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

研究成果: ジャーナルへの寄稿記事

18 引用 (Scopus)

抄録

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

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Alcaligenes
D-Aspartic Acid
Purification
Enzymes
p-Chloromercuribenzoic Acid
Iodoacetic Acid
Amino Acids
Sulfhydryl Reagents
Ethylmaleimide
Dithiothreitol
Divalent Cations
Isoelectric Point
Molecular mass
Electrophoresis
Sodium Dodecyl Sulfate
Polyacrylamide Gel Electrophoresis
Glutamic Acid
Amino Acid Sequence
Monomers
Derivatives

All Science Journal Classification (ASJC) codes

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

これを引用

Purification and Characterization of Novel N-Acyl-D-aspartate Amidohydrolase from Alcaligenes xylosoxydans subsp. Xylosoxydans A-6. / Moriguchi, Mitsuaki; Sakai, Kenji; Katsuno, Yutaka; Maki, Tetsuyoshi; Wakayama, Mamoru.

:: Bioscience, biotechnology, and biochemistry, 巻 57, 番号 7, 01.01.1993, p. 1145-1148.

研究成果: ジャーナルへの寄稿記事

Moriguchi, Mitsuaki ; Sakai, Kenji ; Katsuno, Yutaka ; Maki, Tetsuyoshi ; Wakayama, Mamoru. / Purification and Characterization of Novel N-Acyl-D-aspartate Amidohydrolase from Alcaligenes xylosoxydans subsp. Xylosoxydans A-6. :: Bioscience, biotechnology, and biochemistry. 1993 ; 巻 57, 番号 7. pp. 1145-1148.
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abstract = "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.",
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AB - 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.

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