Fig. 2
Native PAGE/western blot analysis of supernatants from recombinant C. acetobutylicum strains producing heterologous, FLAG-tagged cellulosome components encoded by chromosomally integrated gene constructs. Supernatants were obtained at an OD600 of approximately 0.8 and proteins concentrated approximately 100-fold by ultrafiltration with a 10,000 MWCO filter before being separated on 3–8 % Tris–Glycine gradient gels; after blotting, cellulosomal components were detected using ANTI-FLAG M2 monoclonal antibody-horseradish peroxidise conjugate. Supernatants were obtained 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