Discovery of a novel N-iminylamidase activity: Substrate specificity, chemicoselectivity and catalytic mechanism

Cheng Yang Huang, Yuh-Shyong Yang*

*Corresponding author for this work

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

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, 13C 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.

Original languageEnglish
Pages (from-to)203-211
Number of pages9
JournalProtein Expression and Purification
Volume40
Issue number1
DOIs
StatePublished - 1 Jan 2005

Keywords

  • Dihydropyrimidinase
  • Hydantoinase
  • Imidase
  • N-iminylamidase

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