Characterization and Antitumor Activity Study for a Therapeutically Important L-glutaminase Purified from Staphylococcus Aureus

1Mustafa Gh. AL-Juha ishy. Asmaa A. Hussein

166 Views
56 Downloads
Abstract:

L-glutaminase enzyme produced under optimum conditions in a previous study was purified using precipitation with 80% saturation of sulfate ammonium, ion exchange chromatography on column DEAE-cellulose, and filtration gel chromatography during Sephadex G-200 column, the activity specific of the purified enzyme after last step was raised to 1000 U/mg for 38.4 folds of purification and 69% recovery enzyme. MWut. of purified lglutaminase was estimation through S.D.S-P.A.G.E to be 35 KDa, It was gave highest activity at pH 8.0 and stability at pH 7.5, the enzyme was active at 37 oC and stable at a range of temperatures from 20oC to 37 oC. Also lglutaminase activity was inhibited to (62%) in the presence of CaCl2. L-glutaminase was tested for in-vitro cytotoxic activity using MTT assay against colorectal adenocarcinoma (LS 174T) cell line which was inhibited with an IC50 of 37.19 IU\ml however, it didn’t show a important affect on normal cell.

Keywords:

L-Glutaminase, MTT Assay, LS 174T Cell Line.

Paper Details
Month4
Year2020
Volume24
IssueIssue 5
Pages3029-3037