Aldoxime Dehydratase Co-existing with Nitrile Hydratase and Amidase in the Iron-Type Nitrile Hydratase-Producer Rhodococcus sp. N-771 (ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)

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概要

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• We identified an aldoxime dehydratase (Oxd) gene in the 5'-flanking region of the nitrile hydratase-amidase gene cluster in the photoreactive iron-type nitrile hydratase-producer, Rhodococcus sp. N-771. The enzyme showed 96.3%, 77.6%, and 30.4% identities with the Oxds of Rhodococcus globerulus A-4, Pseudomonas chlororaphis B23, and Bacillus sp. OxB-1, respectively. The enzyme was expressed in Escherichia coli under the control of the lac- or T7 promoters in its intact and His_6-tagged forms, purified, and characterized. The enzyme had heme b as a prosthetic group, catalyzed a stoichiometric dehydration of aldoxime into nitrile, and exhibited the highest activity at neutral pH and at around 30℃ similar to the known Oxd from Bacillus sp. OxB-1. The activity was enhanced by reducing agents, such as Na_2S, Na_2S_2O_4, 2-mercaptoethanol, and L-cysteine and supplementary additions of electron acceptors such as flavins, sulfite ion, and vitamin K_3. The effect of various chemicals on the enzyme activity was different in the presence and absence of the reducing reagent, Na_2S. The enzyme preferentially acts on aliphatic-type substrates and the substrate specificity of the enzyme coincides with that reported for nitrile hydratase produced by the strain.

• 社団法人日本生物工学会の論文
• 2004-04-25

著者

• Asano Yasuhisa

  Biotechnology Research Center, Toyama Prefectural University

• Xie Sheng-xue

  Biotechnology Research Center Faculty Of Engineering Toyama Prefectural University

• Asano Y

  Department Of Immunology And Host Defenses Ehime University Graduate School Of Medicine

• Asano Y

  Toyama Prefectural Univ. Toyama Jpn

• Kato Y

  Biotechnology Research Center Faculty Of Engineering Toyama Prefectural University

• KATO YASUO

  Biotechnology Research Center, Faculty of Engineering, Toyama Prefectural University

• YOSHIDA SATOSHI

  Biotechnology Research Center, Faculty of Engineering, Toyama Prefectural University

• Asano Yasuhisa

  Biotechnology Res. Center And Dep. Of Biotechnology Toyama Prefectural Univ.

• Kato Yasuo

  Biotechnology Research Center And Department Of Biotechnology Toyama Prefectural University

• Yoshida Satoshi

  Biotechnology Research Center Faculty Of Engineering Toyama Prefectural University

• Xie Seng-Xue

  Biotechnology Research Center, Faculty of Engineering, Toyama Prefectural University

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