Purification, Molecular Characterization, and Anticancer Activities of L- Asparaginase extracted from Staphylococcus aureus
Darwish, D., Alrdahe, S., Al-Awthan, Y., Elfaki, I., Alnour, T., Darwish, A., Saad, E., Habeb, S., Youssef, M. (2023). Purification, Molecular Characterization, and Anticancer Activities of L- Asparaginase extracted from Staphylococcus aureus. EKB Journal Management System, 66(5), 245-259. doi: 10.21608/ejchem.2022.148929.6434
Doaa B. Darwish; Salma S. Alrdahe; Yahya S. Al-Awthan; Imadeldin Elfaki; Tarig M. Alnour; Ahmed B. Darwish; Entsar A. Saad; Salem A. Habeb; Magdy M. Youssef. "Purification, Molecular Characterization, and Anticancer Activities of L- Asparaginase extracted from Staphylococcus aureus". EKB Journal Management System, 66, 5, 2023, 245-259. doi: 10.21608/ejchem.2022.148929.6434
Darwish, D., Alrdahe, S., Al-Awthan, Y., Elfaki, I., Alnour, T., Darwish, A., Saad, E., Habeb, S., Youssef, M. (2023). 'Purification, Molecular Characterization, and Anticancer Activities of L- Asparaginase extracted from Staphylococcus aureus', EKB Journal Management System, 66(5), pp. 245-259. doi: 10.21608/ejchem.2022.148929.6434
Darwish, D., Alrdahe, S., Al-Awthan, Y., Elfaki, I., Alnour, T., Darwish, A., Saad, E., Habeb, S., Youssef, M. Purification, Molecular Characterization, and Anticancer Activities of L- Asparaginase extracted from Staphylococcus aureus. EKB Journal Management System, 2023; 66(5): 245-259. doi: 10.21608/ejchem.2022.148929.6434

Purification, Molecular Characterization, and Anticancer Activities of L- Asparaginase extracted from Staphylococcus aureus

Egyptian Journal of Chemistry
Volume 66, Issue 5, May 2023, Page 245-259  XML PDF (1.51 MB)
Document Type: Original Article
DOI: 10.21608/ejchem.2022.148929.6434
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Authors
Doaa B. Darwish1, 2; Salma S. Alrdahe1; Yahya S. Al-Awthan1, 3; Imadeldin Elfaki4; Tarig M. Alnourorcid 5; Ahmed B. Darwish6; Entsar A. Saad7; Salem A. Habeb4; Magdy M. Youssef email orcid 8
1Department of Biology, Faculty of Science, Tabuk University, 71491Tabuk, Saudi Arabia
2Botany Department, Faculty of Science, Mansoura University,35516 Mansoura, Egypt
3Department of Biology, Faculty of Science, Ibb University, 70270, Ibb, Yemen
4Biochemistry Department, Faculty of Science, Tabuk University, 71491Tabuk, Saudi Arabia
5Medical laboratory technology Department, Faculty of Applied Medical Sciences, Tabuk University, 71491Tabuk, Saudi Arabia
6Zoology Department, Faculty of Science, Suez University, El Salam-1, 43533, Suez, Egypt
7Biochemistry Division, Chemistry Department, Faculty of Science, Damietta University, New Damietta, 34511, Egypt
8Biochemistry Division, Chemistry Department, Faculty of Science, Mansoura University, 35516 Mansoura, Egypt
Abstract
L- asparaginases catalyze the conversion of L- asparagine to L- aspartate and ammonia. The current study focus on the cloning and expression of the L-asparaginase from Staphylococcus aureus into Escherichia coli strain BL21(DE3)pLysS. L- asparaginase enzyme was purified to homogeneity by glutathione sepharose 4B column chromatography. The enzyme was purified 117.6 times and showed a final specific activity of 1680.4 IU/mg protein with a yield of 67.7%. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified enzyme showed that it was a single peptide chain with Mr 35 kDa. The enzyme was immobilized on Ca alginate beads. The immobilized enzyme retains most of its activity (78%) and reveals high stability at 4 °C. The enzymatic and structural properties of free recombinant and immobilized L- asparaginase were studied. The free enzyme showed maximum activity at pH 8.0 when incubated at 45 °C for 30 min. The immobilized enzyme displayed maximum activity at pH 8.5 after 30 minutes of incubation at 50 °C. The amino acid composition of the purified enzyme was documented. This approach offers the possibility of generating Staphylococcus aureus L- asparaginase with high efficiency that can be used to treat leukemia.
Keywords
Staphylococcus aureus - L Asparaginase – Overexpression- Purification- Immobilization; Optimization
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