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Purification and characterization of deoxyribonuclease from small intestine of camel Camelus dromedarius
The chromatography of deoxyribonuclease (DNase) from small intestine of camel Camelus dromedarius by DEAE-Sepharose separated three isoforms DNase 1, DNase 2 and DNase 3. The DNase 3 was purified to homogeneity by chromatography on Sephacryl S-200. The molecular weight of DNase 3 was 30 kDa using ge...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Academy of Scientific Research and Technology, Egypt
2017
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6296583/ https://www.ncbi.nlm.nih.gov/pubmed/30647687 http://dx.doi.org/10.1016/j.jgeb.2017.06.008 |
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author | Abdel-Gany, Somia S. El-Badry, Mohamed O. Fahmy, Afaf S. Mohamed, Saleh A. |
author_facet | Abdel-Gany, Somia S. El-Badry, Mohamed O. Fahmy, Afaf S. Mohamed, Saleh A. |
author_sort | Abdel-Gany, Somia S. |
collection | PubMed |
description | The chromatography of deoxyribonuclease (DNase) from small intestine of camel Camelus dromedarius by DEAE-Sepharose separated three isoforms DNase 1, DNase 2 and DNase 3. The DNase 3 was purified to homogeneity by chromatography on Sephacryl S-200. The molecular weight of DNase 3 was 30 kDa using gel filtration and SDS-PAGE. The pH optimum of DNase 3 was reported at 7.0 using Tris-HCl buffer. The temperature optimum of DNase 3 was found to be 50 °C. The enzyme was stable up to 50 °C for one h incubation. The Km value was 28.5 µg DNA, where this low value indicated the high affinity of enzyme toward DNA as substrate. No activity of DNase 3 was determined in the absence of metal cations. Mg(2+) and Ca(2+) caused significant enhancement in the enzyme activity by 90 and 75%, respectively. The mixture of Mg(2+) and Ca(2+) caused 100% of enzyme activity. Ni(2+), Co(2+), Ba(2+), Zn(2+) and Cd(2+) showed very strong inhibitory effect on enzyme activity. In conclusion, the characterization of DNase 3 indicated that the enzyme is considered as a member of DNase I family. The low Km value of the DNA suggested that the high digestion of DNA of camel forage by small intestine DNase 3. |
format | Online Article Text |
id | pubmed-6296583 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Academy of Scientific Research and Technology, Egypt |
record_format | MEDLINE/PubMed |
spelling | pubmed-62965832019-01-15 Purification and characterization of deoxyribonuclease from small intestine of camel Camelus dromedarius Abdel-Gany, Somia S. El-Badry, Mohamed O. Fahmy, Afaf S. Mohamed, Saleh A. J Genet Eng Biotechnol Animal Biotechnology The chromatography of deoxyribonuclease (DNase) from small intestine of camel Camelus dromedarius by DEAE-Sepharose separated three isoforms DNase 1, DNase 2 and DNase 3. The DNase 3 was purified to homogeneity by chromatography on Sephacryl S-200. The molecular weight of DNase 3 was 30 kDa using gel filtration and SDS-PAGE. The pH optimum of DNase 3 was reported at 7.0 using Tris-HCl buffer. The temperature optimum of DNase 3 was found to be 50 °C. The enzyme was stable up to 50 °C for one h incubation. The Km value was 28.5 µg DNA, where this low value indicated the high affinity of enzyme toward DNA as substrate. No activity of DNase 3 was determined in the absence of metal cations. Mg(2+) and Ca(2+) caused significant enhancement in the enzyme activity by 90 and 75%, respectively. The mixture of Mg(2+) and Ca(2+) caused 100% of enzyme activity. Ni(2+), Co(2+), Ba(2+), Zn(2+) and Cd(2+) showed very strong inhibitory effect on enzyme activity. In conclusion, the characterization of DNase 3 indicated that the enzyme is considered as a member of DNase I family. The low Km value of the DNA suggested that the high digestion of DNA of camel forage by small intestine DNase 3. Academy of Scientific Research and Technology, Egypt 2017-12 2017-06-28 /pmc/articles/PMC6296583/ /pubmed/30647687 http://dx.doi.org/10.1016/j.jgeb.2017.06.008 Text en © 2017 Production and hosting by Elsevier B.V. on behalf of Academy of Scientific Research & Technology. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Animal Biotechnology Abdel-Gany, Somia S. El-Badry, Mohamed O. Fahmy, Afaf S. Mohamed, Saleh A. Purification and characterization of deoxyribonuclease from small intestine of camel Camelus dromedarius |
title | Purification and characterization of deoxyribonuclease from small intestine of camel Camelus dromedarius |
title_full | Purification and characterization of deoxyribonuclease from small intestine of camel Camelus dromedarius |
title_fullStr | Purification and characterization of deoxyribonuclease from small intestine of camel Camelus dromedarius |
title_full_unstemmed | Purification and characterization of deoxyribonuclease from small intestine of camel Camelus dromedarius |
title_short | Purification and characterization of deoxyribonuclease from small intestine of camel Camelus dromedarius |
title_sort | purification and characterization of deoxyribonuclease from small intestine of camel camelus dromedarius |
topic | Animal Biotechnology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6296583/ https://www.ncbi.nlm.nih.gov/pubmed/30647687 http://dx.doi.org/10.1016/j.jgeb.2017.06.008 |
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