Skip to main content
Figure 3 | Microbial Cell Factories

Figure 3

From: A comparative summary of expression systems for the recombinant production of galactose oxidase

Figure 3

SDS-PAGE analyses of recombinantly produced GalOx . A. Fractions collected from the IMAC purification of GalOxM1 expressed intracellularly in E. coli at 25°C (incubated with 0.5 mM CuSO4 before loading on the gel); lane 1, molecular mass standard; lane 2, crude extract; lane 3, flow-through; lane 4-7, different fractions of GalOxM1 eluted with a linear gradient of imidazole (20 mM to 1 M imidazole within 5 column volumes). B. Crude extracts obtained after 120 h cultivation of P. pastoris expressing the wtgalox or the variant M1galox either with or without an N-terminal His10-tag at 25°C (intracellular production); lane 1, molecular mass standard; lane 2, crude extract of His10-wtGalOx; lane 3, crude extract of His10-M1GalOx; lane 4, crude extract of wtGalOx; lane 5, crude extract of M1GalOx. C. Aliquots of the supernatant obtained at different time points during the cultivation of P. pastoris expressing the wtgalox gene from F. graminearum without the N-terminal His10-tag extracellularly at 25°C; lane 1, molecular mass standard; lane 2-5, 48 h; 72 h; 96 h (1:10 dilution); 120 h (1:10 dilution). D. Aliquots of the supernatant obtained at different time points during the cultivation of P. pastoris expressing the wtgalox gene from F. graminearum without the N-terminal His10-tag extracellularly at 30°C; lane 1, molecular mass standard; lane 2-7, 12 h; 24 h; 48 h; 72 h; 96 h; 120 h.

Back to article page