Isolation of Prolidase from Amniotic Fluid and Study of its Kinetic and Affinity Properties Towards Pharmacological Compounds
<span>The research included the isolation of prolidase from human amniotic fluid using different biochemical techniques, One proteinous peak had been isolated by gel filtration using sephadex )G-50) and from sephadex (G-100) that produced by ammonium sulphate precipitation (60%) after dialysis...
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College of Education for Pure Sciences,
2019-09-01T00:00:00Z.
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| Summary: | <span>The research included the isolation of prolidase from human amniotic fluid using different biochemical techniques, One proteinous peak had been isolated by gel filtration using sephadex )G-50) and from sephadex (G-100) that produced by ammonium sulphate precipitation (60%) after dialysis. The approximately molecular weight of the enzyme using gel filtration chromatography (G-100) was (52269.7) Dalton and specific activity of 13516.6 unit/mg protein with 75 fold of purification. The results showed that the optimum conditions of purified enzyme from amniotic fluid were at (50 </span><span>µ</span><span>g/ml) of protein as a source of the enzyme using (60 mM/L) Tris-HCl buffer solution at pH (8.0) act for (32) minutes at (39°C). Using Line Weaver-Burk plot, the values of maximum velocity (</span><span>V</span><span><span>max</span></span><span>) and Michaelis constant (</span><span>K</span><span><span>m</span></span><span>) were found to be (3448.2 U/L) and (10 mM/L) respectively using Gly-Pro as a substrate. </span><br /> <span>Finally, also, involved the study of the effect pharmacological chemicals compounds on the enzyme activity, the results showed that the drug chemical compounds for type ceramide, metoclopramide, pseudoephedrine, diphenhydramine-HCl, chloramphnicol, paracetamol and allopurinol give the inhibition type competitive, with increased in </span><span>K</span><span><span>m</span></span><span> value to 66.6, 71.42, 52.63, 55.55, 83.33, 90.9 and100.0 mM/L respectively for inhibitors above, while the others pharmacological compounds for theophlline anhydrous, caffeine anhydrous, metronidazole and chlorpheniramine maleate, give the inhibition type of non-competitive with decreased in </span><span>V</span><span><span>max</span></span><span> values to 2173.9, 2439.0, 2000.0 and 2325.58 U/L respectively, but dexathamazone was an activator to the prolidase approximately 27.18 U/L.</span> |
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| Item Description: | 1812-125X 2664-2530 10.33899/edusj.2019.162958 |