Designing a highly efficient refolding system for alkaline phosphatase using combination of cyclodextrin and Mg2+ ion.
Two different folding aids including alpha-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 alpha-cyclodextrin and/or 5 mM Mg2+ ions, respectively. However, the refolding yield, mediated by combination of alpha-cyclodextrin and Mg2+ ions, was more than 96%. Replacement of Mg2+ ion with other type of ions which interact with alpha-cyclodextrin interfere with the function of cyclodextrin resulting in low refolding yields.
Khodagholi
Fariba
F
Institute of Biochemistry and Biophysics, The University of Tehran, P.O. Box 13145-1384, Tehran, Iran.
Yazdanparast
Razieh
R
D000469:Alkaline Phosphatase / Q000737:chemistry*; D002942:Circular Dichroism; D004789:Enzyme Activation; D008274:Magnesium / Q000737:chemistry*; D011489:Protein Denaturation; D017510:Protein Folding; D047391:alpha-Cyclodextrins / Q000737:chemistry*
D047391:alpha-Cyclodextrins; D000469:Alkaline Phosphatase; D008274:Magnesium; C032613:alpha-cyclodextrin