Cloning and Overexpression of the 3-Hydroxyisobutyrate Dehydrogenase Gene fromPseudomonas putidaE23 | Zendy

Emran Kabir Chowdhury | Zendy; Yuka AKAISHI | Zendy; Shinji Nagata | Zendy; Haruo Misono | Zendy

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Cloning and Overexpression of the 3-Hydroxyisobutyrate Dehydrogenase Gene fromPseudomonas putidaE23

Author(s) -

Emran Kabir Chowdhury,

Yuka AKAISHI,

Shinji Nagata,

Haruo Misono

Publication year - 2003

Publication title -

bioscience biotechnology and biochemistry

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.509

H-Index - 116

eISSN - 1347-6947

pISSN - 0916-8451

DOI - 10.1271/bbb.67.438

Subject(s) - pseudomonas putida , dehydrogenase , escherichia coli , biochemistry , biology , gene , microbiology and biotechnology , recombinant dna , enzyme , gene cluster

The structural gene for NAD+-dependent 3-hydroxyisobutyrate dehydrogenase (EC 1.1.1.31) from Pseudomonas putida E23 was cloned in Escherichia coli cells to obtain a large amount of the enzyme and its nucleotides were sequenced to study its structural relationship with other proteins. The gene encoded a polypeptide containing 295 amino acid residues and was in a cluster with the gene for methylmalonate semialdehyde dehydrogenase. Transformed E. coli cells overproduced 3-hydroxyisobutyrate dehydrogenase, and the recombinant enzyme was purified to homogeneity with a high yield. Lysine and asparagine residues, which are important in catalysis of the 3-hydroxyacid dehydrogenase family, are conserved in this enzyme.

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