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Abstract

Misfolding poses a serious problem in the biotechnological field in obtaining the active protein from inclusion bodies. Here we show that high temperature increases the refolding yield of reduced lyosyzme by a simple dilution method. The refolding yields at 98 degrees C were three times higher than those at 20 degrees C in the solutions tested, which is related to the fact that the thermally unfolded state of lysozyme is a more productive form for folding than the denaturant-induced fully unfolded state. The thermal-assisted refolding could be used for various reduced and denatured proteins as a result of its simplicity and low cost.

Mechanistic insights into protein precipitation by alcohol

“…Reduced and denatured lysozyme was prepared as previously reported [22]. Briefly, lysozyme was dissolved at 20 mg/ml in water containing 6 M Gdn • HCl, 1 mM EDTA and 40 mM DTT.…”

Effects of alcohol on the solubility and structure of native and disulfide-modified bovine serum albumin

“…Reduced BSA was prepared as reported previously [9,13]. Briefly, BSA was dissolved at 20 mg/mL in water containing 6 M GdnHCl, 1 mM EDTA, and 40 mM DTT.…”

Alumina nanoparticle-assisted enzyme refolding: A versatile methodology for proteins renaturation

“…In water, colloidal or interfacial assemblies with surfactants or nanoparticles provide conditions to control protein stability and folding/unfolding rearrangements and renaturation 3 . Several methods based on interface interactions for renaturation of proteins are available, which in general can be divided into three main groups: (I) methods to remove the denaturants from the protein solution 4–6 ; (II) methods involving the control of the physical conditions used during the refolding process 7, 8 ; and (III) methods in which refolding agents are added during the renaturing processes 9–11 . A major drawback of these methods has been the high specificity to a particular protein which requires tailoring of conditions in each case; from that point of view, a versatile chemical general tool, applicable for any target protein is yet to be developed, and is highly needed.…”