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. 2019 May 30;85(12):e00275-19. doi: 10.1128/AEM.00275-19

FIG 5.

FIG 5

Purification and characterization of the His6-DMFase protein. (A) SDS-PAGE analysis of His6-DMFase. Lane 1, sonicated E. coli BL21(DE3)-pETDuet-1; lane 2, sonicated E. coli BL21(DE3)-pETDuet-1-dmfA1A2; lane 3, E. coli BL21(DE3)-pETDuet-dmfA1A2 pellet; lane 4, E. coli BL21(DE3)-pETDuet-dmfA1A2 supernatant; lane 5, purified His6-DMFase; lane M, molecular size markers. (B) Effect of pH on His6-DMFase activity. (C) Effect of temperature on His6-DMFase activity. (D) Effect of metal ions (1.0 mM) on His6-DMFase activity. (E) In vitro assays of enzyme activity using an alkyl-amine-specific color reaction, with DMF used as the substrate (a) or no DMF added (b). (F) Specific enzyme activities for His6-DMFase incubated overnight at different temperatures. (G) Circular dichroism spectra of DMFase at increasing temperatures from 25°C to 75°C. (H) Kinetic analysis of His6-DMFase (fitted to a Michaelis-Menten curve).