Advanced biofuels produced by microbes have similar properties to petroleum-based fuels, and can ‘drop in’ to the existing transportation infrastructure.  Two main challenges to the production of advanced biofuels in high yields are the engineering of the microbe’s metabolism and the optimization of rate limiting enzymes in key metabolic pathways.  In this seminar, I will discuss synthetic biology and protein engineering approaches to address these two challenges.  First, I will present the metabolic engineering of microbes for the production of advanced biofuels that can serve as replacements to diesel and jet fuel.  Second, using structural information of key enzymes in the pathway, I will discuss how we try to engineer the enzymes to overcome their limitations and reach higher microbial titers of the advanced biofuel.  These projects illustrate an interdisciplinary approach to cell engineering that combines chemistry, genetics, enzyme and metabolic engineering, and synthetic biology for the production of biofuels.