Utilizing a Cell-free Protein Synthesis Platform for Natural Product Synthesis

Natural products are a great source of pharmaceuticals, providing a majority of all small molecule drugs that exist today. However, creating natural products through organic synthesis or in heterologous hosts can be difficult and time-consuming. Therefore, to allow for easier screening and production of natural products, we demonstrated the use of a cell-free protein synthesis (CFPS) system to partially assemble natural products in vitro using coupled enzyme reactions. The tea caffeine synthase TCS1 was utilized to synthesize caffeine within a CFPS system. Cell-free systems also provide the benefit of allowing the use of substrates that would normally be toxic in a cellular environment to synthesize novel products. The automation and reduced metabolic engineering requirements of CFPS systems combined with other synthesis methods can allow for the efficient generation of new compounds.