Figure 4.

High producer selection pipeline. Combinatorial libraries were cloned and introduced into the production strains. Cultures carrying the pooled libraries were grown and expression was induced. Cells showing the highest GFP levels were selected by FACS, resulting in a cell suspension in PBS. The cell suspension was concentrated by filtration, transferred into a flask with fresh medium and grown overnight. The resulting culture was plated on agar before transferring 96 single colonies to a 96-deep-well plate. Expression was induced for 4 h. Cells were pelleted, the soluble protein fractions were extracted, and the GFP signal was quantified. The clones showing the most intense fluorescence were cultivated in flasks and subsequently analyzed by SDS-PAGE, western blot, and Sanger sequencing. Image created with BioRender.