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Optimization of recombinant BsDb expression in Pichia pastoris. Supernatants collected at each evaluated condition were analyzed using ELISA and spot Western blotting analysis. (A) Optimization of the methanol inducing time; (B) optimization of the methanol concentration; (C) optimization of the pH value; and (D) optimization of the cell density. The experiment was performed in triplicate, and the mean values ± SD were presented.
