DETECTION AND ISOLATION OF RESTRICTION ENDONUCLEASE LIKE TOPOISOMERASE FROM THERMOPHILIC BACILLUS SUBTILUS STRAIN RC1

Aims: Detection and isolation of restriction endonucleases like topoisomerase   Thermophilic Bacillus subtilus strain RC1. Methods and results: Topoisomerase like enzyme activity is detected from thermophilc Bacilus sp. The isolated enzyme is partially characterized. The molecular weight of the enzyme is ~ 33 KDa and the optimum temperature for the activity is 37 ⁰C. The crude and gel purified enzyme activity were performed. The enzyme could be novel one, as the isolate is from hot water ecosystem. The restriction pattern of the new isolated enzyme is dissimilar to the commercially available enzymes. The enzyme retained the activity at a storage temperature of 4 ⁰C about one to two months. Magnesium chloride is found to be essential for the activity. The enzyme activity pattern is like topoisomrase. Activity is enhance in presence of ATP. The further characterization of topo like enzyme is in contined.Conclusion: The endonuclease type of topoisomerase enzyme is detected and isolated from thermophilic Bacillus sp.The restriction pattern studies indicate that the enzyme cuts standard plasmid DNA. Significant impact of the study: The restriction enzyme which is isolated from thermophilic Bacillus sp. and this microorganism was isolated from hot spring ecosystem. So enzyme might be new restriction endonuclease and there are few reports from thermophilic microorganisms. Restriction endonucleases with specific site of action will be useful in genetic engineering studies.