Comparative investigation of 16 xenobiotic metabolizing N-acetyltransferase (NAT) homologs from bacteria. Bacterial NAT genes

Amplification and sequencing (automated Sanger method) of 16 genes encoding homologs of arylamine N-acetyltransferase (NAT) in 11 bacterial species, including 6 low-GC gram-positive Firmicutes from the order of Bacillales (microbiologically identified as Bacillus amyloliquefaciens subsp. plantarum, Bacillus cereus, Bacillus fordii, Bacillus licheniformis, Bacillus safensis and Brevibacillus borstelensis), 4 high-GC gram-positive Actinobacteria from the orders of Corynebacteriales (Tsukamurella paurometabola) and Streptomycetales (identified as Streptomyces griseus, Streptomyces sulphureus and Streptomyces venezuelae), and 1 gram-negative Alphaproteobacterium from the order of Rhizobiales (Afipia birgiae). The PCR-amplified products were cloned into pET28b(+) expression vector for the purpose of recombinant protein expression, purification and subsequent enzymatic analyses. This work was part of a comparative functional investigation into bacterial NAT homologs in the framework of a collaboration between Democritus University of Thrace (Department of Molecular Biology and Genetics, Alexandroupolis, Greece) and ELTE Eötvös Loránd University (Department of Microbiology, Budapest, Hungary) funded by a Joint Research & Technology Program between Greece and Hungary, co-financed by Greece, Hungary and the European Union (European Regional Development Fund-ERDF).