Nucleotide sequence, cloning, and overexpression of the d-threonine dehydrogenase gene from Pseudomonas cruciviae |
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| Authors: | Makoto Ashiuchi Kanoktip Packdibamrung Toshiyuki Miyaji Shinji Nagata Haruo Misono |
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| Institution: | Research Institute of Molecular Genetics, Kochi University, Nankoku, Kochi 783-8502, Japan;Department of Bioresources Science, Kochi University, Nankoku, Kochi 783-8502, Japan |
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| Abstract: | d-Threonine dehydrogenase (EC 1.1.1) catalyses the oxidation of the 3-hydroxyl group of d-threonine. The nucleotide sequence of the structural gene, dtdS, for this enzyme from Pseudomonas cruciviae IFO 12047 was determined. The dtdS gene encodes a 292 amino acid polypeptide. The enzyme was overproduced in Escherichia coli cells; the activity was found in cell extracts of the clone. The enzyme showed high sequence similarity to 3-hydroxyisobutyrate dehydrogenases. This is the first example showing the primary structure of an enzyme catalysing the NADP+-dependent dehydrogenation of d-threo-3-hydroxyamino acids. |
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| Keywords: | d-Threonine dehydrogenase gene Pseudomonas cruciviae 3-Hydroxyacid dehydrogenase superfamily Nucleotide sequence Amino acid sequence |
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