Purification and Properties of an Aminopeptidase from a Protamine-degrading Marine Bacterium

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概要

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• A protamine-degrading marine bacterium was isolated from marine soil and identified as Aeromonas salmonicida subsp. based on its taxonomical characteristics. An alanine-specific aminopeptidase, called aminopeptidase K, from an extract of the strain was purified and characterized. The aminopeptidase K was purified about 80-fold by fractionation with ammonium sulfate and column chromatography on QA-52 cellulose, Phenyl Superose and Superose 12. The purified enzyme is composed of 6 subunits of 86 kDa with a molecular mass of 520 kDa according to gel filtration and SDS-PAGE. The N-terminal sequence of the enzyme was H・Gly-Gln-Gln-Pro-Gln-Ile-Lys-Try-Tyr-His-Asp-Tyr-Asp-Ala-Pro-Asp-Tyr-Tyr-Ile-Thr-. It is inhibited by monoiodoacetate, N-ethylmaleimide, and puromycin. The Michaelis constant (K_m) and the maximal rate of hydrolysis (V_<max>) were, respectively, 0.28 mM and 49.4 μmol/min/mg for the L-Ala-β-naphthylamide substrate. The optimum pH and optimum temperature were 6.5 and 45℃, respectively. The purified enzyme was highly specific to L-Ala-β-naphthylamide.

• 社団法人日本農芸化学会の論文
• 1997-07-23

著者

• Obata H

  Department Of Biotechnology Faculty Of Engineering Kansai University

• Obata Hitoshi

  Department Of Biotechnology Faculty Of Engineering Kansai University

• Sugiyama Atsushi

  Department of Pharmacology, School of Medicine, Toho University, Japan

• Kawahara Hidehisa

  Department of Biotechnology, Faculty of Engineering, Kansai University

• Hidehisa Kawahara

  Department Of Biotechnology Faculty Of Engineering Kansai University:high Technology Research Center

• Kawahara Hidehisa

  Department Of Biological Function And Genetic Resources Science Faculty Of Agriculture Okayama Unive

• Muramatsu Tsuyoshi

  Faculty Of Fisheries Nagasaki University

• Obata Hitoshi

  Graduate School Of Bioresources Mie Univ.

• Obata Hitoshi

  Department Of Agricultural Chemistry Faculty Of Bioresources Mie University Department Of Biotechnol

• Sugiyama Atsushi

  Department Of Pharmacology Yamanashi Medical University

• Sugiyama A

  Department Of Biotechnology Faculty Of Engineering Kansai University

• Kawaguchi H

  Department Of Orthopaedic Surgery Graduate School Of Medicine The University Of Tokyo

• Sugiyama Atsushi

  Department Of Chemistry College Of Science Rikkyo University

• Kawaguchi Hiroshi

  Department Of Orthopaedic Surgery The University Of Tokyo

• Sugiyama Atsushi

  Department of Biotechnology, Faculty of Engineering, Kansai University

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