Molecular analysis of L-haloacid dehalogenase from Arthrobacter ureafaciens sp SI

Abstract

Using enrichment technique a bacterium was isolated from soil showing the ability to grow on 20 mM 2, 2-dichloropropionate (2, 2-DCP) whether in solid or in liquid minimal media. Further characterization was carried out using 16S rRNA analysis. The partial 16S rRNA sequence (1450bp) was BLASTn in the NCBI database resulted in 95% identity to Arthrobacter ureafaciens (Accession number. FN433020.1). Therefore, this isolate was designated as Arthrobacter ureafaciens S1. In this study, we focused on the dehalogenase gene identification by PCR amplification using genomic DNA from Arthrobacter strain S1. Two kinds of primers DehA (reverse&forward) and DehB (reverse&forward) were used for dehalogenase gene amplification. The PCR results showed 1300 bp and 700 bp using DehA (reverse&forward) and DehB (reverse&forward), respectively. Moreover, the partial gene sequence was assembled, analyzed and compared with Gen Bank database in NCBI (National Center for Biotechnology Information) and converted into amino acids sequence. Using BLASTp tools, analysis showed distance relationship with L-2-haloacid sequence amino sequence that belonged to group II of a-halocarboxylic acid (aHA) dehalogenase. In conclusion, current investigations suggested that there were more than one dehalogenases were amplified and these dehalogenases maybe a novel based on the size of the amplified band patterns compared to the previously reported. However, further investigations need to be carried out.