Discovery and evolution of biologically active metabolites

A method for the discovery and evolution of metabolic pathways that produce molecules that modulate protein activity, comprising:

• contacting a plurality of host cells that comprise a protein of interest with a plurality of expression vectors that each comprise a different metabolic pathway of a plurality of metabolic pathways under conditions sufficient to express components of the plurality of metabolic pathways in the plurality of host cells, wherein each metabolic pathway of the plurality of metabolic pathways produces one or more metabolites, wherein the protein of interest comprises an enzyme, wherein the enzyme comprises a protein tyrosine phosphatase;
• expressing the plurality of metabolic pathways in the plurality of host cells, wherein a host cell of the plurality of host cells or a subset of the plurality of host cells produces a detectable output when a metabolic pathway of the plurality of metabolic pathways produces one or more products comprising a metabolite of the one or more metabolites that modulates an activity of the enzyme, wherein the activity of the enzyme controls an assembly of a protein complex, and wherein assembly of the protein complex enhances transcription of a gene of interest to produce the detectable output;
• screening the host cell or the subset of the plurality of host cells under conditions that enable measurement of the detectable output in the host cell or the subset of the plurality of host cells;
• isolating the host cell or the subset of the plurality of host cells that produce the detectable output;
• analyzing an expression vector of the plurality of expression vectors that yields a detectable output that is higher than an output of a reference vector comprising a reference pathway that causes a threshold level of transcription of the gene of interest;
• and
• characterizing the one or more products of metabolic pathways of the plurality of metabolic pathways comprised by the expression vector that yielded the detectable output that is higher than the output of the reference vector.