The highest biocatalytic performance is achieved with glycerol as electron donor in combination with 5 mM phenylacetone and 37°C for biotransformation. (A) Effect of externally added carbohydrates as source of reducing power. Phenylacetone was converted by Top10 cells expressing PAMO in the presence of glucose, glycerol, or succinate after which the benzyl acetate content was analyzed. (B) Analysis of the optimal substrate concentration. Top10 cells producing PAMO were used for the conversion of the indicated amounts of phenylacetone followed by analysis of benzyl acetate production. (C) Influence of biotransformation temperature. Phenylacetone was converted by Top10 cells expressing PAMO at 25°C, 30°C, or 37°C followed by analysis of the benzyl acetate content. (D) Assessment of best biotransformation period. Top10 cells producing PAMO were used for the bioconversion of phenylacetone and the formation of benzyl acetate was analyzed at 1 hour intervals. For all panels, cells were grown as described in the legend to Figure 2A, and biotransformations were performed as described in the legend to Figure 1. All experiments were performed in duplicate and the values obtained were within 5% of each other.