標題: Discovery of a novel N-iminylamidase activity: substrate specificity, chemico selectivity and catalytic mechanism
作者: Huang, CY
Yang, YS
生醫工程研究所
Institute of Biomedical Engineering
關鍵字: N-iminylamidase;imidase;hydantoinase;dihydropyrimidinase
公開日期: 1-Mar-2005
摘要: 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, C-13 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. (c) 2004 Elsevier Inc. All rights reserved.
URI: http://dx.doi.org/10.1016/j.pep.2004.12.008
http://hdl.handle.net/11536/13943
ISSN: 1046-5928
DOI: 10.1016/j.pep.2004.12.008
期刊: PROTEIN EXPRESSION AND PURIFICATION
Volume: 40
Issue: 1
起始頁: 203
結束頁: 211
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