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Fig. 4 | BMC Biotechnology

Fig. 4

From: A hyper-thermostable α-amylase from Pyrococcus furiosus accumulates in Nicotiana tabacum as functional aggregates

Fig. 4

Purification of PFA from plant extract by electrophoresis with a preparative PAGE cell (a and b) and by heating (c). a GelCode blue-stained PAGE gel (top panel) and zymogram (bottom panel). Ten ml fractions were collected every 8 min for 8 h. Numbers denote the fraction number. Each lane was loaded with 20 μl sample. b Gelcode blue-stained PAGE loaded with crude extract and purified PFA from pooled fractions 45–55 which contained most PFA and very few other proteins. The pooled sample was concentrated and buffer-exchanged into reducing extraction buffer using a spin column with 10 kDa cut-off pore size. c Reduced extract was heated at different temperatures for 5 min and cleared by centrifugation. The supernatant was separated by SDS-PAGE. Top panel: zymogram. Lower panel: GelCode blue stained gel

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