Combinatorial approach for screening and assessment of multiple therapeutic enzymes from marine isolate Pseudomonas aeruginosa AR01.

Abstract

Industrialization and modernization have led to humans being more susceptible to diseases. Therapeutic enzymes from traditional earthbound bacterial origin result have less therapeutic value. Hence, the hunt for a novel source of enzymes is indispensable. Twenty different marine bacterial strains were isolated from mangrove soil around S. P. Pattinum, Tamilnadu, India. From repeated qualitative and quantitative experiments, the study results were that, out of twenty bacterial isolates, only one Gram-negative bacterium was 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 medium 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 that the therapeutic enzymes from P. aeruginosa AR01 may find a significant role in medical applications. The in vitro cytotoxic study reveals that the anticancer enzyme from P. aeruginosa is considerably effective with an IC50 value of 12 μg mL−1 against K-562 cell line. Colony PCR was performed for the detection of specific therapeutic enzyme-coding genes in the genome of P. aeruginosa AR01. PCR results confirm that P. aeruginosa AR01 possesses nucleotide regions for corresponding therapeutic enzymes in its gene cluster. BLASTN and BLASTX analyses of the partial nucleotide sequences of therapeutic enzymes were deposited in GenBank. The results appear so promising that Pseudomonas aeruginosa AR01 may be a potent candidate for medical biotechnology.