Figure 3
From: Alanine substitution in cellobiohydrolase provides new insights into substrate threading
Production of wild-type (WT) and nine alanine-substituted mutants of TcCel7A. (a) SDS-PAGE of WT and nine mutants. NC: untransformed A. oryzae strain NS4DLP. BSA concentrations: 10, 50, 100, and 200 mg/L. Target CBHs are shown in a dashed square and the α-amylase position is denoted with a wedge. Full-length gels are presented in Supplementary Fig. S2. (b) Western blotting of WT, K203A, W63A, W65A, and N62A. Dilution ratios follow the name for each sample. All images are excerpts from an identical membrane (Supplementary Fig. S3). (c) Pseudo-coloured SDS-PAGE images of N62A from four different cultures and of WT with serial dilutions (×2–64) for comparison. For each lane, concentration was determined using image densitometry. Full-length gels with original colour are presented in Supplementary Fig. S4.
