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Fig. 1 | Biotechnology for Biofuels

Fig. 1

From: Production of a functional cell wall-anchored minicellulosome by recombinant Clostridium acetobutylicum ATCC 824

Fig. 1

SDS-PAGE/western blot analysis of supernatants from recombinant C. acetobutylicum strains producing heterologous cellulosome components encoded by chromosomally integrated gene constructs. Supernatants were obtained at an OD600 of approximately 0.8 and proteins concentrated 100-fold by TCA precipitation before being separated on 4–12 % Bis–Tris gradient gels. All cellulosomal components have been engineered to bear the FLAG epitope tag at the C-terminus, allowing visualisation using ANTI-FLAG M2 monoclonal antibody-horseradish peroxidise conjugate. Supernatants were obtained from wild-type C. acetobutylicum (WT) and from strains CEL11, expressing a gene encoding Cel9G-FLAG, CEL12, expressing a gene encoding Cel48F-FLAG, CEL13, expressing a gene encoding Xyn10A-FLAG, CEL15, expressing genes encoding all three enzymes (Xyn10A-FLAG, Cel9G-FLAG, and Cel48F-FLAG), CEL01, expressing a gene encoding the BBCipC3-FLAG miniscaffoldin, and CEL17, expressing genes encoding all three enzymes and the BBCipC3-FLAG miniscaffoldin. L, ColorPlus Prestained Protein Ladder (10–230 kDa); +, carboxy terminal FLAG-BAP control protein (50 kDa)

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