Chemical Modification of Histidine Residue of N-Acyl-D-Glutamate Amidohydrolase from Pseudomonas sp. 5f-1;

Mamoru Wakayama, Tetsuo Tsutsumi, Harutaka Yada, Kenji Sakai, Mitsuaki Moriguchi

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

N-Acyl-D-glutamate amidohydrolase (D-AGase) from Pseudomonas sp. 5f-1 was inactivated by diethyl pyrocarbonate (DEP). The chemical modification by DEP showed a difference spectrum at 246 nm due to the N-carbethoxyhistidine residue. Removal of the carbethoxy group from inactivated enzyme with hydroxylamine restored enzyme activity. The inactivation by DEP proceeded with pseudo-first-order kinetics, and was protected in the presence of the substrate N-acetyl-D-glutamate (Glu), or the competitive inhibitor sodium α-ketoglutarate (α-KGA). These results suggest the presence of an essential histidine residue at or near of the active site of the enzyme.

Original languageEnglish
Pages (from-to)650-653
Number of pages4
JournalBioscience, Biotechnology and Biochemistry
Volume60
Issue number4
DOIs
Publication statusPublished - Jan 1996
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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