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Cloning, expression and purification of Escherichia coli modified phytase
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Malihe Hallaji   , Maryam Parhamfar , Ehsan Raoufi , Hamid Abtahi |
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Abstract: (4223 Views) |
| Introduction: Phytases are the class of phosphatases, which are capable of hydrolyzing phytic acid. Phytases with the phytate degradation are able to reduce or eliminate the harmful effects of phytate. Acidic and thermal stable phytase with high yield and purity by a relatively inexpensive system had extensive application. So, in this study, by modification in enzyme sequence, recombinant phytase production with the shorter length and high expression level was assessed.
Materials and Methods: The phytase gene sequence was obtained from the NCBI database. After bioinformatics studies and doing the noted modification for increasing protein expression, gene proliferation was done by using PCR. E. coli BL21 (DE3) was used to express the protein. Protein purification was performed by Ni-NTA kit and finally, enzyme activity was assessed.
Results: Phytase was successfully expressed and purified. Enzyme activity assay showed a significant activity.
Conclusion: Produced recombinant phytase had high activity in spite of eliminating parts of the enzyme. |
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| Keywords: Molecular Cloning, Phytic Acid, 6-Phytase |
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Full-Text [PDF 521 kb]
(1049 Downloads)
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Type of Study: Research |
Subject:
General
Received: 2017/03/13 | Accepted: 2017/11/15 | Published: 2018/01/2
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