9806774 Khosla The overall goal of this project is to use the multi-enzyme polyketide synthases (PKSs) for catalyzing the enzymatic synthesis of polyketides using simple commercially available building blocks. Three major obstacles need to be overcome in order to accomplish this goal. 1) An inexpensive source of the component enzymes needs to be developed. Solutions to this problem will primarily rely on the power of recombinant DNA methods. 2) The stability of the PKSs must be maximized without significantly impacting quality with respect to turnover parameters and selectivity. This is to be accomplished through a combination of protein purification, optimization of reaction conditions (e.g., temperature, pH, and buffer composition), and protein immobilization. 3) An inexpensive source of substrates needs to be developed. Since PKSs utilize expensive cofactors such as NAD (P) H and CoA derivatives, this problem is to be addressed through cofactor regeneration and/or the development of simpler substrate mimics that can be accepted by PKSs. ***
