scholarly journals Identification and partial purification of thermally stable peroxidase isoenzymes from seedlings of Vigna sp. (V) landrace Vn

2021 ◽  
Vol 12 (3) ◽  
pp. 476-483
Author(s):  
Yves Mann Elate Lea Mbassi ◽  
Marie Solange Evehe ◽  
Wilfred Fon Mbacham ◽  
John Payne Muluh
Keyword(s):  
Thermal Stability ◽  
Deae Cellulose ◽  
Exchange Chromatography ◽  
Partial Purification ◽  
Heat Inactivation ◽  
Inactivation Kinetics ◽  
Potential Candidate ◽  
Peroxidase Isoenzymes ◽  
The North ◽  
Cold Acetone

Several soluble peroxidase isoenzymes are expressed in a landrace of Vigna sp. cultivated in the north of Cameroon (landrace called Vn in previous study) during seed germination. There are at least two cathodic peroxidases and eight major anodic peroxidases as shown by their electrophoretic migration at pH 7.4 under native conditions. These isoperoxidases are more expressed in roots than in shoots. They have different thermal stability, so that heat inactivation kinetics of crude peroxidase extracts from roots do not fit the first-order model. The slow and intermediate migrating groups of anodic isoperoxidases retains a substantial activity after ten minutes of incubation at 80°C and 85°C. An anodic isoperoxidase (named A6 in this study) shows in addition to this great thermal stability, a high activity in seedlings and is expressed both in roots and shoots. The combination of those characteristics makes this isoperoxidase a potential candidate for biotechnological applications. Three major anodic isoperoxidases, of which A6 and another thermostable isoperoxidase, were successfully separated from each other by ion exchange chromatography on DEAE-cellulose, after precipitation of total proteins by ice-cold acetone. This offers the prospect of being able to characterize these isoperoxidases individually in future studies.

scholarly journals Peroxidase isoenzymes from seedlings of VIGNA sp (V) landrace Vn: distribution and thermal stability

2021 ◽  
Author(s):  
Yves Mann Elate Lea Mbassi ◽  
Marie Solange Evehe Bebandoue ◽  
Wilfred Fon Mbacham
Keyword(s):  
Thermal Stability ◽  
Heat Inactivation ◽  
Inactivation Kinetics ◽  
Potential Candidate ◽  
Order Model ◽  
Peroxidase Isoenzymes ◽  
First Order ◽  
The North ◽  
Soluble Peroxidase ◽  
Kinetics Of

Abstract Several soluble peroxidase isoenzymes are expressed in a landrace of Vigna sp cultivated in the north of Cameroon (landrace called Vn in previous study) during seed germination. There are at least two cathodic peroxidases and eight major anodic peroxidases as shown by their electrophoretic migration at pH 7.4 under native conditions. These isoperoxidases are more expressed in radicles than in shoots. They have different thermal stability, so that heat inactivation kinetics of crude peroxidase extracts from radicles does not fit the first-order model. One major anodic isoperoxidase of the slow migrating group and at least two others anodic isoperoxidases of the The slow and intermediate migrating groups of anodic isoperoxidasesanodic isoperoxidases are stable for ten minutes of incubation at 80°C and 85°C. The major anodic isoperoxidase of the The less anodic slow migrating groupisoperoxidase (named A6 in this study) shows in addition to this great thermal stability, a high activity during germination and is expressed both in radicles and shoots in large amounts. The combination of those characteristics makes thisthat isoperoxidase a a potential candidate for biotechnological applications.

scholarly journals The partial purification and characterization of cytosol alcohol dehydrogenase from Astasia

1974 ◽  
Vol 141 (2) ◽  
pp. 469-475 ◽  
Author(s):  
Rolf Morosoli ◽  
Nicole Bégin-Heick
Keyword(s):  
Molecular Weight ◽  
Alcohol Dehydrogenase ◽  
Polyacrylamide Gel Electrophoresis ◽  
Deae Cellulose ◽  
Total Activity ◽  
Growth Conditions ◽  
Partial Purification ◽  
Heat Inactivation ◽  
Kinetic Properties ◽  
Ph Optimum

1. The cytosol alcohol dehydrogenase (alcohol–NAD oxidoreductase, EC 1.1.1.1) of Astasia longa was partially purified and characterized from cells grown in the presence of air+CO2 (95:5) or of O2+CO2 (95:5). 2. Under both these growth conditions, the cells contained a fraction, ADHII, which was characterized by its electrophoretic properties, by a high degree of resistance to heat inactivation, by a sharp pH optimum at 8.2 and by its kinetic properties. The estimated molecular weight of this fraction was approx. 150000, which is similar to that of yeast alcohol dehydrogenase. 3. Cells grown in air+CO2 (95:5) contain another fraction, ADHI, which can be further separated into two subfractions by polyacrylamide-gel electrophoresis and by DEAE-cellulose chromatography. This was termed fraction ‘ADHI-air’. 4. In addition to fraction ADHII, cells grown in the presence of O2 have a twofold increase in fraction ADHI-air activity as well as two new fractions that could not be demonstrated in air-grown cells. These new fractions which we have called fraction ‘ADHI-O2’, account for about 10% of the total activity. 5. The ADHI fractions (air) and (O2) have similar broad pH–activity curves and similar kinetic properties, both having a lower Km for ethanol and NAD than fraction ADHII. However, they differ from each other with respect to their activity with various substrates. The estimated molecular weight of these two ADHI fractions and their chromatographic behaviour on hydroxyapatite and on DEAE-cellulose also distinguish them.

scholarly journals Heat inactivation kinetics of Hypocrea orientalis β-glucosidase with enhanced thermal stability by glucose

2015 ◽  
Vol 81 ◽  
pp. 1012-1018 ◽  
Author(s):  
Xin-Qi Xu ◽  
Yan Shi ◽  
Xiao-Bing Wu ◽  
Xi-Lan Zhan ◽  
Han-Tao Zhou ◽  
...  
Keyword(s):  
Thermal Stability ◽  
Heat Inactivation ◽  
Inactivation Kinetics ◽  
Kinetics Of

scholarly journals N-acetyl-β-d-glucosaminidase in marmoset kidney, serum and urine

1978 ◽  
Vol 175 (3) ◽  
pp. 859-867 ◽  
Author(s):  
R J Pierce ◽  
R G Price ◽  
J S L Fowler
Keyword(s):  
Gel Electrophoresis ◽  
Human Liver ◽  
Deae Cellulose ◽  
Heat Stability ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Partial Purification ◽  
Starch Gel Electrophoresis ◽  
Starch Gel ◽  
Serum And Urine

N-Acetyl-beta-D-glucosaminidase activities were determined in homogenates of marmoset kidney, in serum and in urine by using the 4-methylumbelliferyl substrate. The enzyme activity was separated into several components by DEAE-cellulose ion-exchange chromatography, starch-gel electrophoresis and isoelectric focusing. The kidney contained two major forms of the enzyme, A and B, which had similar pH optima and Km values. The A-form bound to DEAE-cellulose at pH 6.8, migrated towards the anode on starch-gel electrophoresis and had a pI of 5.0. The B-form did not bind to DEAE-cellulose at pH 6.8, remained near the origin on starch-gel electrophoresis and had a pI of 7.64. The isoenzymes also differed in heat stability, the B-form being the more stable. Serum contained B-form activity and, in addition, two intermediate forms (I1 and I2) were loosely bound to DEAE-cellulose. The serum A-form activity was less firmly bound to DEAE-cellulose than was the tissue A-form and was designated As. Serum from a pregnant marmoset contained a form which may be analogous to the human P-isoenzyme. Urine contained only a small amount of B-form activity, the majority being present in the A-form. The kidney A- and B-forms both had mol.wts. of 96000–100000 and the activity was predominantly lysosomal. Partial purification of the kidney A isoenzyme was undertaken. Immunoprecipitation studies indicated a relationship between marmoset kidney A-form and human liver A-form activity.

BETA-D-GLUCANASES OF SCHIZOPHYLLUM COMMUNE

1967 ◽  
Vol 13 (8) ◽  
pp. 969-978
Author(s):  
G. L. Schneberger ◽  
W. W. Luchsinger
Keyword(s):  
Ion Exchange ◽  
Schizophyllum Commune ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Heat Inactivation ◽  
Inactivation Kinetics ◽  
Hydrolysis Products ◽  
Culture Filtrates

Culture filtrates of Schizophyllum commune have been subjected to ion-exchange chromatography, and the laminarin-splitting and endo-β-glucanase activities have been studied with respect to pH behavior, heat inactivation kinetics, effects of inhibitors, and hydrolysis products. Evidence is presented to show that the two activities may be due to the same enzyme and that a β-(1 → 3) linkage may be involved in the specificity of the enzyme.

scholarly journals Extraction and Partial Purification of Mouse Uterine Alkaline Phosphatase

1979 ◽  
Vol 32 (2) ◽  
pp. 153 ◽  
Author(s):  
RN Murdoch ◽  
DJ Kay ◽  
WJ Capper
Keyword(s):  
Alkaline Phosphatase ◽  
Ammonium Sulfate ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Partial Purification ◽  
Ammonium Sulfate Precipitation ◽  
Pregnant Mice ◽  
Triton X

Alkaline phosphatase in uterine homogenates from day 7 pregnant mice was solubilized using 0�2 % (v/v) Triton X-100 and extracted with 20% (v/v) n-butanol. The procedure, which resulted in 182- fold purification, included ammonium sulfate precipitation, DEAE-cellulose anion exchange chromatography and Sephadex 0200 gel filtration.

Studies on an Inhibitor of Plasminogen Activation in Human Serum

1973 ◽  
Vol 30 (02) ◽  
pp. 414-424 ◽  
Author(s):  
Ulla Hedner
Keyword(s):  
Ion Exchange ◽  
Human Serum ◽  
Antithrombin Iii ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Specific Adsorption ◽  
Partial Purification ◽  
Plasminogen Activation ◽  
Preparative Electrophoresis

SummaryA procedure is described for partial purification of an inhibitor of the activation of plasminogen by urokinase and streptokinase. The method involves specific adsorption of contammants, ion-exchange chromatography on DEAE-Sephadex, gel filtration on Sephadex G-200 and preparative electrophoresis. The inhibitor fraction contained no antiplasmin, no plasminogen, no α1-antitrypsin, no antithrombin-III and was shown not to be α2 M or inter-α-inhibitor. It contained traces of prothrombin and cerulo-plasmin. An antiserum against the inhibitor fraction capable of neutralising the inhibitor in serum was raised in rabbits.

Separation of Heparin into Subtractions by DEAE-Cellulose Chromatography

1979 ◽  
Vol 42 (05) ◽  
pp. 1452-1459 ◽  
Author(s):  
Robert H Yue ◽  
Toby Starr ◽  
Menard M Gertler
Keyword(s):  
High Activity ◽  
Uronic Acid ◽  
Deae Cellulose ◽  
Exchange Chromatography ◽  
Net Charge ◽  
Uronic Acid Content ◽  
Successful Separation ◽  
Salt Gradient ◽  
Structure And Activity ◽  
Low Activity

SummaryCommercial porcine heparin can be separated into three distinct subtractions by using DEAE-cellulose chromatography and a stepped salt gradient. Gram quantities of heparin can be fractionated by this technique. All three heparin subtractions can accelerate the inhibition of thrombin by antithrombin III with different efficiency. The specific activities of the high activity heparin, intermediate activity heparin and low activity heparin are 228 units/mg, 142 units/mg and 95 units/mg, respectively. Both the uronic acid content and the quantity of N-SO4 for all three heparin subfractions have been evaluated. The high activity heparin has the lowest uronic acid and N-SO4 content. The successful separation of commercial heparin into three distinct subfractions by means of ion-exchange chromatography suggests that the net charge on these three heparin components will serve as a model system in the elucidation of the structure and activity relationship to the biological function of heparin.

scholarly journals Synthesis and Biological Evaluation of a Novel Glycidyl Metharcylate/Phaytic Acid-Based on Bagasse Xylan Composite Derivative

Polymers ◽  
2021 ◽  
Vol 13 (13) ◽  
pp. 2084
Author(s):  
Mingkun Li ◽  
Heping Li ◽  
Hongli Liu ◽  
Zhiming Zou ◽  
Chaoyu Xie
Keyword(s):  
Lung Cancer ◽  
Thermal Stability ◽  
Ionic Liquid ◽  
Molecular Docking ◽  
Phytic Acid ◽  
Biological Activities ◽  
Biological Evaluation ◽  
Potential Candidate ◽  
Reaction Conditions ◽  
Cell Proliferation Inhibition

The development of natural biomass materials with excellent properties is an attractive way to improve the application range of natural polysaccharides. Bagasse Xylan (BX) is a natural polysaccharide with various biological activities, such as antitumor, antioxidant, etc. Its physic-chemical and biological properties can be improved by functionalization. For this purpose, a novel glycidyl metharcylate/phytic acid based on a BX composite derivative was synthesized by a free radical polymerization technique with glycidyl metharcylate (GMA; GMABX) and further esterification with phytic acid (PA; GMABX-PA) in ionic liquid. The effects of the reaction conditions (i.e., temperature, time, initiator concentration, catalyst concentration, GMA concentration, PA concentration, mass of ionic liquid) on grafting rate(G), conversion rate(C) and degree of substitution(DS) are discussed. The structure of the composite material structure was confirmed by FTIR, 1H NMR and XRD. SEM confirmed the particle morphology of the composite derivative. The thermal stability of GMABX-PA was determined by TG-DTG. Molecular docking was further performed to study the combination mode of the GMABX-PA into the active site of two lung cancer proteins (5XNV, 2EB2) and a blood cancer protein (2M6N). In addition, tumor cell proliferation inhibition assays for BX, GMABX-PA were carried out using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetraz -olium bromide (MTT) method. The results showed that various reaction conditions exhibited favorable gradient curves, and that a maximum G of 56% for the graft copolymerization and a maximum DS of 0.267 can be achieved. The thermal stability was significantly improved, as demonstrated by the fact that there was still 60% residual at 800 °C. The molecular docking software generated satisfactory results with regard to the evaluated binding energy and combining sites. The inhibition ratio of GMABX-PA on NCI-H460 (lung cancer cells) reached 29.68% ± 4.45%, which is five times higher than that of BX. Therefore, the material was shown to be a potential candidate for biomedical applications as well as for use as a heat resistant material.

scholarly journals Chemical identity of tryptensin with angiotensin

1980 ◽  
Vol 187 (3) ◽  
pp. 647-653 ◽  
Author(s):  
K Arakawa ◽  
M Yuki ◽  
M Ikeda
Keyword(s):  
Amino Acid ◽  
Thin Layer ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Cation Exchange Chromatography ◽  
Human Plasma Protein ◽  
Plasma Protein Fraction ◽  
Layer Chromatography

Tryptensin, a vasopressor substance generated from human plasma protein fraction IV-4 by trypsin, has been isolated and the amino acid composition analysed. The procedures used for the isolation were: (a) adsorption of the formed tryptensin on Dowex 50W (X2; NH4+ form); (b) gel filtration through Sephadex G-25; (c) cation-exchange chromatography on CM-cellulose; (d) anion-exchange chromatography on DEAE-cellulose; (e) re-chromatography on CM-cellulose; (f) gel filtration on Bio-Gel P-2; (g) partition chromatography on high-pressure liquid chromatography. The homogeneity of the isolated tryptensin was confirmed by thin-layer chromatography and thin-layer electrophoresis. The amino acid analysis of the hydrolysate suggested the following proportional composition: Asp, 1; Val, 1; Ile, 1; Tyr, 1; Phe, 1; His, 1; Arg, 1; Pro, 1. This composition is identical with that of human angiotensin.

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