Design and standardization of synthetic biofluorination pathways
WP2 is composed by 3 tasks that will result in the establishment of a standardized, efficient synthetic biofluorination pathway from SAM to fluoroacetaldehyde. Genes encoding the 4-step fluorination pathway of Streptomyces cattleya and other Streptomyces species will be optimized in silico and tested in wild-type P. putida. WP2 also validates a set of standardized FluoroBricks (encoding the synthetic biofluorination pathway) both from cell-free extracts of P. putida after multiple gene expression and by reconstructing the biosynthesis of individual fluorometabolites via combining optimized enzymes in vitro.