Arylesterase in serum: elaboration and clinical application of a fixed-incubation method.
A sensitive, specific, and simple method for determining serum or urine arylesterase (EC 3.1.1.2) is described. The enzyme acts on phenyl acetate to release phenol, which produces a stable indophenol dye with 4-aminoantipyrine and potassium ferricyanide. Arylesterase, a thiol enzyme, is reactivated by 2-mercaptoethanol and by cysteine, but not by reduced glutathione. Calcium is indispensable to stabilize and to activate (Km = 0.85 mmol/L) the enzyme; complete protection is achieved at CaCl2 20 mmol/L. Magnesium acts as a weak (Ki = 116 mmol/L), lanthanum as a potent (Ki = 5 mumol/L) competitive inhibitor. The activity is measured in diluted sera at phenyl acetate 4.0 mmol/L (Km = 1.12 mmol/L), pH 7.8 and 25 degrees C. The normal range extends from 53 to 186 kU/L, and four isoenzymes are present in sera from healthy adults. Arylesterase decreases in hepatic disorders, especially in cirrhosis and carcinoma of the liver, with reduction of the penultimate fraction in polyacryalmide gel electrophoresis.
D000293:Adolescent; D000328:Adult; D000367:Age Factors; D000368:Aged; D000438:Alcohols / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D003124:Colorimetry / Q000379:methods; D004950:Esterases / Q000097:blood* / Q000652:urine; D005260:Female; D006801:Humans; D007700:Kinetics; D008297:Male; D008875:Middle Aged; D010648:Phenylacetates; D012737:Sex Factors
D000438:Alcohols; D010648:Phenylacetates; D004950:Esterases; D002118:Calcium