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Figure 3 | Microbial Cell Factories

Figure 3

From: Identification of a heterologous cellulase and its N-terminus that can guide recombinant proteins out of Escherichia coli

Figure 3

Cell lysis determination and inactive Cel-CD secretion analysis. (A) Immunoblotting detection of Cel-CD, GroEL and MBP in the culture medium. Samples were subjected to immunoblotting with an anti-Cel-CD antibody, an anti-GroEL antibody or an anti-MBP antibody. (B) SDS analysis of cellulase (wt) and its mutants (E160Q, E160Q&E274Q) secreted into the culture medium. The activities of the secreted mutants of Cel-CD (E160Q, E160Q&E274Q) were analyzed by Congo red staining method (4 h, 6 h, 8 h, 16 h). (C) TEM images of E. coli BL21 (DE3) that containing different plasmids. (1) pET28a/cel, (2) pET28a/E160Q, (3) pET28a/E160Q&E274Q, (4) pET28a. Protein samples were separated on 12% SDS-PAGE. Aliquots corresponding to 20 μL of the culture medium were loaded onto the gel. Molecular size markers are shown in kDa. Lane M is the marker; Lane C is the control of collected cells.

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