HETEROLOGUS OVER-EXPRESSION AND IN-SILICO CHARACTERISATION OF A PUTATIVE 2-KETO-GLUCONATE DEHYDROGENASE FROM ARTHROBACTER NICOTINOVORANS pAO1

• Main Authors: Brandusa Cheorbeja, Madalina Bianca Bujder, Claudiu Arnautu, Marius Mihasan
• Format: Article
• Language: English
• Published: "Alexandru Ioan Cuza" University of Iași 2015-09-01
• Series: Analele Ştiinţifice Ale Universităţii Alexandru Ioan Cuza din Iași,Sectiunea II A : Genetica si Biologie Moleculara
• Online Access: http://www.gbm.bio.uaic.ro/index.php/gbm/article/view/1159

Arthrobacter nicotinovorans is a gram positive soil actinobacteria which is able to grow on nicotine contaminated soils due to the presence of a large plasmid - pAO1. It has been shown that pAO1 encodes not only the pathway for nicotine mineralization, but also newly described oxidative xylose-catabolic pathway. Part of the genes cluster encoding this last pathway is also gdh, a putative 2-keto-gluconate dehydrogenase with no experimentally shown function. Based on sequence homology, a 3D model of the GDH protein was generated. The protein over-expression was tested on two growth mediums, but was found to be satisfactory only on LB medium when using 0.1 mM IPTG. Using immobilized metal affinity chromatography, GDH was purified to homogeneity, but the yield was extremely low and did not allowed for further characterization of the protein.