Microbial production of biochemicals from renewable sources is a cost-effective alternative to chemical synthesis production methods using petroleum-based feedstocks. The efficiency of microbial production processes is often enhanced by altering metabolism using metabolic engineering concepts and strategies. In this work, a strategy is developed to enhance the production of pyruvate-derived biochemicals by modifying pyruvate dehydrogenase, a key enzyme in pyruvate metabolism. The first study provides proof-of-concept for this strategy by generating pyruvate dehydrogenase variants that influence metabolic flux at the pyruvate node in central metabolism. The second study demonstrates that pyruvate dehydrogenase variants enhance the production of acetoin, a pyruvate-derived biochemical. The final study broadens the substrate range for production of pyruvate or pyruvate-derived biochemicals by engineering Escherichia coli strains able to efficiently produce pyruvate from sucrose.