Purification and properties of a phytate-degrading enzyme produced by Enterobacter sakazakii ASUIA279.
An extracellular phytate-degrading enzyme produced by Enterobacter sakazakii ASUIA279 was purified to homogeneity using FPLC anion exchange chromatography and gel filtration. The enzyme was purified about 66-fold with a recovery of 27%. Its molecular mass was estimated to be 43 kDa by SDS-PAGE. The...
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| Format: | Article |
| Language: | English English |
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Federal University of Tocantins
2012
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| Online Access: | http://psasir.upm.edu.my/id/eprint/24200/ http://psasir.upm.edu.my/id/eprint/24200/1/Purification%20and%20properties%20of%20a%20phytate.pdf |
| _version_ | 1848844969655140352 |
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| author | Farouk, Abd-ElAziem Greiner, Ralf Meor Hussin, Anis Shobirin |
| author_facet | Farouk, Abd-ElAziem Greiner, Ralf Meor Hussin, Anis Shobirin |
| author_sort | Farouk, Abd-ElAziem |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | An extracellular phytate-degrading enzyme produced by Enterobacter sakazakii ASUIA279 was purified to homogeneity using FPLC anion exchange chromatography and gel filtration. The enzyme was purified about 66-fold with a recovery of 27%. Its molecular mass was estimated to be 43 kDa by SDS-PAGE. The Michaelis constant (KM) and turnover number (kcat ) for sodium phytate at pH 5.0 and 50°C were calculated from the Lineweaver-Burk plot to be 760 µM and 4.14s-1, respectively. The enzyme showed narrow substrate specificity and not phytate, but GTP was dephosphorylated with the highest relative rate of hydrolysis. However, according to the kcat/KM values, phytate was concluded to be the in vivo substrate of the enzyme. Optimal activity was determined at pH 4.5 and 45-55°C. The enzyme was strongly inhibited by Fe3+, Cu2+, Zn2+, molybdate, vanadate, fluoride and phosphate (1 mM). |
| first_indexed | 2025-11-15T08:39:22Z |
| format | Article |
| id | upm-24200 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English English |
| last_indexed | 2025-11-15T08:39:22Z |
| publishDate | 2012 |
| publisher | Federal University of Tocantins |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-242002015-09-18T08:04:52Z http://psasir.upm.edu.my/id/eprint/24200/ Purification and properties of a phytate-degrading enzyme produced by Enterobacter sakazakii ASUIA279. Farouk, Abd-ElAziem Greiner, Ralf Meor Hussin, Anis Shobirin An extracellular phytate-degrading enzyme produced by Enterobacter sakazakii ASUIA279 was purified to homogeneity using FPLC anion exchange chromatography and gel filtration. The enzyme was purified about 66-fold with a recovery of 27%. Its molecular mass was estimated to be 43 kDa by SDS-PAGE. The Michaelis constant (KM) and turnover number (kcat ) for sodium phytate at pH 5.0 and 50°C were calculated from the Lineweaver-Burk plot to be 760 µM and 4.14s-1, respectively. The enzyme showed narrow substrate specificity and not phytate, but GTP was dephosphorylated with the highest relative rate of hydrolysis. However, according to the kcat/KM values, phytate was concluded to be the in vivo substrate of the enzyme. Optimal activity was determined at pH 4.5 and 45-55°C. The enzyme was strongly inhibited by Fe3+, Cu2+, Zn2+, molybdate, vanadate, fluoride and phosphate (1 mM). Federal University of Tocantins 2012 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/24200/1/Purification%20and%20properties%20of%20a%20phytate.pdf Farouk, Abd-ElAziem and Greiner, Ralf and Meor Hussin, Anis Shobirin (2012) Purification and properties of a phytate-degrading enzyme produced by Enterobacter sakazakii ASUIA279. Journal of Biotechnology and Biodiversity , 3 (1). pp. 1-9. ISSN 2179-4804 http://www.uft.edu.br/ English |
| spellingShingle | Farouk, Abd-ElAziem Greiner, Ralf Meor Hussin, Anis Shobirin Purification and properties of a phytate-degrading enzyme produced by Enterobacter sakazakii ASUIA279. |
| title | Purification and properties of a phytate-degrading enzyme produced by Enterobacter sakazakii ASUIA279. |
| title_full | Purification and properties of a phytate-degrading enzyme produced by Enterobacter sakazakii ASUIA279. |
| title_fullStr | Purification and properties of a phytate-degrading enzyme produced by Enterobacter sakazakii ASUIA279. |
| title_full_unstemmed | Purification and properties of a phytate-degrading enzyme produced by Enterobacter sakazakii ASUIA279. |
| title_short | Purification and properties of a phytate-degrading enzyme produced by Enterobacter sakazakii ASUIA279. |
| title_sort | purification and properties of a phytate-degrading enzyme produced by enterobacter sakazakii asuia279. |
| url | http://psasir.upm.edu.my/id/eprint/24200/ http://psasir.upm.edu.my/id/eprint/24200/ http://psasir.upm.edu.my/id/eprint/24200/1/Purification%20and%20properties%20of%20a%20phytate.pdf |