Abstract:

Nitroreductases (NTR) are members of the NAD(P)H/FMN oxidoreductase that exists widely in bacteria. NTRs have raised a great interest due to their potential in biomedicine, especially in prodrug activation for chemotherapeutic cancer treatments, in bioremediation and enzyme-based biosensors for nitro-sensitive compounds. Thermophilic Bacillus species isolated from hot water springs in Turkey were identified by 16S rRNA gene sequence analysis. The previously identified species, Bacillus paralicheniformis (5NK) and Bacillus paralicheniformis (FMB2) with sequence similarity of 97% and 99.65, respectively, were used. They were all found to possess the putative NADPH nitroreductase genes. Genomic DNAs for isolation of nitroreductase genes were isolated by conventional phenol-chloroform extraction method. The nitroreductase genes from thermophilic Bacillus species were amplified by PCR, using the designed three primer pairs. The corresponding DNA fragments were fractionated by agarose gel electrophoresis and PCR products were purified and concentrated for cloning and DNA sequencing.

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