Purification and characterization of L-aminoacylase from Pseudomonas maltophila B1

Mamoru Wakayama, Eiichi Shiiba, Kenji Sakai, Mitsuaki Moriguchi

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

The constitutive L-aminoacylase, which is used for optical resolution of DL-α-aminosuberic acid (DL-Asu), has been purified and characterized from Pseudomonas maltophila B1. The crude enzyme showed a specific activity of 0.062 units/mg for N-acetyl(Ac)-L-Asu. This value is very high compared with those from Aspergillus melleus, porcine kidney, and Bacillus stearothermophilus. Molecular masses of 108 kDa for the native enzyme and 50 kDa for the subunit were determined, indicating a dimer. The enzyme activity was optimal at pH 8.0 and at 55°C. The enzyme hydrolyzed N-acyl derivatives of various neutral L-amino acids and acidic L-amino acids, L-glutamate and L-Asu. The enzyme also had dipeptidase activity. The Km values for N-Ac-L-alanine and N-Ac-DL-Asu were determined at 2.32 and 12.7 mM, respectively. The apoenzyme was activated using Zn2+, Ca2+, and Co2+. Glyoxylate, DL-lactate, phenylboronic acid (PBA), butaneboronic acid (BBA), diethylpyrocarbonate (DEP), and phenylglyoxal (PGO) inhibited enzyme activity.

Original languageEnglish
Pages (from-to)278-282
Number of pages5
JournalJournal of Fermentation and Bioengineering
Volume85
Issue number3
DOIs
Publication statusPublished - Jan 1 1998
Externally publishedYes

Fingerprint

Pseudomonas
Purification
Enzymes
Acids
Enzyme activity
Amino acids
Phenylglyoxal
Diethyl Pyrocarbonate
Apoenzymes
Acidic Amino Acids
Aspergillus
Molecular mass
Bacilli
Dimers
Neutral Amino Acids
Geobacillus stearothermophilus
Glutamic Acid
Lactic Acid
Derivatives
Amino Acids

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Purification and characterization of L-aminoacylase from Pseudomonas maltophila B1. / Wakayama, Mamoru; Shiiba, Eiichi; Sakai, Kenji; Moriguchi, Mitsuaki.

In: Journal of Fermentation and Bioengineering, Vol. 85, No. 3, 01.01.1998, p. 278-282.

Research output: Contribution to journalArticle

Wakayama, Mamoru ; Shiiba, Eiichi ; Sakai, Kenji ; Moriguchi, Mitsuaki. / Purification and characterization of L-aminoacylase from Pseudomonas maltophila B1. In: Journal of Fermentation and Bioengineering. 1998 ; Vol. 85, No. 3. pp. 278-282.
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