Figure 4

Expression profile and purification of chimeric CBHI protein in bioreactor fermentation of Y. lipolytica transformant. (A) Time course for the concentrated culture supernatant of Y. lipolytica[chimeric CBHI] transformant. The 25-fold concentrated supernatants (16 μL per well) were separated by SDS-PAGE, followed by Western blot analysis for the detection of chimeric CBHI. (B) SDS-PAGE and Western blot analyses for collected fractions during chimeric protein purification procedure. The chromatography fractions corresponding to gel lane numbers are: lane 1, the 20-fold concentrated culture supernatant (10 μL); lane 2, effluent from hydrophobic interaction chromatography (20 μL); lane 3, effluent from anion exchange column (20 μL); lanes 4 and 5, the minor impurity and peaks from size exclusion column (20 μL), respectively.