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Biochemical Study of Alkaline Phosphatase inStaphylococcus aureus | ||
| Rafidain Journal of Science | ||
| Article 39, Volume 27, Issue 4, 2018, Pages 271-286 PDF (608.25 K) | ||
| DOI: 10.33899/rjs.2018.159326 | ||
| Authors | ||
| Najwa I. Khaleel; Ibtihal E. Kanaan | ||
| Abstract | ||
| The present research aimed isolating and purifying Alkaline phosphatase enzyme from crude protein extract (Lysate supernatant) of Staphylococcus aureus by using different biotechnologies. To proceed, the following steps were achieved:Firstly, The verification of the existence of enzyme in bacteria, the bacteria were diagnosed by using the API 20 stripe that consists of (20) items. It had been detected that the experimented bacterium was Staphylococcus aureus, the enzyme was isolated from this bacterium to ensure its availability in it within the logarithmic phase and this was done through growing it for 18 hours in a suitable growth medium. It had been detected that the enzyme was intracellular because of the occurrence of enzyme activity in the lysate supernatant without occurring it in the cell free culture supernatant. Secondly, Enzyme purification, the enzyme had been purified through three stages: precipitation of protein by ammonium sulphate, dialysis and finally, the protein extract was passed through column chromatography by using Sephadex G-100 gel, the estimated enzyme activity after this step was 16.2 in comparison with its activity before the purification processes (crude protein extract). The approximate molecular weight of alkaline phosphatase was estimated by using gelatinous filtration technique and it was 51.000 Dalton. Thirdly, Measuring of the enzyme activity in the experimented animals, the results showed an increase of enzyme activity in the blood serum of mice injected with pathological bacteria in comparison with its activity in the blood serum of healthy mice, i.e. the intact ones. ). | ||
| Keywords | ||
| alkaline phosphatase; Staphylococcus aureus; molecular weight | ||
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