Figure 1

Top10 is the best expression host in combination with 0.2% L-arabinose and 30°C for the production of PAMO. The expression of PAMO was investigated in the standard expression strains Top10, MC1061, and BL21(DE)3 transformed with an arabinose-inducible PAMO expression plasmid. Cells were grown in the presence of increasing amounts of inducer (L-arabinose) in 96-sdMTP over night at 25°C (A), 30°C (B), or 37°C (C), respectively. The next day, cells were harvested and used for the bioconversion of phenylacetone in 96-sdMTP for 3 hours at 37°C followed by the analysis of the benzyl acetate content. All experiments were performed in duplicate and the values obtained were within 5% of each other. (D) PAMO expression in all tested strains. Cells were grown as described in the legend to Figure 1A at 30°C, or 37°C, respectively. Subsequently, a cell lysate was prepared, which was further separated into a soluble (sol) and insoluble fraction (insol) for Top10 and MC1061 by ultracentrifugation. Samples were analyzed by SDS-PAGE and Coomassie staining. The position of PAMO is indicated by an asterisk.