Two different folding aids including α-cyclodextrin and Mg2+ ions were applied to alkaline phosphatase refolding. The refolding yield depending on concentration of each of the refolding agents reached to 55 and 52% in the presence of 100 mM α-cyclodextrin and/or 5 mM Mg2+ ions, respectively. However, the refolding yield, mediated by combination of α-cyclodextrin and Mg2+ ions, was more than 96%. Replacement of Mg2+ ion with other type of ions which interact with α-cyclodextrin interfere with the function of cyclodextrin resulting in low refolding yields.
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Abbreviations
- ALP:
-
alkaline phosphatase
- pNPP:
-
p-nitrophenyl phosphate
- BSA:
-
bovine serum albumin
- GuHCl:
-
guanidine hydrochloride
- α-CD:
-
α-cyclodextrin
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We would like to thank the research council of the University of Tehran for the financial support of this investigation.
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Khodagholi, F., Yazdanparast, R. Designing a Highly Efficient Refolding System for Alkaline Phosphatase using Combination of Cyclodextrin and Mg2+ Ion. Protein J 27, 1–6 (2008). https://doi.org/10.1007/s10930-006-9021-8
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DOI: https://doi.org/10.1007/s10930-006-9021-8