Purification, Kinetic Characterization and Anti-Proliferative Activity of A Therapeutic Protein from Marine Sources

Rahamat Unissa, Merugula Sowmya, Yadla Sagarika, Veeravarapu Divya, Mundrathi Anusha, Ashwini Kumar Chauhan, Sowmya konakanchi and Sindhu Chowdary

Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Malla Reddy College of   Pharmacy, Maisammaguda, Dhulapally, Secunderabad, Osmania University, Telangana, India.

Corresponding Author E-mail:  srunissa@gmail.com

DOI : http://dx.doi.org/10.13005/bbra/2436

ABSTRACT: The aims of the present study were to extract, purify, characterize and evaluate the in vitro anti -proliferative activity of l-arginase against the cancer cells. L-Arginase is a therapeutic l-arginine depleter, was found to possess excellent antiproliferative activity against various arginine auxotrophic cancer cells. It depletes the levels of l-arginine by converting it into l-ornithine and urea. The marine isolate, Idiomarina sediminium H1695 used in the present study was isolated from coastal areas of Andhra Pradesh, India. The culture was grown and maintained fresh on the nutrient agar slants and preserved under refrigeration. The production of the enzyme was carried out under optimal conditions using submerged fermentation technique. The enzyme was purified to near homogeneity by ammonium sulphate precipitation technique followed by standard chromatographical methods. It was then characterized and evaluated for the anti-proliferative activity against A375-C6 and HCT-116 cells. The results revealed that the enzyme was purified to 29.87 folds after gel filtration chromatography. The purity of the enzyme was confirmed by SDS-PAGE which showed single band of protein with molecular mass 37 k Da. The purified enzyme of Idiomarina sp. exhibited maximum activity at temperature 37°C and pH 7.5. I.C.₅₀ values for A375-C6 and HCT-116 cell lines were found to be 3.47 and 5.765 U/ml respectively. L-Arginase produced by Idiomarina sp. could be a suitable target for arginine auxotrophic cancers. Further animal studies must be carried out to develop the enzyme in the form of chemotherapeutic drug.

KEYWORDS: A375-C6 and HCT-116 cell linesargininosuccinate synthetase;  H1695;  Idiomarina sediminium

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