Mechanism of biochemical action of substituted 4-methylbenzopyran-2-ones. Part 6: hydrolysis of 7,8-diacetoxy-4-methylcoumarin by a novel deacetylase in rat liver microsomes--a simple method for assay and characterisation.
The existence of a novel microsomal deacetylase in rat liver catalysing deacetylation of diacetoxy 4-methylcoumarins has been reported. A simple method is outlined for the enzyme assay based upon the quantification of the dihydroxy derivative by measuring the UV absorption of its complex with ADP and Fe3+ at 600 nm. The enzyme can be routinely assayed using 7,8-diacetoxy-4-methylcoumarin (DAMC) as the substrate and demonstrated hyperbolic kinetics and yielded Km and vmax values of 1250 microM and 500 units, respectively. The pH optima was found to be 7.5 for the enzyme. No DAMC deacetylase activity was found in hepatic cytosol and the enzyme activity was not discernible in extrahepatic tissues.
Raj
H R
HR
Department of Biochemistry, VP Chest Institute, University of Delhi, India.
Parmar
V S
VS
Jain
S C
SC
Goel
S
S
Tyagi
Y K
YK
Sharma
S K
SK
Olsen
C E
CE
Wengel
J
J
D000115:Acetylesterase / Q000378:metabolism*; D000818:Animals; D001578:Benzopyrans / Q000737:chemistry / Q000378:metabolism*; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D003374:Coumarins / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D051381:Rats; D017208:Rats, Wistar; D013379:Substrate Specificity
C114369:7,8-diacetoxy-4-methylcoumarin; D001578:Benzopyrans; D003374:Coumarins; C417192:7,8-diacetoxy-4-methylcoumarin deacetylase; D002265:Carboxylic Ester Hydrolases; D000115:Acetylesterase