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Molecular analysis of the Corynebacterium glutamicum gdh gene encoding glutamate dehydrogenase
Article first published online: 27 OCT 2006
DOI: 10.1111/j.1365-2958.1992.tb01474.x
1365-2958/asset/cover.gif?v=1&s=75254d9e3ae6a0f08085f34cc6e5bd65d8e3d52e "Molecular Microbiology")
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How to Cite
Börmann, E. R., Eikmanns, B. J. and Sahm, H. (1992), Molecular analysis of the Corynebacterium glutamicum gdh gene encoding glutamate dehydrogenase. Molecular Microbiology, 6: 317–326. doi: 10.1111/j.1365-2958.1992.tb01474.x
Publication History
- Issue published online: 27 OCT 2006
- Article first published online: 27 OCT 2006
- Received 23 August, 1991; revised and accepted 24 October, 1991.
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Summary
The Corynebacterium glutamicum gdh gene encoding NADP-dependent glutamate dehydrogenase (GDH) has been isolated by complementation of the Escherichia coli gdh mutant PA340. The gdh gene was subcloned into the E. coli/C. glutamicum shuttle vector pEK0 and introduced into C. glutamicum. Recombinant strains showed approximately eightfold higher specific GDH activity (15U mg protein-1) relative to the wild type (1.8U mg protein -1). Physiological studies with wild-type and recombinant C. glutamicum strains revealed no indication of significant regulation of gdh expression. The DNA sequence of 2082 bp, including the gdh gene, 5′-, and 3′-flanking regions, was determined. The structural gene consists of 1344 bp and codes for a polypeptide of 448 amino acid residues (Mr 49152) showing up to 53.6% identity with reported amino acid sequences of glutamate dehydrogenases from other organisms. Northern blot hybridization revealed a 1.65 kb mRNA transcript, indicating that the gdh gene of C. glutamicum is monocistronic. Transcription occurred from a G residue located 284bp upstream of the AUG considered to be the translational initiation codon.