Abstract

ISOLATION OF THE ANTI-TUMOR L-ASPARAGINASE PRODUCING MICROORGANISMS FROM SWAMPS SOIL IN JAZAN REGION

Dr. Noha Emad Abdel-Razik*, Renad Abdullah Saigh and Khlood Mousa Qashish

ABSTRACT

The objective of this research was to obtain isolates capable of producing high yield of L-asparaginase enzyme and to evaluate the antitumor activity of the purified enzyme against cancer and normal cell lines. Isolation of bacteria was performed by the serial dilution technique of swamp soil samples collected from Jazan, Saudi Arabia, using modified M9 agar plates. Culture filtrates of selected isolates were quantitatively screened for L-asparaginase production using well diffusion and direct nesslerization techniques. The purification of L-asparaginase extracted from isolate was achieved using chilled acetone (-20ºC), followed by gel filtration on Sephadex G-100. The anticancer activity of the purified enzyme against HepGII and WISH cell line was examined by MTT assay. L-asparaginase producer war identified by MicroScan identification system as Bacillus filamentosus. L- asparaginase was extracted, purified and tested in vitro for cytotoxic activity using MMT assay against HepGII and WISH cell line. L-asparaginase from Bacillus filamentosus was neutral to normal epithelial WISH cells. On the other hand, L-asparaginase from the isolate was cytotoxic to HepGII cancer cell line with an IC50 of 75 μg/ml. L-asparaginase extracted showed remarkable anticancer activity. Further studies on hypersensitivity action need to be carried out to recommend the use of L-asparaginase as an alternative to commercially available preparations.

Keywords: L-asparaginase, cytotoxicity, Bacillus filamentosus, HepGII.