Rapid measurement of leucine-specific activity in biological fluids by ion-exchange chromatography and post-column ninhydrin detection

1991 ◽  
Vol 571 (1-2) ◽  
pp. 29-36 ◽  
Author(s):  
E.Patrick Donahue ◽  
Laurel L. Brown ◽  
Paul J. Flakoll ◽  
Naji N. Abumrad
Keyword(s):  
Ion Exchange ◽  
Specific Activity ◽  
Biological Fluids ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Rapid Measurement

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.

Automated analysis for taurine in biological fluids and tissues.

1987 ◽  
Vol 33 (6) ◽  
pp. 835-837 ◽  
Author(s):  
H O Goodman ◽  
Z K Shihabi
Keyword(s):  
Ion Exchange ◽  
Continuous Flow ◽  
Biological Fluids ◽  
Automated Analysis ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Fluorescent Compound ◽  
Automated Method ◽  
Tissue Homogenates ◽  
Proteins And Peptides

Abstract We have developed an automated method of analysis for taurine, based on incorporating an ion-exchange chromatography column into the continuous-flow AutoAnalyzer (Technicon). After removal of proteins and peptides by dialysis, taurine is selectively eluted from an ion-exchange column and reacted with o-phthaldialdehyde to yield a fluorescent compound. The advantages of this method are: full automation with no need for sample deproteinization or cleanup; sensitivity, detecting as little as 5 mumol/L; speed (20 samples per hour); and flexibility. It can be used for assaying taurine in urine, plasma, cerebrospinal fluid, and tissue homogenates. This method can be adapted for assays of other metabolites.

Effect of Deproteinating Agents (Picric Acid and Sulfosalicylic Acid) on Analysis for Free Amino Acids in Swine Blood and Tissue

1969 ◽  
Vol 52 (5) ◽  
pp. 981-984 ◽  
Author(s):  
J E Knipfel ◽  
D A Christensen ◽  
B D Owen
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Ion Exchange ◽  
Free Amino ◽  
Biological Fluids ◽  
Picric Acid ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Sulfosalicylic Acid ◽  
Coefficients Of Variation

Abstract Amino acid analyses were performed on samples of blood, liver tissue, loin muscle, and ham muscle by ion exchange chromatography after deproteination of the samples with picric acid or sulfosalicylic acid (SSA). Resolution of threonine and serine from the ion exchange column was poor when SSA was used as the deproteinating agent. Twelve of sixteen amino acids were higher (P < 0.05) in serum deproteinated with picric acid as compared to concentrations determined after SSA deproteination. Amino acid values for ham muscle tended to be higher after deproteination with picric acid; however, with liver and loin muscle samples, the values were somewhat higher after SSA deproteination. In both serum and tissue analyses, coefficients of variation were lower for niGSt amino acids when picric acid was utilized as the deproteinating agent. The latter observation, in particular, suggests that picric acid is preferable to SSA as a deproteinating agent before amino acid analyses of biological fluids. Standardization of methods of deproteination is needed to allow meaningful comparisons of data.

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.

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.

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),

Sign in / Sign up

Export Citation Format

Share Document