Cloning Glucose Dehydrogenase Gene of Gluconobacter suboxydans through TAIL- PCR and Its Bioinformatics Analysis

PQQ-dependent glucose dehydrogenase, which catalyzes D-glucose to product gluconic acid, is an important enzyme in the production of gluconic acid by fermentation or enzymatic method. Primers were designed based on the conserved region of PQQ attaching sites, and the whole gdh gene was obtain by TAIL-PCR. Then the gene was sequenced and analyzed by bioinformatics methods. The sequence analysis suggested that the coding region consisted of the 2268 nucleotides which encoded 755 amino acids. The deduced amino acid sequence showed high level similarity with Gluconobacter oxydans. The molecular weight of the protein was 81.72kD and the isoelectric point was 5.14.The bioinformatics analysis suggested that its second structure consisted of 18.41% alpha helix, 16.16% extended strand and 64.43% random coil and its N-terminal contained five transmembrane domains which located between sites 1 and 140 area. TAIL-PCR provided a simple and efficient method for the cloning of the unknown genes. The bioinformatics analysis of GDH provided a foundation for further investigation on the