Folic acid conjugase from Plasma. I. Partial purification and properties.
Human plasma conjugase was partially purified by ammonium sulphate and DEAE-cellulose column fractionation. The pH optimum was found to be 4.5. The enzyme followed normal Michaelis-Menten Kinetics. The Km value was found to be 1.4 x 10(-7)M for folylheptaglutatmate assuming this compound to be the predominant form of folate in yeast extract. The enzyme required sulfhydryl compounds for full activity. Differential microbiological assay of the conjugase reaction product using L. casei and S. faecalis as the test organisms revealed that during the initial stage of the reaction considerable portion of the reaction product was at polyglutamyl stage. On prolonged incubation polyglutamates decreased due to further hydrolysis of the gamma-glutamyl residues. The plasma conjugase activity of different species of animals showed wide variations. Goat, dog and rabbit plasma had neglibible activity while monkey, guinea pig and chick plasma had moderate activity. Bovine plasma had low activity while rat plasma had high activity. The plasma conjugase activity of normal Indian adults ranged from 400 to 900 units per ml, and there was no significant sex difference. The values obtained are considerably higher than those reported in literature.
D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology; D001769:Blood; D002268:Carboxypeptidases / Q000302:isolation & purification*; D002417:Cattle; D002645:Chickens; D003545:Cysteine / Q000494:pharmacology; D004285:Dogs; D005260:Female; D005492:Folic Acid / Q000097:blood / Q000302:isolation & purification*; D005978:Glutathione / Q000494:pharmacology; D006041:Goats; D006168:Guinea Pigs; D000882:Haplorhini; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008297:Male; D008623:Mercaptoethanol / Q000494:pharmacology; D011817:Rabbits; D051381:Rats; D013045:Species Specificity; D013268:Stimulation, Chemical; D011623:gamma-Glutamyl Hydrolase / Q000097:blood / Q000302:isolation & purification*
D008623:Mercaptoethanol; D005492:Folic Acid; D002268:Carboxypeptidases; D011623:gamma-Glutamyl Hydrolase; D005978:Glutathione; D003545:Cysteine; D001205:Ascorbic Acid