Purification and characterization of catalase from sprouted black gram (Vigna mungo) seeds.
TL;DR: The enzyme showed optimal activity at a temperature of 40°C and pH 7.0 and had a half life of 7h 30 min at 50°C, and the effect of inhibitors like Sodium azide and EDTA and different metal ions on catalase activity were studied.
About: This article is published in Journal of Chromatography B.The article was published on 2012-03-15. It has received 25 citations till now. The article focuses on the topics: Catalase & Vigna.
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TL;DR: The direct and indirect involvement of deficiency and/or modification of catalase in the pathogenesis of some important diseases such as diabetes mellitus, Alzheimer's disease, Parkinson’s disease, vitiligo, and acatalasemia is described.
Abstract: Reactive species produced in the cell during normal cellular metabolism can chemically react with cellular biomolecules such as nucleic acids, proteins, and lipids, thereby causing their oxidative modifications leading to alterations in their compositions and potential damage to their cellular activities. Fortunately, cells have evolved several antioxidant defense mechanisms (as metabolites, vitamins, and enzymes) to neutralize or mitigate the harmful effect of reactive species and/or their byproducts. Any perturbation in the balance in the level of antioxidants and the reactive species results in a physiological condition called "oxidative stress." A catalase is one of the crucial antioxidant enzymes that mitigates oxidative stress to a considerable extent by destroying cellular hydrogen peroxide to produce water and oxygen. Deficiency or malfunction of catalase is postulated to be related to the pathogenesis of many age-associated degenerative diseases like diabetes mellitus, hypertension, anemia, vitiligo, Alzheimer's disease, Parkinson's disease, bipolar disorder, cancer, and schizophrenia. Therefore, efforts are being undertaken in many laboratories to explore its use as a potential drug for the treatment of such diseases. This paper describes the direct and indirect involvement of deficiency and/or modification of catalase in the pathogenesis of some important diseases such as diabetes mellitus, Alzheimer's disease, Parkinson's disease, vitiligo, and acatalasemia. Details on the efforts exploring the potential treatment of these diseases using a catalase as a protein therapeutic agent have also been described.
372 citations
TL;DR: This review addresses the developments, limitations, and considerations of IMAC in the biopharmaceutical industry.
Abstract: After 35 years of development, immobilized metal ion affinity chromatography (IMAC) has evolved into a popular protein purification technique. This review starts with a discussion of its mechanism and advantages. It continues with its applications which include the purification of histidine-tagged proteins, natural metal-binding proteins, and antibodies. IMAC used in conjunction with mass spectroscopy for phosphoprotein fractionation and proteomics is also covered. Finally, this review addresses the developments, limitations, and considerations of IMAC in the biopharmaceutical industry.
200 citations
Cites background from "Purification and characterization o..."
...Catalase from Vigna mungo seeds (Kandukuri et al. 2012) Antibody Anti-TNP IgG mouse MAbs (Bresolin et al. 2010)....
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TL;DR: In this article, an interdisciplinary review covers the updated information about the natural and the synthetic materials based on polymers or low molecular compounds, organic or inorganic ones, utilized by researchers to immobilize the most popular enzyme named CATALASE.
112 citations
TL;DR: The results demonstrate that this edible plant, Rumex obtusifolius L., might be a potential source of natural antioxidants with good antioxidant enzyme capacity.
Abstract: Aims: The purpose of this study was to measure antioxidant enzyme (polyphenol oxidase, peroxidase, catalase and superoxide dismutase) activities of crude extract of Rumex obtusifolius L. in order to gain insight about this plant’s antioxidant potential. Study Design: The study was composed of the collection of plant material, extractions of the antioxidant enzymes, activity measurements of the enzymes and finally evaluation of the experimental results. Place and Duration of Study: Department of Chemistry (biochemistry laboratories), Faculty of Science and Arts of Sakarya University, between June 2015 and July 2015. Methodology: Enzymatic antioxidant activity of this plant was investigated by carrying out catalase, superoxide dismutase, peroxidase and polyphenol oxidase enzyme activity assays. Enzyme activities of the crude extract were measured by using spectrophotometric method. Optimum pH and temperature values of each enzyme were also determined for measurement of enzyme activities in ideal conditions. Results: Finally, our results showed that Rumex obtusifolius L. crude extract had good activity for all the enzymatic procedures tested. The activity levels of enzymatic antioxidants polyphenol oxidase, peroxidase, catalase and superoxide dismutase of the plant were found to be 12.8; 195.2; Original Research Article Alici and Arabaci; ARRB, 11(3): 1-7, 2016; Article no.ARRB.29809 2 38.7; 11.6 EU/mL, respectively. Optimum pH and temperature values of all the enzymes (except PPO: optimum temperature 30°C) tested were also foun d to be 7.0 and 25°C, respectively. Conclusion: Our results demonstrate that this edible plant, Rumex obtusifolius L., might be a potential source of natural antioxidants with good antioxidant enzyme capacity.
78 citations
Additional excerpts
...8 30 35 20 Black gram (Vigna mungo) [21] CAT Hydrogen peroxide 7....
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TL;DR: TPP is a simple, economical, and quick method for the recovering of catalase and can be used for the purification process, and is shown to be effective in potato crude extract purification.
Abstract: Three-phase partitioning (TPP) was used to purify and recover catalase from potato crude extract. The method consists of ammonium sulfate saturation, t-butanol addition, and adjustment of pH, respectively. The best catalase recovery (262 %) and 14.1-fold purification were seen in the interfacial phase in the presence of 40 % (w/v) ammonium sulfate saturation with 1.0:1.0 crude extract/t-butanol ratio (v/v) at pH 7 in a single step. The sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the enzyme showed comparatively purification and protein molecular weight was nearly found to be 56 kDa. This study shows that TPP is a simple, economical, and quick method for the recovering of catalase and can be used for the purification process.
41 citations
References
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TL;DR: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products.
Abstract: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products. Four major components of the head are cleaved during the process of assembly, apparently after the precursor proteins have assembled into some large intermediate structure.
232,912 citations
TL;DR: This assay is very reproducible and rapid with the dye binding process virtually complete in approximately 2 min with good color stability for 1 hr with little or no interference from cations such as sodium or potassium nor from carbohydrates such as sucrose.
225,085 citations
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TL;DR: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products as mentioned in this paper.
Abstract: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products. Four major components of the head are cleaved during the process of assembly, apparently after the precursor proteins have assembled into some large intermediate structure.
203,017 citations
TL;DR: It seems possible that one factor in aging may be related to deleterious side attacks of free radicals (which are normally produced in the course of cellular metabolism) on cell constituents.
Abstract: The phenomenon of growth, decline and death—aging—has been the source of considerable speculation (1, 8, 10). This cycle seems to be a more or less direct function of the metabolic rate and this in turn depends on the species (animal or plant) on which are superimposed the factors of heredity and the effects of the stresses and strains of life—which alter the metabolic activity. The universality of this phenomenon suggests that the reactions which cause it are basically the same in all living things. Viewing this process in the light of present day free radical and radiation chemistry and of radiobiology, it seems possible that one factor in aging may be related to deleterious side attacks of free radicals (which are normally produced in the course of cellular metabolism) on cell constituents.* Irradiation of living things induces mutation, cancer, and aging (9). Inasmuch as these also arise spontaneously in nature, it is natural to inquire if the processes might not be similar. It is believed that one mechanism of irradiation effect is through liberation of OH and HO 2 radicals (12). There is evidence, although indirect, that these two highly active free radicals are produced normally in living systems. In the first place, free radicals are present in living cells; this was recently demonstrated in vivo by a paramagnetic resonance absorption method (3). Further, it was shown that the concentration of free radicals increased with increasing metabolic activity in conformity with the postulates set forth some years ago that free radicals were involved in biologic oxidation-reduction reactions (11, 13). Are some of these free radicals OH and/or HO2, or radicals of a similar high order of reactivity, and where might they arise in the cell? The most likely source of OH and HO2 radicals, at least in the animal cell, would be the interaction of the respiratory enzymes involved
7,917 citations
TL;DR: A quantitative, spectrophotometric technique for following the breakdown of hydrogen peroxide has been developed for routine studies of catalase kinetics and appears to give lower values forCatalase activity than do titration techniques.
6,007 citations