In this study, the Leloir pathway of galactose metabolism was rebuilt in Escherichia coil to remove CCR and amplify galactose utilization rate. All genes encoding pathway enzymes were expressed under the control of a synthetic module that included promoters, 5'-untranslated regions, and terminators as a re-organized single operon in the chromosome. The engineered strain showed both an enhanced galactose utilization rate and the capacity to simultaneously assimilate galactose and glucose. This work demonstrates the feasibility of using synthetic biology tools to re-build biological systems for engineering purpose. (C) 2012 Elsevier Ltd. All rights reserved.