Abstract: ATP sulfurylase (ATPS) is widely distributed in all living organisms. Several different physiological roles have been proposed for ATPS in different species, including sulfate assimilation, sulfate reduction and pyrophosphate recycling. Also, ATP sulfurylase has many different industrial and laboratory applications. The aim of this study was to clone and express the gene that producing the recombinant ATPS protein from an Iranian strain of Geobacillus. After Isolation and identification of Geobacillus kaustophilus strain, DNA genomic was extracted. ATPS gene was amplified from genomic DNA by using a couple of specific primers for interested gene. PCR product of ATPS gene was observed as an 1188bp band on agarose gel. Then the PCR product was purified and cloned into the cloning vector. The ATPS band was sequenced after cloning and result of homology search in the NCBI database confirmed that the cloned gene was ATPS. The ATPS gene was subcloned in expression pET28a plasmid. Expression of recombinant ATPS protein in E. Coli BL21 (DE3) was analyzed using SDS-PAGE gel. Analysis of expressed ATPS protein on SDS-PAGE gel revealed a band at 47.5 KD. Using ATP luminescence method for measuring enzymatic activity of the protein showed that the recombinant protein is active. This is the first study on cloning, expression and enzymatic activity of the ATPS gene from the Geobacillus kaustophilus bacteria.

Keywords: ATP sulfurylase, pyrophosphate, Geobacillus kaustophilus, recombinant protein and ATP luminescence

Full-Text [PDF 444 kb]   (6788 Downloads)