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

Darwish, Doaa B.; Alrdahe, Salma S.; Al-Awthan, Yahya S.; Elfaki, Imadeldin; Alnour, Tarig M.; Darwish, Ahmed B.; Saad, Entsar A.; Habeb, Salem A.; Youssef, Magdy M.

doi:10.21608/ejchem.2022.148929.6434

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

Document Type : Original Article

Authors

¹ Department of Biology, Faculty of Science, Tabuk University, 71491Tabuk, Saudi Arabia

² Botany Department, Faculty of Science, Mansoura University,35516 Mansoura, Egypt

³ Department of Biology, Faculty of Science, Ibb University, 70270, Ibb, Yemen

⁴ Biochemistry Department, Faculty of Science, Tabuk University, 71491Tabuk, Saudi Arabia

⁵ Medical laboratory technology Department, Faculty of Applied Medical Sciences, Tabuk University, 71491Tabuk, Saudi Arabia

⁶ Zoology Department, Faculty of Science, Suez University, El Salam-1, 43533, Suez, Egypt

⁷ Biochemistry Division, Chemistry Department, Faculty of Science, Damietta University, New Damietta, 34511, Egypt

⁸ Biochemistry Division, Chemistry Department, Faculty of Science, Mansoura University, 35516 Mansoura, Egypt

10.21608/ejchem.2022.148929.6434

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.

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