Purification and characterization of L-asparaginase produced from Bacillus sp.
The objective of this study was to isolate and identify the asparaginase-producing bacteria, then purify and characterize the enzyme in order to investigate their properties in the future. Fifteen local bacterial isolates were isolated from various sites in the city of Baghdad, identified by conven...
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University of Baghdad, College of Science for Women
2023-12-01
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| Series: | مجلة بغداد للعلوم |
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| Online Access: | https://bsj.uobaghdad.edu.iq/index.php/BSJ/article/view/9231 |
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| author | Sahar I. H. Zaid A. H. Milad A. |
| author_facet | Sahar I. H. Zaid A. H. Milad A. |
| author_sort | Sahar I. H. |
| collection | DOAJ |
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The objective of this study was to isolate and identify the asparaginase-producing bacteria, then purify and characterize the enzyme in order to investigate their properties in the future. Fifteen local bacterial isolates were isolated from various sites in the city of Baghdad, identified by conventional morphological and biochemical procedures, and confirmed using vitek 2 methods, and submitted to primary screening processes for asparaginase production. For secondary screening, eight isolates with the greatest yellow zone ability on a specific solid medium were chosen. Bacillus sp. was reported to have the highest enzyme production (7.5 U/mg proteins). After 24 hours of incubation, submerged fermentation yielded optimal conditions for the production of L-asparaginase (L-ASNase) by the chosen isolate, with medium (2) serving as the optimal medium for production and fructose serving as the optimal source of carbon. In pH 6 at 40°C, Sephadex G-150 gel filtration chromatography was used to purify the enzyme. The final purification folds were increased by 2.5 times, resulting in an enzyme yield of 93.7%. It also showed the highest purified enzyme activity and stability was at 37°C. Also it revealed the highest activity and stability at pH 7.0 and pH 8.0 respectively. Enzyme lost activity when exposed to several metallic ions at concentrations of 1, 5, and 10 mM.
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| format | Article |
| id | doaj-art-e39d4724b663446ca949c2ebeda3425c |
| institution | DOAJ |
| issn | 2078-8665 2411-7986 |
| language | English |
| publishDate | 2023-12-01 |
| publisher | University of Baghdad, College of Science for Women |
| record_format | Article |
| series | مجلة بغداد للعلوم |
| spelling | doaj-art-e39d4724b663446ca949c2ebeda3425c2025-08-20T03:19:07ZengUniversity of Baghdad, College of Science for Womenمجلة بغداد للعلوم2078-86652411-79862023-12-01206(Suppl.)10.21123/bsj.2023.9231Purification and characterization of L-asparaginase produced from Bacillus sp.Sahar I. H.0Zaid A. H.1Milad A.2Biotechnology Department, College of Science, University of Baghdad, Baghdad, Iraq.Biotechnology Department, College of Science, University of Baghdad, Baghdad, Iraq.Iraqi Ministry of Health, Baghdad, Iraq. The objective of this study was to isolate and identify the asparaginase-producing bacteria, then purify and characterize the enzyme in order to investigate their properties in the future. Fifteen local bacterial isolates were isolated from various sites in the city of Baghdad, identified by conventional morphological and biochemical procedures, and confirmed using vitek 2 methods, and submitted to primary screening processes for asparaginase production. For secondary screening, eight isolates with the greatest yellow zone ability on a specific solid medium were chosen. Bacillus sp. was reported to have the highest enzyme production (7.5 U/mg proteins). After 24 hours of incubation, submerged fermentation yielded optimal conditions for the production of L-asparaginase (L-ASNase) by the chosen isolate, with medium (2) serving as the optimal medium for production and fructose serving as the optimal source of carbon. In pH 6 at 40°C, Sephadex G-150 gel filtration chromatography was used to purify the enzyme. The final purification folds were increased by 2.5 times, resulting in an enzyme yield of 93.7%. It also showed the highest purified enzyme activity and stability was at 37°C. Also it revealed the highest activity and stability at pH 7.0 and pH 8.0 respectively. Enzyme lost activity when exposed to several metallic ions at concentrations of 1, 5, and 10 mM. https://bsj.uobaghdad.edu.iq/index.php/BSJ/article/view/9231Asparaginase, Bacteria, Optimum conditions, Purification, Characterization |
| spellingShingle | Sahar I. H. Zaid A. H. Milad A. Purification and characterization of L-asparaginase produced from Bacillus sp. مجلة بغداد للعلوم Asparaginase, Bacteria, Optimum conditions, Purification, Characterization |
| title | Purification and characterization of L-asparaginase produced from Bacillus sp. |
| title_full | Purification and characterization of L-asparaginase produced from Bacillus sp. |
| title_fullStr | Purification and characterization of L-asparaginase produced from Bacillus sp. |
| title_full_unstemmed | Purification and characterization of L-asparaginase produced from Bacillus sp. |
| title_short | Purification and characterization of L-asparaginase produced from Bacillus sp. |
| title_sort | purification and characterization of l asparaginase produced from bacillus sp |
| topic | Asparaginase, Bacteria, Optimum conditions, Purification, Characterization |
| url | https://bsj.uobaghdad.edu.iq/index.php/BSJ/article/view/9231 |
| work_keys_str_mv | AT saharih purificationandcharacterizationoflasparaginaseproducedfrombacillussp AT zaidah purificationandcharacterizationoflasparaginaseproducedfrombacillussp AT milada purificationandcharacterizationoflasparaginaseproducedfrombacillussp |