scholarly journals Partial Purification and Characterization of Catalase from Banana Peels

2016 ◽  
Vol 13 (2) ◽  
pp. 392-398
Author(s):  
Baghdad Science Journal
Keyword(s):  
Gel Filtration ◽  
Maximum Velocity ◽  
Kinetic Studies ◽  
Maximal Activity ◽  
Partial Purification ◽  
Purification And Characterization ◽  
Km Value ◽  
Almost All ◽  
Kinetics Of

Catalase (EC 1.11.1.6) is a well known enzyme which exists in almost all living creatures exposing to oxygen (such as plants, bacteria, and animals). It is a very necessary enzyme to protect the cell from oxidative detriment by reactive oxygen species (ROS). The aim of this study is the partial purification and characterization of Catalase enzyme from Banana peels. In this study, fresh banana peels are treated with 70 % ethanol ,further separated with chloroform ,water and ethyl acetate respectively .The supernatant of the enzymatic sample which is treated with chloroform is loaded into gel filtration column with Sephadex G-100 (1.0 x 90 cm) equilibrated with pH7 buffer media (phosphate buffer 0.1 M). Kinetic studies of the purified enzyme activity are measured and characterized .The maximal activity (26.04 units/mg) of catalase is observed with chloroform buffer extraction. The kinetics of catalase; Michalis constant Km and maximum velocity Vmax is determined using Linweaver- Burk plot, The Km value for catalase (434.7mM), Vmax (100 m mole min -1). Characterization results demonstrate that the optimal pH for activity is (7.6). And the optimal temperature for activity is 30?C .The present study indicates that Banana peels is a good source of catalase enzyme.

scholarly journals Purification and Characterization of Beta-Amylase of Bacillus subtilis Isolated from Kolanut Weevil

2012 ◽  
Vol 4 (1) ◽  
Author(s):  
Femi-Ola Titilayo Olufunke ◽  
Ibikunle Ibidapo Azeez
Keyword(s):  
Bacillus Subtilis ◽  
Acid Treatment ◽  
Gel Filtration ◽  
Maximum Velocity ◽  
Maximal Activity ◽  
Purification And Characterization ◽  
Michaelis Constant ◽  
Optimal Activity ◽  
Ethylene Diaminetetraacetic Acid

An extracellular beta amylase was induced in cultures of Bacillus subtilis isolated from the kolanut weevil, Balanogastris kolae grown in liquid medium that contained kolanut starch as sole carbon source. The enzyme was partially purified 1.28-fold by acid treatment with ice cold 1.0N HCl and 6.4-fold by gel filtration with Sephadex G-150. The beta amylase had a molecular weight of 39.4 kDa .The enzyme had its optimal activity at pH 5.0 and exhibited maximal activity at temperature of 50oC. The activity of the enzyme was enhanced by Na+, Ca2+  and ethylene diaminetetraacetic acid (EDTA), while Hg2+ ,Mg2+ and  Fe2+ acted as inhibitors of its activity. The beta amylase had an apparent Michaelis constant Km of 5.0 mg/ml and maximum velocity (Vmax) of 50 U/mg proteins.

scholarly journals Partial purification and characterization of an endo-α-N-acetylgalactosaminidase from the culture medium of Streptomyces sp. OH-11242

1992 ◽  
Vol 288 (2) ◽  
pp. 475-482 ◽  
Author(s):  
I Ishii-Karakasa ◽  
H Iwase ◽  
K Hotta ◽  
Y Tanaka ◽  
S Omura
Keyword(s):  
Enzyme Activity ◽  
Culture Medium ◽  
Gel Filtration ◽  
Partial Purification ◽  
Gastric Mucus ◽  
Purification And Characterization ◽  
Streptomyces Sp ◽  
Optimum Ph ◽  
New Type

For the purification of a new type of endo-alpha-N-acetylgalactosaminidase from the culture medium of Streptomyces sp. OH-11242 (endo-GalNAc-ase-S) [Iwase, Ishii, Ishihara, Tanaka, Omura & Hotta (1988) Biochem. Biophys. Res. Commun. 151, 422-428], a method for assaying enzyme activity was established. Using purified pig gastric mucus glycoprotein (PGM) as the substrate, oligosaccharides liberated from PGM were pyridylaminated, and the reducing terminal sugars of oligosaccharides larger than Gal beta 1-3GalNAc were analysed by h.p.1.c. The crude enzyme of endo-GalNAc-ase-S was prepared as an 80% (w/v) ammonium sulphate precipitate from the concentrated culture medium. The enzyme was partially purified by gel chromatofocusing and subsequent DEAE-Toyopearl chromatography. Endo-enzyme activity eluted around pI 4.8 on a gel chromatofocusing column and eluted with 0.19-0.25 M-NaCl on a DEAE-Toyopearl column. In the enzyme fraction obtained, no exo-glycosidases or proteases could be detected. The molecular mass of the enzyme was estimated as 105 kDa by gel filtration, and the optimum pH was 5.5. Endo-GalNAc-ase-S hydrolysed the O-glycosidic linkage between GalNAc and Ser (Thr) in 3H-labelled and unlabelled asialofetuin, liberating both the disaccharide (Gal beta 1-3GalNAc) and the tetrasaccharide [Gal beta 1-3 (Gal beta 1-4GlcNAc beta 1-6)GalNAc]. When endo-alpha-N-acetylgalactosaminidase from Alcaligenes sp. (endo-GalNac-ase-A) was incubated with 3H-labelled and unlabelled asialofetuin, only the disaccharide (Gal beta 1-3GalNAc) was liberated.

scholarly journals Partial purification and characterization of xylanase from Bacillus cereus X3

2014 ◽  
Vol 11 (2) ◽  
pp. 1056-1061
Author(s):  
Baghdad Science Journal
Keyword(s):  
Bacillus Cereus ◽  
Maximal Activity ◽  
Partial Purification ◽  
Purification And Characterization ◽  
Xylanase Production ◽  
Optimum Activity ◽  
Biology Department ◽  
Characterization Study ◽  
Quantitative Screening

Three strain of Bacillus cereus were obtained from soil sours Laboratories of Biology Department/ College of Science/ University of Baghdad. The bacteria secreted extracellular xylanase in liquid cultur the test ability of xylanase production from these isolates was studied semi quantitative and quantitative screening appeared that Bacillus cereus X3 was the highest xylanase producer. The enzyme was partial purification 191 fold from cultur by reached step by 4 U/mg proteins by ammonium sulfat precipitation 80%, Ion exchang DEAE-cellulos chromatography Characterization study of the partial purifation enzyme revealed that the enzyme had a optimum activity pH8 and activity was stable in the pH rang (8-10) for 30min. maximal activity was attained at 50C

scholarly journals Optimization of Growth Conditions for Purification and Production of L-Asparaginase bySpirulina maxima

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Hanaa H. Abd El Baky ◽  
Gamal S. El Baroty
Keyword(s):  
Enzyme Activity ◽  
Gel Filtration ◽  
Growth Conditions ◽  
Partial Purification ◽  
Specific Enzyme ◽  
Purification And Characterization ◽  
Spirulina Maxima ◽  
Optimum Ph ◽  
Biomass Yields

L-asparaginase (L-AsnA) is widely distributed among microorganisms and has important applications in medicine and in food technology sectors. Therefore, the ability of the production, purification, and characterization of AsnA fromSpirulina maxima(SM) were tested. SM cultures grown in Zarrouk medium containing different N2(in NaNO3form) concentrations (1.25, 2.50, and 5.0 g/L) for 18 days contained a significant various quantity of dry biomass yields and AsnA enzyme levels. MS L-AsnA activity was found to be directly proportional to the N2concentration. The cultures of SM at large scales (300 L medium, 5 g/L N2) showed a high AsnA enzyme activity (898 IU), total protein (405 mg/g), specific enzyme activity (2.21 IU/mg protein), and enzyme yield (51.28 IU/L) compared with those in low N2cultures. The partial purification of crude MS AsnA enzyme achieved by 80% ammonium sulfate AS precipitated and CM-Sephadex C-200 gel filtration led to increases in the purification of enzyme with 5.28 and 10.91 times as great as that in SM crude enzymes. Optimum pH and temperature of purified AsnA for the hydrolyzate were 8.5 and 37 ± 0.2°C, respectively. To the best of our knowledge, this is the first report on L-asparaginase production inS. maxima.

scholarly journals Partial purification and characterization of chick-embryo prolyl 3-hydroxylase

1979 ◽  
Vol 183 (2) ◽  
pp. 303-307 ◽  
Author(s):  
K Tryggvason ◽  
K Majamaa ◽  
J Risteli ◽  
K I Kivirikko
Keyword(s):  
Molecular Weight ◽  
Chick Embryo ◽  
Ethylene Glycol ◽  
Affinity Chromatography ◽  
Concanavalin A ◽  
Gel Filtration ◽  
Partial Purification ◽  
Purification And Characterization ◽  
Chick Embryo Extract

Prolyl 3-hydroxylase was purified up to about 5000-fold from an (NH4)2SO4 fraction of chick-embryo extract by a procedure consisting of affinity chromatography on denatured collagen linked to agarose, elution with ethylene glycol and gel filtration. The molecular weight of the purified enzyme is about 160000 by gel filtration The enzyme is probably a glycoprotein, since (a) its activity is inhibited by concanavalin A, and (b) the enzyme is bound to columns of this lectin coupled to agarose and can be eluted with a buffer containing methyl alpha-D-mannoside. The Km values for Fe2+, 2-oxoglutarate, O2 and ascorbate in the prolyl 3-hydroxylase reaction were found to be very similar to those previously reported for these co-substrates in the prolyl 4-hydroxylase and lysyl hydroxylase reactions.

scholarly journals Partial Purification and Characterization of Inulinase Produced from a local isolate of Aspergillus niger J3

2010 ◽  
Vol 4 (2) ◽  
pp. 78-85
Author(s):  
Jasim M. Awdaa
Keyword(s):  
Aspergillus Niger ◽  
Gel Filtration ◽  
Partial Purification ◽  
Mercury Chloride ◽  
Purification And Characterization ◽  
Original Activity ◽  
Local Isolate ◽  
Optimum Ph ◽  
Final Purification

Inulinase was produced from local isolate of Aspergillus niger J3. The inulinase was purified by two steps included precipitation by amonium sulphate at (30-80) % sa-turation and gel filtration on sephadex G100. The final purification folds and the yield of the enzyme were 3.15 times and 28.24%, respectively. The purified enzyme has the following characteristics: The optimum pH of the enzyme activity was 5.5. The enzyme was most stable at pH (4.5 - 6). The optimum temperature for its activity was 45c. The enzyme retained its original activity when incubates at (30-55) c for 20 minutes. Mercury chloride inhibited the enzyme completely at concentration of 10mM, cupper sulphate and calisium chloride inhibited the enzyme at concentrations of 85% and 7% respectively. It was revealed that the enzyme had the efficiency to hydrolyze 87% of 5% inulin solution when treated at 45c for 120 min.

scholarly journals Innate Immunity in Lobsters: Partial Purification and Characterization of a Panulirus cygnus Anti-A Lectin

2012 ◽  
Vol 2012 ◽  
pp. 1-5 ◽  
Author(s):  
Robert L. P. Flower
Keyword(s):  
Innate Immunity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Spiny Lobster ◽  
Gel Filtration ◽  
Partial Purification ◽  
Purification And Characterization ◽  
Rat Erythrocytes ◽  
Group A ◽  
Panulirus Cygnus

A lectin detected in haemolymph from the Australian spiny lobster Panulirus cygnus agglutinated human ABO Group A cells to a higher titre than Group O or B. The lectin also agglutinated rat and sheep erythrocytes, with reactivity with rat erythrocytes strongly enhanced by treatment with the proteolytic enzyme papain, an observation consistent with reactivity via a glycolipid. The lectin, purified by affinity chromatography on fixed rat-erythrocyte stroma, was inhibited equally by N-acetylglucosamine and N-acetylgalactosamine. Comparison of data from gel filtration of haemolymph (behaving as a 1,800,000 Da macromolecule), and polyacrylamide gel electrophoresis of purified lectin (a single 67,000 Da band), suggested that in haemolymph the lecin was a multimer. The purified anti-A lectin autoprecipitated unless the storage solution contained chaotropic inhibitors (125 mmol/L sucrose: 500 mmol/L urea). The properties of this anti-A lectin and other similar lectins are consistent with a role in innate immunity in these invertebrates.

scholarly journals Purification and characterization of N-acetylglutamate 5-phosphotransferase from pea (Pisum sativum) cotyledons

1981 ◽  
Vol 195 (1) ◽  
pp. 71-81 ◽  
Author(s):  
G McKay ◽  
P D Shargool
Keyword(s):  
Pisum Sativum ◽  
Polyacrylamide Gel Electrophoresis ◽  
Deae Cellulose ◽  
Kinetic Studies ◽  
Exchange Chromatography ◽  
Purification And Characterization ◽  
Coomassie Blue ◽  
Km Value ◽  
Acetylglutamate Kinase

N-Acetylglutamate 5-phosphotransferase (acetylglutamate kinase, EC 2.7.2.8) has been isolated from pea (Pisum sativum) cotyledons and purified 312-fold by using heat treatment, (NH4)2SO4 fractionation, affinity chromatography on ATP--Sepharose and ion-exchange chromatography on DEAE-cellulose. This preparation was shown on polyacrylamide-gel electrophoresis to yield one band staining with Coomassie Blue. The enzyme was shown by a variety of techniques to be composed of two different kinds of subunits, of mol.wts. 43000 and 53000 respectively. These subunits are arranged to give either a dimeric or tetrameric enzyme composed of equal numbers of each type of subunit. The dimeric and tetrameric enzyme forms are thought to be interconvertible, the equilibrium between these forms being influenced by the type of ligand bound to the subunits. Kinetic studies performed on the purified enzyme, indicated a random Bi Bi type of mechanism. The enzyme displayed apparent negative co-operativity with respect to one of its substrates, N-acetylglutamate; as a result, two Km values were found for this substrate, one at 1.9 X 10(-3) M and the other at 6.2 X 10(-3) M. A single Km value for ATP was found to be 1.7 X 10(-3) M. Allosteric regulation by arginine was also shown. A model, based on the Koshland, Némethy & Filmer [(1966) Biochemistry 5, 365-385] Sequential model, which adequately describes the kinetic and structural properties of N-acetylglutamate 5-phosphotransferase, is presented.

Comparison of mitochondrial and cytosolic tRNA nucleotidyltransferases from Triticum aestivum

1999 ◽  
Vol 77 (2) ◽  
pp. 230-239
Author(s):  
Raffaela Vicaretti ◽  
Paul BM Joyce
Keyword(s):  
Triticum Aestivum ◽  
High Activity ◽  
Maximal Activity ◽  
Partial Purification ◽  
Elution Profile ◽  
Mitochondrial Enzyme ◽  
Alkaline Ph ◽  
Purification And Characterization ◽  
Purification Scheme

Here we report the partial purification and characterization of wheat mitochondrial ATP (CTP):tRNA nucleotidyltransferase (EC 2.7.7.25). Our purification scheme involves ammonium sulfate fractionation and chromatography on anion-exchange, hydroxyapatite, and affinity columns. Our results indicate that the enzyme is stable over a broad range of temperatures with highest activity at 37°C. High activity is seen at alkaline pH with a maximum at pH 9. The enzyme exhibits maximal activity in the presence of 10 mM MgCl2 and is inhibited by (at least) 100 mM NaCl. We also show that a second form of this enzyme exists in the wheat cytosolic fraction. This enzyme shares many features with the mitochondrial enzyme but differs from the mitochondrial enzyme in its elution profile from hydroxyapatite and in its response to manganese.Key words: tRNA nucleotidyltransferase, wheat, mitochondria.

scholarly journals The purification and characterization of glucokinase from the thermophile Bacillus stearothermophilus

1986 ◽  
Vol 237 (2) ◽  
pp. 415-420 ◽  
Author(s):  
C R Goward ◽  
R Hartwell ◽  
T Atkinson ◽  
M D Scawen
Keyword(s):  
Large Scale ◽  
Bacillus Stearothermophilus ◽  
Specific Activity ◽  
Gel Filtration ◽  
Exchange Chromatography ◽  
Purification And Characterization ◽  
Km Value ◽  
Extraneous Material ◽  
Sepharose 4B

Homogeneous glucokinase (EC 2.7.1.2) from the thermophile Bacillus stearothermophilus was isolated on the large scale by using four major steps: precipitation of extraneous material at pH 5.5, ion-exchange chromatography on DEAE-Sepharose, pseudo-affinity chromatography on Procion Brown H-3R-Sepharose 4B and gel filtration on Ultrogel AcA 34. The purified enzyme had a specific activity of about 330 units/mg of protein and was shown to exist as a dimer of subunit Mr 33,000. Kinetic parameters for the enzyme were determined with a variety of substrates. The glucokinase was highly specific for alpha-D-glucose, and the only other sugar substrate utilized was N-acetyl-alpha-D-glucosamine. The enzyme shows Michaelis-Menten kinetics, with a Km value of 150 microM for alpha-D-glucose. The glucokinase was maximally active at pH 9.0.

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