrolysis. They also presented darker colors (dE*ab, C*), more closely resembling those typical of the commercial product. Analysis of the fractions isolated by gel filtration using HPLC-DAD and multinuclear NMR showed an heterogeneous and complex composition. Even though structurally related, the 1H NMR spectra of melanoidins showed a higher degree of aromaticity at higher pH values. In conclusion, the pH employed in DL production affects the amount and structure of the colored products originated by MR reactions, and thus the color of the final product.

Plasma Met-enkephalin and catecholamine responses to insulin-induced hypoglycaemia in greyhounds

1987 ◽  
Vol 114 (1) ◽  
pp. 81-87 ◽  
Author(s):  
S. Medbak ◽  
D. F. J. Mason ◽  
L. H. Rees
Keyword(s):  
Blood Glucose ◽  
Plasma Concentrations ◽  
Gel Filtration ◽  
Opioid Antagonist ◽  
Molecular Forms ◽  
Endogenous Opioid ◽  
Gel Filtration Chromatography ◽  
Plasma Adrenaline ◽  
Molecular Weights ◽  
Blood Glucose Concentrations

ABSTRACT The involvement of endogenous opioid peptides in the stress response was investigated by measuring plasma concentrations of Met-enkephalin-like immunoreactivity (MLI), adrenaline and noradrenaline during insulin-induced hypoglycaemia in conscious greyhounds. Moreover, the molecular forms of circulating MLI were characterized using gel filtration chromatography. In the first group of animals, i.v. administration of insulin (0·3 units/kg) provoked marked hypoglycaemia (blood glucose concentrations fell from 4·4 ± 0·1 to 1·5 ±0·2 mmol/l; mean ± s.e.m.) which was associated with significant (P< 0·001) rises in plasma MLI concentrations from a basal concentration of 45 ± 8 to a peak of 189 ±39 ng/l. A within-subject study comparing five different insulin doses ranging from 0·004 to 0·3 units/kg showed dose-related effects on blood glucose with nadir concentrations of 4·1 ± 0·6 mmol/l (after the smallest dose of insulin) and 0·8 ± 0·1 mmol/l (after the largest dose of insulin). This was associated with dose-related rises in plasma MLI with peak concentrations of 56±17 and 558 ± 35 ng/l, plasma adrenaline with peak concentrations of 0·45± 0·06 and 15·76±1·33 nmol/l and plasma noradrenaline with peak concentrations of 0·49 ± 0·07 and 2·27 ± 0·45 nmol/l following the smallest and largest doses of insulin respectively. These results are the first demonstration of raised plasma MLI concentrations following hypoglycaemia. Moreover, they show that the hormonal responses vary with the degree of hypoglycaemia achieved. Together with reports by other investigators these findings might suggest opioid modulation of the responses of the sympathoadrenal system to hypoglycaemia. These responses were, however, not modified by the opioid antagonist naloxone. Gel filtration chromatography of neat (unextracted) plasma revealed the predominance of large molecular weight enkephalin-containing peptides, with approximate molecular weights of 18 000 and 8000, both basally and following stimulation by hypoglycaemia. J. Endocr. (1987) 114,81–87

scholarly journals Comparison of Enzymatic Hydrolysis of Polysaccharides from Eggshells Membranes

2016 ◽  
Vol 15 (2) ◽  
pp. 133-141 ◽  
Author(s):  
Eva Ürgeová ◽  
Katarína Vulganová
Keyword(s):  
Extracellular Matrix ◽  
Hyaluronic Acid ◽  
Enzymatic Hydrolysis ◽  
Glucuronic Acid ◽  
Optimal Conditions ◽  
Osteoarthritis Treatment ◽  
Neural Tissues ◽  
Eggshell Membranes ◽  
Hydrolysis Of ◽  
And Cartilage

Abstract AHyaluronic acid (HA) is part of the extracellular matrix of connective, epithelial and neural tissues, as well as the synovial fluid, skin, and cartilage. It is composed of repeating disaccharide units of D-glucuronic acid and N-acetyl glucosamine. Hyaluronic acid is used in abdominal surgery, ophthalmology, dermatology, rhinology; it is usable for the osteoarthritis treatment. The membranes of eggshell are a natural source of hyaluronic acid, collagen, glycosaminoglycan and collagenous proteins. In paper, we tested the possibility of extraction hyaluronic acid from the eggshell membranes by enzymatic hydrolysis. We identified optimal conditions of hydrolysis with trypsin at reaction temperature of 37 °C and pH 8; with pepsin at 40 °C and pH 3, as well as with papain at 60 °C and pH 7.5. The content of hyaluronic acid in samples was determined spectrophotometrically using the carbazole method. The experimental results showed a yield of ~ 4 -4.5 % hyaluronic acid per 1 g of dry eggshell membranes.

scholarly journals Tumour calcitonin. Interaction with specific calcitonin receptors

1980 ◽  
Vol 190 (3) ◽  
pp. 545-550 ◽  
Author(s):  
J Ham ◽  
M L Ellison ◽  
J Lumsden
Keyword(s):  
Thyroid Carcinoma ◽  
Cell Line ◽  
Medullary Thyroid Carcinoma ◽  
Gel Filtration ◽  
Bronchial Carcinoma ◽  
Binding Activity ◽  
Gel Filtration Chromatography ◽  
Medullary Thyroid ◽  
Molecular Weights ◽  
Receptor Binding Activity

The human epidermoid bronchial carcinoma (BEN) cell line has been shown to have specific membrane binding sites for calcitonin and to secrete high-molecular-weight forms (ranging from 40000 to 10000) of immunoreactive calcitonin. Synthetic salmon and human calcitonins and a thyroid extract of porcine calcitonin have been shown to displace 125I-labelled salmon calcitonin from the receptors in a dose-related fashion. The binding to these receptors of calcitonins derived from the BEN cell line and a medullary thyroid carcinoma with molecular weights ranging from 28000 to 3500 (both separated by gel-filtration chromatography) has been investigated. Neither major peaks of BEN-cell-line calcitonin showed receptor binding activity. Only one form of medullary thyroid carcinoma calcitonin, that which co-eluted with synthetic calcitonin monomer on gel-filtration chromatography, caused any significant displacement of labelled hormone from the receptors.

Two types of xylanases of alkalophilic Bacillus sp. No. C-125

1985 ◽  
Vol 31 (6) ◽  
pp. 538-542 ◽  
Author(s):  
H. Honda ◽  
T. Kudo ◽  
Y. Ikura ◽  
K. Horikoshi
Keyword(s):  
Ammonium Sulfate ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Bacillus Sp ◽  
Ammonium Sulfate Precipitation ◽  
Molecular Weights ◽  
Protein Molecules ◽  
Activity Curve ◽  
Alkalophilic Bacillus ◽  
Hydrolysis Of

One alkalophilic Bacillus sp. strain C-125 (FERM No. 7344) was isolated from soil. From this organism, two types of xylanases, designated xylanase A and xylanase N, were purified by an ammonium sulfate precipitation followed by Biogel P-30 gel filtration, DEAE-cellulose chromatography, and Sephadex G-75 gel filtration. The molecular weights of xylanase A and N were estimated as 43 000 and 16 000, respectively. Immunological experiments indicated that xylanase A and xylanase N were entirely different protein molecules. Xylanase N was most active at pH 6.0–7.0, but xylanase A had a very broad pH activity curve (pH 6–10) and was still active even at pH 12.0. The maximum hydrolysis of xylan by the enzymes was about 25%. Both enzymes split xylan and yielded xylobiose and higher oligosaccharides but could hydrolyze neither xylobiose nor xylotriose. Trans xylosidation activities were detected in both enzymes.

scholarly journals Improvement of the method of obtaining human IgA Fc-fragments

2015 ◽  
Vol 6 (1) ◽  
pp. 32-35
Author(s):  
O. Y. Galkin ◽  
Y. V. Gorshunov ◽  
V. F. Solovjova
Keyword(s):  
Enzymatic Hydrolysis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Inhibitory Effect ◽  
Nitrogen Atmosphere ◽  
Maximum Output ◽  
Serological Tests ◽  
Isolation And Purification ◽  
Antibody Conjugate ◽  
Hydrolysis Of

To address a number of fundamental and applied problems in immunology, molecular and cellular biology and biotechnology it is necessary to obtain Fc-fragments of immunoglobulins. Fc-fragments may be used for studying of the effector functions of antibodies which are mediated by these areas. They are often used as an immunogen to produce anti-specie (based on so-called secondary antibody) conjugate in the development of serological tests for diagnostics (predominantly such conjugate based on monoclonal antibodies). The work is aimed to develop improved methods of obtaining and allocation of Fc-fragments of human IgA. To achieve this objective, optimization of hydrolysis of IgA with subsequent purification of Fс-fragments have been carried out. Improved method of obtaining Fc-fragments of IgA provides: papain hydrolysis of immunoglobulin in the environment of nitrogen for 4 h, allowing to achieve maximum output of Fc-fragments without their further degradation: isolation and purification of Fc-fragments of human IgA by one-stage gel filtration on sephadex G-100; control of purity of the target product by electrophoresis in polyacrylamide gel with sodium dodecyl sulfate and Ouchterlony immunodiffusion. Enzymatic hydrolysis was carried out at the optimal temperature of papain (37 °C). As the oxygen in the air may have inhibitory effect on enzymatic hydrolysis reaction, the reaction mixture was incubated in the nitrogen atmosphere to prevent inactivation of papain. To reduce the incident degradation of immunoglobulin molecules, papain hydrolysis was carried out without using an enzyme activator (cysteine). Usage of the proposed scheme allows obtaining Fc-fragments of human IgA of high purity. Outcome of Fc-fragments after all stages of purification was about 18% of the initial amount of IgA in the preparation. Molecular weight from Fc-fragments of human IgA was equal to approximately 70 kDa.

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