Fig. 4

Enzymatic activities after denaturation with chaotropic agents and renaturation with different proteins. The collapsed enzyme was denatured with the chaotropic agents and was not subjected to renaturation process and serves as negative control. a Percent of α-glucosidase activity measured after denaturation and renaturation assays. α-glucosidase enzyme was denatured with urea (80 mM) and renatured under different treatments, 1 for α-glucosidase; 2 for α-glucosidase + urea; 3 for collapsed α-glucosidase; 4 for Buffer; 5 for BSA; 6 for HtrA; 7 for wild type OppA; 8 for mutant type OppA with double mutation R419G & Y420G; 9 for mutant type OppA with double mutation R41A & D42A; and finally, 10 for mutant type OppA with both double mutations R41A & D42A and R419G & Y420G. The more significant data are shown, HtrA vs. OppAWt with p = 0.104, OppAWt vs. OppA R419G:Y420G with p = 0.472, OppAWt vs. OppA R41A:D42A with p = 0.035, OppAWt vs. OppA R41A:D42A / R419G:Y420 with p = 0.017. b Percentage of LDH activity measured after denaturation and renaturation assays. Lactate-dehydrogenase enzyme was denatured by 1.25 molar guanidine hydrocloride and renatured under different conditions, 1 for LDH; 2 for LDH + GdnHCl; 3 for collapsed LDH; 4 for Buffer; 5 for BSA; 6 for HtrA; 7 for wild type OppA; 8 for mutant type OppA with double mutation R419G & Y420G; 9 for mutant type OppA with double mutation R41A & D42A; and finally, 10 for mutant type OppA with both double mutations R41A & D42A and R419G & Y420G. The more significant data are documented, HtrA vs. OppAWt with p = 0.148, OppAWt vs. OppA R419G:Y420G with p = 0.505, OppAWt vs. OppA R41A:D42A with p = 0.002, OppAWt vs. OppA R41A:D42A / R419G:Y420 with p = 0.0193. The data were analyzed statistically by One-Way ANOVA using Sigma Plot®. The values correspond to the results of three independent repetition assays. Of note, for a detailed interpretation of the result see the main text above