EC Number   |
Reference |
|---|
    3.2.1.1 | acid-denatured enzyme is able to regain the intact structure by neutralization, irreversible denaturation after exposure to an alkaline pH , denaturation under high pressure, reduced enzyme in 8 M urea or 6 M guanidine hydrochloride is completely denatured to a randomly coiled state, Ca2+ has a profound effect on the renaturation process |
393366 |
| 3.2.1.1 | denaturation under high pressure, enzyme denatured by 8 M urea can refold to regain its activity. Rate and extent of reactivation increases in the presence of various other proteins, bovine serum albumin is most effective |
393366 |
| 3.2.1.1 | heat denaturation is irreversible |
393429 |
| 3.2.1.1 | recombinant His6-tagged enzyme lacking the signal peptide from Escherichia coli strain BL21 Star (DE3) inclusion bodies using solubilization of the enzyme protein in denaturing buffer containing 20 mM Tris, pH 8.5, 8 M urea, 100 mM NaCl, 1 mM 2-mercaptoethanol, followed by a on-column refolding method |
750748 |
| 3.2.1.1 | solubilization of recombinant enzyme from inclusion bodies after expression in Escherichia coli without the use of denaturing agents, extraction by resuspending in 50 mmol/l sodium acetate, pH 5.0, containing 20% v/v glycerol followed by stirring at 40°C for 3-4 h, and subsequent centrifugation |
717621 |
| 3.2.1.1 | the enzyme can be unfolded completely by incubating for 4 h with 4 M guanidine HCl or for 12 h with 8 M urea in 0.05 M acetate buffer, pH 4.8, 13.6. mM Ca2+. 0.05 M HEPES buffer, pH 7.0 with 13.6 mM Ca2+ required incubation with 5 M guanidine HCl for 4 h to completely unfold the enzyme. The enzyme refolds on diluting 25 times with HEPES buffer containing 15 mM dithiothreitol, resulting in the recovery of ca. 85% of the activity. The refolded enzyme regains secondary and tertiary structure similar to the native enzyme. Refolding experiments with the acetate buffer do not yield active enzyme. alpha-Amlyase is more resistant to unfolding at pH 7.0 in comparison to unfolding at pH 4.8. The enzyme shows significant unfolding at 27°C, at urea concentrations below 9 M, pH 7.0 |
691135 |
