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Overexpression, biochemical characterization, and anticancer activates of L-asparaginase from Bacillus subtilis
- Source :
- International journal of health sciences. :1731-1752
- Publication Year :
- 2022
- Publisher :
- Universidad Tecnica de Manabi, 2022.
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Abstract
- L-asparaginases convert L-asparagine into L-aspartate and ammonia. The L-asparaginase from Bacillus subtilis was cloned and expressed in the E. coli strain BL21(DE3)pLysS in the current study. Using glutathione sepharose 4B column chromatography, the L-asparaginase enzyme was uniformly purified 173.34 times, with a final specific activity of 1769.13 IU/mg protein and a yield of 56.14%. The isolated enzyme was identified as a 36 kDa polypeptide chain by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The immobilized enzyme was placed on top of the Ca alginate beads. The immobilized enzyme is quite stable and retains the majority of its activity at 4 °C (74 percent). The enzymatic and structural characteristics of free and immobilized recombinant L-asparaginase were studied. The activity of the free enzyme peaked after 30 min of incubation at pH 8.0 and 45 °C. After 30 minutes at 50 °C, the immobilized enzyme showed its peak activity at a pH of 8.5. The refined enzyme's amino acid makeup was identified. An enzyme that heals leukemia, Bacillus subtilis L-asparaginase, can be successfully mass-produced using this technique.
- Subjects :
- General Nursing
Education
Subjects
Details
- ISSN :
- 2550696X and 25506978
- Database :
- OpenAIRE
- Journal :
- International journal of health sciences
- Accession number :
- edsair.doi...........5d91e672c29ba1e04ecdbccec2555dde
- Full Text :
- https://doi.org/10.53730/ijhs.v6ns8.11477