Discovery of a novel N-iminylamidase activity: substrate specificity, chemicoselectivity and catalytic mechanism.
Enzymatic hydrolysis of the N-iminylamide was investigated in this study. An enzyme possessing N-iminylamidase activity from pig liver was purified to electrophoretic homogeneity. This enzyme was also active, however, with imides and appears to be identical to pig liver imidase. The identification was confirmed by copurification of enzyme activities and by specificities of typical substrates of mammalian imidase, such as phthalimide, dihydrouracil, and maleimide. The hydrolysis of 3-iminoisoindolinone was further analyzed by HPLC, (13)C NMR spectrometry, and LC-MS measurements to determine its chemicoselectivity. All data indicated that this enzyme chemicoselectively catalyzed the hydrolysis of the N-iminylamide to produce the compound bearing the diamine and carboxylate group. The pH profiles of this enzyme suggest that one of the protons of 3-iminoisoindolinone was important to promote the ring-opening process of this substrate. These results constituted a first study on the enzymatic hydrolysis of compounds bearing the N-iminylamide functional group.
Huang
Cheng-Yang
CY
Institute of Biochemical Engineering, College of Biological Science and Technology, National Chiao Tung University, Hsinchu, Taiwan, ROC.
Yang
Yuh-Shyong
YS
D000581:Amidohydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D000818:Animals; D006863:Hydrogen-Ion Concentration; D008099:Liver / Q000201:enzymology; D008970:Molecular Weight; D013379:Substrate Specificity; D013552:Swine
D000581:Amidohydrolases; C019665:amidase; C020047:dihydropyrimidinase