Figure 5From: Secretion and assembly of functional mini-cellulosomes from synthetic chromosomal operons in Clostridium acetobutylicum ATCC 824Expression of C. thermocellum cellulases in C. acetobutylicum. (a) Recombinant C. acetobutylicum strains expressing the Cel8A, Cel9B and Cel9K cellulases were grown until late stationary phase (OD600 > 3) and their supernatants TCA precipitated, concentrated 100-fold and analysed by SDS-PAGE. (b) The recombinant strain expressing the Cel48S was grown in buffered 2xYTG until early exponential phase (OD600 of 0.4[1] and 0.8[2]), concentrated 100-fold and separated by SDS-PAGE. All four cellulases were detected using ANTI-FLAG M2 monoclonal antibody-horseradish peroxidise conjugate (Sigma). Expected molecular masses are indicated by arrows. M, ColorPlus Prestained Protein Ladder (10–230 kDa; New England Biolabs), Flag, N-Flag-Bap control protein (50 kDa, 50 ng/lane; Sigma).Back to article page