Cloning, characterization and DNA sequencing of the gene encoding the Mr50000 quinoprotein glucose dehydrogenase fromAcinetobacter calcoaceticus

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Abstract
Recently we described the cloning of the gene coding for a Mr 87000 glucose dehydrogenase (GDH-A) fromAcinetobacter calcoaceticus. In this report we describe the cloning of a gene coding for a second GDH (GDH-B) with a Mr of 50000 from the same organism. This gene was isolated using a 20-mer synthetic oligonucleotide, derived from the N-terminal amino acid sequence of purified GDH-B as a probe to screen a genomic bank. From the DNA sequence of thegdhB gene, a protein can be derived of Mr 52772 with a 24 amino acid signal peptide which is removed, resulting in the mature protein with a Mr 50231. In vitro transcription-translation of thegdhB clone shows the mature and the precursor protien. The derived amino acid sequence has no obvious homology with GDH-A ofA. calcoaceticus. We show that disaccharides are specific GDH-B substrates and that 2-deoxyglucose is specific for GDH-A.