Gene cloning and characterization of a deaminase from the 4-amino-3-hydroxybenzoate-assimilating Bordetella sp. strain 10d |
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Authors: | Shinji Takenaka Tomomi Sato Jyun Koshiya Shuichiro Murakami & Kenji Aoki |
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Institution: | Department of Applied Biological Chemistry, Graduate School of Agricultural Science, Kobe University, Kobe, Japan;and;Department of Nutritional Management, Faculty of Nutritional Science, Sagami Women's University, Sagamihara, Japan |
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Abstract: | The 4-amino-3-hydroxybenzoate-assimilating Bordetella sp. strain 10d produces a deaminase that catalyzes the deamination of 2-amino-5-carboxymuconic 6-semialdehyde. A gene encoding the deaminase, ahdB , was cloned and expressed in Escherichia coli; ahdB is located downstream from the previously reported genes encoding 4-amino-3-hydroxybenzoate 2,3-dioxygenase ( ahdA ) and a LysR-type regulator. The deduced amino acid sequence of ahdB shows 30–33% identity to those of previously reported 2-aminomuconate deaminases. We identified a region (RAGDFLXVSG) conserved in AhdB and three other deaminases. The recombinant enzyme AhdB was purified to homogeneity. After a coupled enzyme assay with purified AhdA, AhdB, and the substrate 4-amino-3-hydroxybenzoate, the final product, formed by the action of AhdA, AhdB, and by nonenzymatic decarboxylation, was identified by HPLC, MS, and 1H-nuclear magnetic resonance analyses as 2-hydroxymuconic 6-semialdehyde. |
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Keywords: | Bordetella 2-aminophenol derivative 4-amino-3-hydroxybenzoate 2-amino-5-carboxymuconic 6-semialdehyde 2-amino-5-carboxymuconic 6-semialdehyde deaminase 2-aminomuconate deaminase |
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