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Journal of Biological Sciences

Year: 2020 | Volume: 20 | Issue: 1 | Page No.: 48-55
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Authors

Mohamed Abdel-Raouf

Mohamed Abdel-Raouf

Hisham M. Aldeweik

Ehab Y. Elbendary

Ehab Y. Elbendary

Keywords

Research Article

Impact of Physical and Chemical Factors on L-asparaginase Enzyme Purified from Soil Bacteria

Mohamed Abdel-Raouf Mohamed Abdel-Raouf's LiveDNA, Hisham M. Aldeweik and Ehab Y. Elbendary Ehab Y. Elbendary's LiveDNA
Background and Objective: L- Asparaginase is one of the amidohydrolases enzymes which catalyses the deamination of asparaginine to aspartic acid. Our study aimed to produce and purify L-asparaginase enzyme from soil bacteria and investigate the physical and chemical factors that effect on the amount of produced L-asparaginase enzymes. Materials and Methods: Fifty soil samples were collected from soil surface, the soil suspension was then inoculated on selective nutrient agar medium, Isolated bacterial colonies were identified by biochemical tests to select the most active bacterial strains for production of L-asparaginase enzyme. The L-asparaginase enzyme was purified by Sephadex G-100 column. The effects of temperature, pH, different nutritional factors were tested on the amount of produced L-asparaginase enzyme. Results: Erwinia carotovora has the highest L-asparaginase enzyme activity with 14 mm of intense pink zone, followed by E. coli (13 mm) while Streptococcus pyogenes has (12 mm). Optimal conditions for L-asparaginase enzyme production from Erwinia carotovora were pH 7.0 at 37°C. The best carbon source was glucose with activity (2.7) and best concentration of glucose was 10 g L–1. The best nitrogen source was L-asparagine by concentration of 5 g L–1 which gave activity (4.7). Finally this study indicated that the maximum L-asparaginase activity was obtained after incubation for 10 min at 37°C, pH 8.0 with adding 1.0 mL of magnesium chloride (MgCl2) as activator. Conclusion: Our study concluded that the soil can be used for obtaining L-asparaginase produced bacteria and the Erwinia carotovora is consider as the most active bacteria for L-asparaginase production.
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