Combinatorial approach for screening and assessment of multiple therapeutic enzymes from marine isolate pseudomonas aeruginosa ar01

Abstract

Industrialization and modernization led humans more susceptible to diseases. The therapeutic enzymes from the traditional earthbound bacterial origin result in less therapeutic value. Hence, the hunt for a novel source of enzymes is much indispensable. Twenty different marine bacterial strains were isolated from mangrove soil around S. P. Pattinum, Tamilnadu, India. From the repeated qualitative and quantitative experiments, the study results were, out of twenty bacterial isolates only one, gram-negative bacterium exhibits positive for multiple therapeutic enzymes such as asparaginase, glutaminase, uricase and collagenase. Based on its 99% 16S rRNA sequence similarity with Pseudomonas aeruginosa, the isolate was designated as Pseudomonas aeruginosa AR01. Modified minimal media amended with asparagine results in a simple and cost-effective, one-pot production medium for enhanced production and easy purification of all therapeutic enzymes. The biochemical studies imply the therapeutic enzymes from P. aeruginosa AR01 may find a significant role in medical applications. The in-vitro cytotoxic study reveals the anticancer enzymes from P. aeruginosa is considerably effective with an IC50 value of 12 µg/mL against K-562 cell line. Colony PCR was performed for the detection of specific therapeutic enzymes coding gene in the genome of Pseudomonas. aeruginosa AR01. PCR results confirm that Pseudomonas aeruginosa AR01 possess nucleotide regions for corresponding therapeutic enzymes in its gene cluster. BLASTN and BLASTX analysis of the partial nucleotide sequences of therapeutic enzymes were deposited in GenBank. The results appear so promising that Pseudomonas aeruginosa AR01, may employ as a potent candidate for the medical biotechnology.Keywords: Marine bacteria, Therapeutic enzymes, Pseudomonas aeruginosa, Colony PCR, Cytotoxicity, Nucleotide sequences newline