Designing a highly efficient chemical chaperone system using chitosan-coated alginate.
In the present work we prepared chitosan-coated alginate beads, to use as a chemical chaperone based on the electrostatic interaction between the carboxylate groups of alginate and the ammonium groups of chitosan. This procedure was an attempt for designing a highly efficient chemical chaperone to improve protein stability and refolding. Based on enzyme recovered activity, turbidity, far-UV CD and fluorescence data, alkaline phosphatase can be stabilized and refolded to a higher degree in the presence of alginate capsules compared with unassisted form and was further improved by including chitosan. Finally the maximum yield was obtained when the refolding process was achieved under a well worked out temperature program: incubation of the captured-enzyme for 20 min at 4 degrees C followed by overnight incubation at 22 degrees C, which showed that aggregation is a major limitation to refolding.
Khodagholi
Fariba
F
Neuroscience Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran. khodagholi@sbmu.ac.ir
Farahmand
Shahrzad
S
Tusi
Solaleh Khoramian
SK
D000464:Alginates / Q000737:chemistry*; D000469:Alkaline Phosphatase / Q000737:chemistry / Q000378:metabolism; D000704:Analysis of Variance; D048271:Chitosan / Q000737:chemistry / Q000378:metabolism; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D008863:Microspheres; D018832:Molecular Chaperones / Q000737:chemistry / Q000378:metabolism; D017510:Protein Folding; D055550:Protein Stability; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013696:Temperature
D000464:Alginates; D006603:Hexuronic Acids; D018832:Molecular Chaperones; D011994:Recombinant Proteins; D020723:Glucuronic Acid; D048271:Chitosan; D000469:Alkaline Phosphatase