Fig. 7

The expression and purification of recombianant Man1E analyzed by SDS-PAGE. a Recombinant Man1E analyzed by SDS-PAGE. Lane 1: Precipitation after cell lysis from Escherichia coli BL21(DE3) harboring pET28a (+); Lane 2: Cell lysate from Escherichia coli BL21(DE3) harboring pET28a (+); Lane 3: Fermentation supernatant from Escherichia coli BL21(DE3) harboring pET28a (+); Lane M:Marker; Lane 4: Precipitation after cell lysis from Escherichia coli BL21(DE3) harboring pET28a (+)-Man1E; Lane 5: Cell lysate from Escherichia coli BL21(DE3) harboring pET28a (+)-Man1E; Lane 6: Fermentation supernatant from Escherichia coli BL21(DE3) harboring pET28a (+)-Man1E. b SDS–PAGE analysis of purified Man1E. Lane 1: Supernatant after cell breakage from Escherichia coli BL21(DE3) harboring pET28a (+); Lane 2: Supernatant after cell breakage from Escherichia coli BL21(DE3) harboring pET28a (+)-Man1E; Lane 3: Purified recombinanant Man1E; Lane M: Marker. c-e Optimization of soluble expression for recombianant Man1E. Induction temperature (c). Lane M: Marker; Lane 1: 16 °C; Lane 2: 20 °C; Lane 3: 24 °C; Lane 4: 28 °C; Lane 5: 32 °C; Lane 6: 37 °C; Lane 7: 42 °C. PTG concentration (d). Lane M: Marker; Lane 1: 0.2 mM; Lane 2: 0.4 mM; Lane 3: 0.6 mM; Lane 4: 0.8 mM; Lane 5: 1.0 mM; Lane 6: 1.2 mM. Induction time (c). Lane 1: 4 h; Lane 2: 8 h; Lane 3: 12 h; Lane 4: 16 h. The band shown by the arrow was recombinant Man1E