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 1 1996
Externally publishedYes

Fingerprint

Diethyl Pyrocarbonate
Chemical modification
Pseudomonas
Histidine
Enzymes
Hydroxylamine
Enzyme activity
Glutamic Acid
Catalytic Domain
Sodium
Kinetics
Substrates
N-acyl-D-glutamate deacylase

All Science Journal Classification (ASJC) codes

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

Cite this

Chemical Modification of Histidine Residue of N-Acyl-D-Glutamate Amidohydrolase from Pseudomonas sp. 5f-1; / Wakayama, Mamoru; Tsutsumi, Tetsuo; Yada, Harutaka; Sakai, Kenji; Moriguchi, Mitsuaki.

In: Bioscience, Biotechnology and Biochemistry, Vol. 60, No. 4, 01.01.1996, p. 650-653.

Research output: Contribution to journalArticle

Wakayama, Mamoru ; Tsutsumi, Tetsuo ; Yada, Harutaka ; Sakai, Kenji ; Moriguchi, Mitsuaki. / Chemical Modification of Histidine Residue of N-Acyl-D-Glutamate Amidohydrolase from Pseudomonas sp. 5f-1;. In: Bioscience, Biotechnology and Biochemistry. 1996 ; Vol. 60, No. 4. pp. 650-653.
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