Notes: Separation of 13-and 9-Hydroperoxide Lyase Activities in Cotyledons of Cucumber Seedlings

1989 ◽  
Vol 44 (9-10) ◽  
pp. 883-885 ◽  
Author(s):  
Kenji Matsui ◽  
Yasushi Shibata ◽  
Tadahiko Kajiwara ◽  
Akikazu Hatanaka
Keyword(s):  
Ion Exchange ◽  
Specific Activity ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
The Other ◽  
Hydroperoxide Lyase ◽  
Cucumber Seedlings ◽  
Sh Reagents ◽  
Lyase Activity ◽  
Hydroperoxylinoleic Acid

Abstract In cucumber cotyledons, both C6- and C9- aldehyde were formed via hydroperoxide (HPO) lyase activity. Because it has not been elucidated whether these activities are attributed to one enzyme which can cleave both 13-and 9-HPO or to two or more enzymes each of which specifically cleaves 13-or 9-HPO , an attempt to separate HPO lyase activity was done. Ion exchange chromatography separated this activity into two fractions, one of which specifically cleaved 13-hydroperoxylinoleic acid and the other specifically cleaved the 9-isomer. 13-HPO-specific activity was most active at pH 8.0 and 9-HPO-specific one was at pH 6.5. SH -reagents inhibited both the lyases but to different extents.

Evaluation of dialysis treatment in uremic patients by gel filtration of serum

1990 ◽  
Vol 36 (11) ◽  
pp. 1906-1910 ◽  
Author(s):  
J Osada ◽  
T Gea ◽  
C Sanz ◽  
I Millan ◽  
J Botella
Keyword(s):  
Ion Exchange ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
The Other ◽  
Control Group ◽  
Dialysis Treatment ◽  
Additional Information ◽  
Molecular Masses ◽  
Two Peaks

Abstract A group of substances of molecular masses between 300 and 1500 Da have been found to be toxic metabolites in patients with uremia. We determined the concentration in serum of these molecules in the following groups of patients: two hemodialyzed groups (one with cuprophane and the other with polyacrylonitrile dialyzers), one group treated with continuous ambulatory peritoneal dialysis, one group of nondialyzed azotemic patients, and one control group of healthy persons. Ultrafiltrates of the subjects' sera were fractionated on Sephadex G-15 followed by ion-exchange chromatography. Eluates were monitored by absorbance at 254 and 206 nm. Partially characterized peaks P1 and P2, obtained by gel filtration, correlated with the concentration of creatinine in serum; their concentrations were significantly (P less than 0.01) larger in hemodialyzed groups than in peritoneal dialyzed or in nondialyzed azotemic patients. After ion-exchange chromatography, two peaks (P'5 and P'6) correlated with serum creatinine and also were larger in hemodialyzed patients than in the other groups. Apparently, adequate discrimination is obtained by gel-filtration analysis and further analysis by ion-exchange chromatography does not provide additional information in most of the affected patients.

scholarly journals Isolation, partial purification and characterization of phospholipase A2 from Naja Katiensis venom

2021 ◽  
Vol 13 (2) ◽  
pp. 107-112
Author(s):  
C.F. Okechukwu ◽  
P.L. Shamsudeen ◽  
R.K. Bala ◽  
B.G. Kurfi ◽  
A.M. Abdulazeez
Keyword(s):  
Ion Exchange ◽  
Phospholipase A2 ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Partial Purification ◽  
Crude Venom

The most effective and acceptable therapy for snakebite victims is the immediate administration of antivenin which is limited by problems of hypersensitivity reactions in some individuals and its inability to resolve the local effects of the venom. The aim of this study was to isolate, partially purify and characterize phospholipase A2 from Naja Katiensis venom. Phospholipase A2 was partially purified via a two-step process: gel filtration on Sephadex G-75 and ion exchange chromatography using CM Sephadex, and subjected to SDS-PAGE analysis. From the results, the specific activity of the partially purified PLA2 decreased from 0.67μmol/min/mg in crude venom to 0.29μmol/min/mg after ion exchange chromatography with a yield of 5% and purification fold of 0.43. The optimum temperature of the purified PLA2 was found to be 35ºC and optimum p.H of 7. velocity studies for the determination of kinetic constants using L-a-lecithin as substrate revealed a Km  of 1.47mg/ml and Vmax  of 3.32μ moles/min/mg. The sodium dodecyl sulphate polyacrylamide gel electrophoresis of the purified PLA2 showed a distinct band with molecular weight estimated to be 14KDa. In conclusion, the present study shows that phospholipase A2 was isolated, purified and characterized. This may serve as a promising candidate for future development of a novel anti-venin drug.

scholarly journals Isolation, in the intact state, of the pterin molybdenum cofactor from xanthine oxidase

1989 ◽  
Vol 263 (2) ◽  
pp. 477-483 ◽  
Author(s):  
J Deistung ◽  
R C Bray
Keyword(s):  
Aqueous Solution ◽  
Ion Exchange ◽  
Xanthine Oxidase ◽  
Specific Activity ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Molybdenum Cofactor ◽  
Anaerobic Conditions ◽  
Intact State

A procedure is described for isolation of the pterin molybdenum cofactor, in the active molybdenum-containing state, starting from purified milk xanthine oxidase. The method depends on the use of anaerobic-glove-cabinet techniques and on working in aqueous solution, in the presence of 1 mM-Na2S2O4. SDS was used to denature the protein, followed by ion-exchange chromatography and gel filtration. The cofactor, obtained at concentrations up to 0.5-1.0 mM, was fully active in the nit-1 assay [Hawkes & Bray (1984) Biochem. J. 214, 481-493], with a specific activity of 22 nmol of NO2-/min per pg-atom of Mo (with 15% molybdate-dependence). The Mr, determined by gel filtration, was about 610, consistent with the structure proposed by Kramer, Johnson, Ribeiro, Millington & Rajagopalan [(1987) J. Biol. Chem. 262, 16357-16363]. At pH 5.9, under anaerobic conditions, the cofactor was stable for at least 300 h at 20-25 degrees C.

scholarly journals Influence of various levels of L-asparaginase II purification on the cytotoxicity, DNA level, and apoptosis in Hep-2 cells

2010 ◽  
Vol 4 (2) ◽  
pp. 46-52
Author(s):  
Hayfa H. Hassani ◽  
Rawa'a J. Toma
Keyword(s):  
Ion Exchange ◽  
Cancer Cells ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
The Other ◽  
Proteus Vulgaris ◽  
Bacterial Enzyme ◽  
M Phase ◽  
Induced Apoptosis

The genetic effects of several concentrations of L–Asparaginase II (ASNase II), produced by Proteus vulgaris strain Pv.U92, at various levels of purification (ultrasonication, precipitation, ion-exchange chromatography and gel filtration chromatography) on cancer cells line of Hep–2 were studied. This bacterial enzyme with concentration 4 U/ml at gel filtration level was revealed a putative cytotoxicity against cancer cells in comparison with other concentrations and steps of purification were used in this work. Moreover, 4 U/ml of ASNase II at gel purification level has a distinguished role on arrest cancer cells division of Hep–2; it was reduced the content of DNA at each phase of cancer cell cycle particularly at G2/M phase, the level of DNA was 3%. On the other hand, the partial purified enzyme, L–ASNase II, was induced apoptosis by both levels of purification ion–exchange and gel filtration, the apoptotic fractionation was 0.86 and 0.7 respectively .

Isolation of a cationic peroxidase from cultured peanut cells

1980 ◽  
Vol 58 (21) ◽  
pp. 2280-2284 ◽  
Author(s):  
B. A. Maldonado ◽  
R. B. van Huystee
Keyword(s):  
Ion Exchange ◽  
Suspension Culture ◽  
Cell Suspension ◽  
Ammonium Sulfate ◽  
Cell Suspension Culture ◽  
Specific Activity ◽  
High Specific Activity ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Single Peptide

Medium that had supported the growth of a peanut cell suspension culture was found to be a rich source of peroxidase. Precipitation with acetone and ammonium sulfate, followed by a pH shift and ion exchange chromatography, led to the isolation of a single cationic peroxidase isozyme of high specific activity and high RZ of 2.91. The isolated peroxidase was shown to be a single peptide chain.

scholarly journals Purification and Partial Characterization of a Murein Hydrolase, Millericin B, Produced by Streptococcus milleri NMSCC 061

2000 ◽  
Vol 66 (1) ◽  
pp. 23-28 ◽  
Author(s):  
M. Beukes ◽  
G. Bierbaum ◽  
H.-G. Sahl ◽  
J. W. Hastings
Keyword(s):  
Ion Exchange ◽  
Specific Activity ◽  
Gel Filtration ◽  
Fold Increase ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Amino Acid Sequences ◽  
Antimicrobial Substances ◽  
Streptococcus Milleri ◽  
N Terminus

ABSTRACT Streptococcus milleri NMSCC 061 was screened for antimicrobial substances and shown to produce a bacteriolytic cell wall hydrolase, termed millericin B. The enzyme was purified to homogeneity by a four-step purification procedure that consisted of ammonium sulfate precipitation followed by gel filtration, ultrafiltration, and ion-exchange chromatography. The yield following ion-exchange chromatography was 6.4%, with a greater-than-2,000-fold increase in specific activity. The molecular weight of the enzyme was 28,924 as determined by electrospray mass spectrometry. The amino acid sequences of both the N terminus of the enzyme (NH2 SENDFSLAMVSN) and an internal fragment which was generated by cyanogen bromide cleavage (NH2 SIQTNAPWGL) were determined by automated Edman degradation. Millericin B displayed a broad spectrum of activity against gram-positive bacteria but was not active againstBacillus subtilis W23 or Escherichia coli ATCC 486 or against the producer strain itself. N-Dinitrophenyl derivatization and hydrazine hydrolysis of free amino and free carboxyl groups liberated from peptidoglycan digested with millericin B followed by thin-layer chromatography showed millericin B to be an endopeptidase with multiple activities. It cleaves the stem peptide at the N terminus of glutamic acid as well as the N terminus of the last residue in the interpeptide cross-link of susceptible strains.

Specific activity and isotope abundances of strontium in purified strontium-82

2016 ◽  
Vol 31 (2) ◽  
pp. 458-463 ◽  
Author(s):  
J. M. Fitzsimmons ◽  
D. G. Medvedev ◽  
L. F. Mausner
Keyword(s):  
Ion Exchange ◽  
Linear Accelerator ◽  
Specific Activity ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Rubidium Chloride

A linear accelerator was used to irradiate a rubidium chloride target with protons to produce strontium-82 (Sr-82), and the Sr-82 was purified by ion exchange chromatography.

scholarly journals Purification and Characterization of Cellulase from Termite Ametermes eveuncifer (Silverstri) Soldiers

2016 ◽  
Vol 9 (1) ◽  
pp. 1 ◽  
Author(s):  
B. S. Fagbohunka ◽  
R. E. Okonji ◽  
Ayinla Zainab Adenike
Keyword(s):  
Ion Exchange ◽  
Specific Activity ◽  
Ion Exchange Chromatography ◽  
Maximum Activity ◽  
Exchange Chromatography ◽  
Purification And Characterization ◽  
Ammonium Sulphate Precipitation ◽  
Percentage Yield ◽  
Termite Soldiers

Cellulase enzyme was purified and characterized from termite soldiers (Ametermes eveuncifer) using 70% ammonium sulphate precipitation, ion exchange chromatography and affinity chromatography. The enzyme isolated had a specific activity of 5.04 U/mg with a percentage yield of 11.7%. The enzyme showed maximum activity at 500C and pH 8. The enzyme was not inhibited by Ba2+ at a concentration of 1mM and Pb2+ at 10 mM concentration but was inhibited by other metal ions at 1 mM and 10 mM concentrations of their salts (NaCl, KCl, MnCl2, and NiCl2),

scholarly journals Separation of rat lactate dehydrogenase isoenzyme C4 from other isoenzymes by affinity and ion-exchange chromatography

1981 ◽  
Vol 199 (1) ◽  
pp. 75-79 ◽  
Author(s):  
A A Ansari
Keyword(s):  
Lactate Dehydrogenase ◽  
Ion Exchange ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
The Other ◽  
Molecular Forms ◽  
Dehydrogenase Isoenzyme ◽  
Lactate Dehydrogenase C ◽  
B Subunits

Lactate dehydrogenase C, an isoenzyme composed of C polypeptide subunits and found only in mature testes and spermatozoa, differs kinetically, chemically and immunologically from the five common isoenzymes of lactate dehydrogenase, each of which is a tetramer of A and/or B subunits. In the rat lactate dehydrogenase C exists in two molecular forms, isoenzymes C4 and A1C3. In addition to these two forms of lactate dehydrogenase C, rat testicular homogenate contains all the five isoenzymes of A and B type. Purification of isoenzyme C4 requires its separation from the other six isoenzymes, of which isoenzymes A1C3 and A3B1 are the most difficult ones to separate. In the present study isoenzyme A3B1, along with other enzymes, was separated from isoenzyme C4 by AMP-Sepharose chromatography by using a gradient of increasing concentration of NAD+-pyruvate adduct. In the next step, isoenzyme A1C3 was separated from isoenzyme C4 by DEAD-cellulose chromatography, resulting in a pure lactate dehydrogenase isoenzyme C4 preparation.

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