PARTIAL PURIFICATION AND CYTOTOXIC ACTIVITY OFL-ASPARAGINASE ISOLATED FROM Escherichia coli

Ayat   Abbas; Majeed Sabbah; Omer   Kathum

doi:10.24996/ijs.2010.51.2.%g

Authors

Ayat Abbas Biotechnology Research Center, University of Al Nahrain. Baghdad-Iraq.
Majeed Sabbah Biotechnology Research Center, University of Al Nahrain. Baghdad-Iraq.
Omer Kathum Biotechnology Research Center, University of Al Nahrain. Baghdad-Iraq.

DOI:

https://doi.org/10.24996/ijs.2010.51.2.%25g

Keywords:

PARTIAL, CYTOTOXIC

Abstract

L-Asparaginase (E.C.3.5.1.1) is an important natural product that possesses a
broad spectrum of antitumor activity. In the present study, L-asparaginase partially
purified from local isolate of Escherichia coli that were grown aerobically for four
hours and anaerobically for 18hrs on M9 medium contain L-asparagine as sole
nitrogen source. Extraction of the enzyme was done by sonication. Qualitative and
quantitative assays for L-Asparaginase production were determined using
colorimetric and nesslerization methods respectively. The purification steps involve
dialysis of crude extract and DEAE-Cellulose Ion exchange chromatography. The
enzyme was purified 3.39-folds and showed a final specific activity of 0.18 U/mg
with a 41% yield. The crude extract and DEAE-Cellulose fractions showed slight
growth inhibition against RD cell line (human rhambdomyo sarcoma).