Purification and Characterization of Novel N-Acyl-D-aspartate Amidohydrolase from Alcaligenes xylosoxydans subsp. Xylosoxydans A-6

Mitsuaki Moriguchi, Kenji Sakai, Yutaka Katsuno, Tetsuyoshi Maki, Mamoru Wakayama

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Alcaligenes xylosoxydans subsp. xylosoxydans A-6 (Alcaligenes A-6) produced N-acyl-D-aspartate amidohydrolase (D-AAase) in the presence of N-acetyl-D-aspartate as an inducer. The enzyme was purified to homogeneity. The enzyme had a molecular mass of 56 kDa and was shown by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) to be a monomer. The isoelectric point was 4.8. The enzyme had maximal activity at pH 7.5 to 8.0 and 50°C, and was stable at pH 8.0 and up to 45°C. N-Formyl (Km=12.5mM), N-acetyl (Km=2.52mM), N-propionyl (Km=0.194mM), N-butyryl (Km=0.033mM), and N-glycyl (Km=1.11mM) derivatives of D-aspartate were hydrolyzed, but N-carbobenzoyl-D-aspartate, N-acetyl-L-aspartate, and N-acetyl-D-glutamate were not substrates. The enzyme was inhibited by both divalent cations (Hg2+, Ni2+, Cu2+) and thiol reagents (N-ethylmaleimide, iodoacetic acid, dithiothreitol, and p-chloromercuribenzoic acid). The N-terminal amino acid sequence and amino acid composition were analyzed.

元の言語英語
ページ(範囲)1145-1148
ページ数4
ジャーナルBioscience, biotechnology, and biochemistry
57
発行部数7
DOI
出版物ステータス出版済み - 1 1 1993
外部発表Yes

All Science Journal Classification (ASJC) codes

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

フィンガープリント Purification and Characterization of Novel N-Acyl-D-aspartate Amidohydrolase from Alcaligenes xylosoxydans subsp. Xylosoxydans A-6' の研究トピックを掘り下げます。これらはともに一意のフィンガープリントを構成します。

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