Biological Activity of Oligomer Chitin Hydrolyzate Produced Using Chitinase Enzymes from SSA2B4.1 (Bacillus cereus SW41) Isolate on Lymphocytes and Cancer Cells Lines.

Abstract

Local chitin waste from crab industries can be used as a source for production of oligomer which has important biological activity. The aims of this research were to evaluate activities of oligomer produced by enzymatic hydrolysis upon proliferation of lymphocytes and cancer cells. the chitinase enzyme was obtained from thermophilic bacterium Bacillus cereus SW41 isolated from South Sulawesi. The medium for producing the enzyme contained 1% colloidal chitin and the enzyme was harvested after four days of incubation, the free cell supernatant were heated at 60oC for 20 minutes. The heat stable protein enzyme was coagulated with 30% saturated ammonium sulphate and purified using hydrophobic interaction chromatography with butyl sepharose gel. The enzyme of 0,005, and 0,0085 IU/mg chitinase were use on 1% chitin colloidal substrate. the reaction products were analyzed and fractionated using HPLC. Cytotoxic assay to determine the lethal concentration 50 (LC50) was done by BSLT method. The effect of oligomers hidrolyzate on lymphocyte proliferative activity and inhibition of cancer cells were conducted by MTT method. The oligomer hidrolyzate proceed by with or without lyophilization at concentration of 62,5 and 125 μg/ml were able to increase lymphocytes proliferation (3-22%). Antiproliferation activity of oligomer chitin hydrolyzate was detected in all tested cancer cell lines with the highest activity occurred in MT2 cell ranging from 17-48% followed by Raji Cell (17-43%), and HeLa cell (5-33%). Based on cell mentioned above, we concluded that oligomer chitin hydrolyzate could be use as an anti cancer cell proliferation and farther study is needed to develop before commercially use.