Molecular characterization of a novel bacterial aryl acylamidase belonging to the amidase signature enzyme family.

Hyeok Jin Ko, Eun Woo Lee, Won Gi Bang, Cheol-Koo Lee, Kyoung Heon Kim, In-Geol Choi

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

In seeking aryl acylamidase (EC 3.5.1.13) acting on an amide bond in p-acetaminophenol (Tylenol), we identified a novel gene encoding 496 residues of a protein. The gene revealed a conserved amidase signature region with a canonical catalytic triad. The gene was expressed in E. coli and characterized for its biochemical properties. The optimum pH and temperature for the activity on p-acetaminophenol were 10 and 37 degrees C, respectively. The half-life of enzyme activity at 37 degrees C was 192 h and 90% of its activity remained after 3 h incubation at 40 degrees C. Divalent metals was found to inhibit the activity of enzyme. The K (m) values for various aryl acylamides such as 4-nitroacetanilide, p-acetaminophenol, phenacetin, 4-chloroacetanilide and acetanilide were 0.10, 0.32, 0.83, 1.9 and 19 mM, respectively. The reverse reaction activity (amide synthesis) was also examined using various chain lengths (C(1) approximately C(4) and C(10)) of carboxylic donors and aniline as substrates. These kinetic parameters and substrate specificity in forward and reverse reaction indicated that the aryl acylamidase in this study has a preference for aryl substrate having polar functional groups and hydrophobic carboxylic donors.

Original languageEnglish
Pages (from-to)485-492
Number of pages8
JournalMolecules and Cells
Volume29
Issue number5
DOIs
Publication statusPublished - 2010 May 1

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aryl acylamidase
amidase
Amides
Enzymes
Phenacetin
Genes
Acetaminophen
Substrate Specificity
Half-Life
Metals
Escherichia coli
Temperature
Proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Molecular characterization of a novel bacterial aryl acylamidase belonging to the amidase signature enzyme family. / Ko, Hyeok Jin; Lee, Eun Woo; Bang, Won Gi; Lee, Cheol-Koo; Kim, Kyoung Heon; Choi, In-Geol.

In: Molecules and Cells, Vol. 29, No. 5, 01.05.2010, p. 485-492.

Research output: Contribution to journalArticle

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