Cloning, characterization and DNA sequencing of the gene encoding the Mr 50,000 quinoprotein glucose dehydrogenase from Acinetobacter calcoaceticus.
Recently we described the cloning of the gene coding for a Mr 87,000 glucose dehydrogenase (GDH-A) from Acinetobacter calcoaceticus. In this report we describe the cloning of a gene coding for a second GDH (GDH-B) with a Mr of 50,000 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 the gdhB gene, a protein can be derived of Mr 52,772 with a 24 amino acid signal peptide which is removed, resulting in the mature protein with a Mr 50,231. In vitro transcription-translation of the gdhB clone shows the mature and the precursor protein. The derived amino acid sequence has no obvious homology with GDH-A of A. calcoaceticus. We show that disaccharides are specific GDH-B substrates and that 2-deoxyglucose is specific for GDH-A.
Cleton-Jansen
A M
AM
Laboratory of Molecular Genetics, University of Leiden, The Netherlands.
Goosen
N
N
Vink
K
K
van de Putte
P
P
D000150:Acinetobacter / Q000201:enzymology / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D002237:Carbohydrate Dehydrogenases / Q000235:genetics*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D005948:Glucose Dehydrogenases / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009154:Mutation; D015183:Restriction Mapping
D004269:DNA, Bacterial; D002237:Carbohydrate Dehydrogenases; D005948:Glucose Dehydrogenases; C033955:glucose dehydrogenase (pyrroloquinoline-quinone)