Anticancer Activity of Cytosine Deaminase Enzyme Purified From Local Saccharomyces cerevisia Bread Yeast

Document Type : Original Article

Authors

1 Biotechnology Research Center-Al-Nahrain university

2 Collage of Education for Pure Sciences\ University Tikrit, Salah al-din , Iraq

3 Biology,collage of Education for pure Sciences,University of Tikrit, Salah l-din.Iraq

Abstract

The study aimed to extract and purify the enzyme cytosine deaminase from locally manufactured bread yeast Saccharomyces cerevisia. To achieve the goal of the study, bread yeast was made locally and transported to the laboratory for enzyme extraction process using toluene as organic solvent for yeast wall rapturing. Crude enzyme extracted with cold distilled water. Steps for enzyme purification starting by precipitation step with ammonium sulfate 60% saturation, then ion exchange purification method, ended with gel filtration purification proses had been applied. In each step, the supernatant volume, enzyme activity, specific enzyme activity, protein concentration, and percentage of enzyme yield had been recorded. The cytotoxic effect of different enzyme concentrations in all purification steps toward the lung cancer cell line A549 and human breast cancer line MDA cell line beside normal cell line REF cells was investigated. The specific activity of the crude extract of locally produced bread yeast reached 13.205 units/mg protein, and for ammonium sulfate enzyme precipitation the specific activity was amounted 17.68 units/mg protein, for ion exchange was 216.66 units/mg protein, while for purification with gel filtration the specific activity reached 571.428 unit/mg protein. The cytotoxic effect of the extracted enzyme in all steps; crud, ion-exchange, and gel filtration purified enzyme on the lungA549 and breast AMD cancer lines beside the REF normal cells were should a range of toxic effect at different enzyme concentration. The toxicity was reduced till diminishing as the enzyme was more purified.

Keywords

Journal of Bioscience and Applied Research

Articles in Press, Accepted Manuscript
Available Online from 06 March 2024

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