Expanding the Synthetic Biology Toolbox for Engineering Metabolic Pathways in E. Coli

Expanding the Synthetic Biology Toolbox for Engineering Metabolic Pathways in E. coli

Harnessing cell factories for producing biofuel and pharmaceutical molecules has stimulated efforts to develop novel synthetic biology tools customized for modular pathway engineering and optimization. Here we report the development of a set of vectors compatible with BioBrick^TM standards and its application in metabolic engineering. The engineered ePathBrick vectors comprise four compatible restriction enzyme sites allocated on strategic positions so that different regulatory control signals can be reused and manipulation of expression cassette be streamlined. Specifically, these vectors allow for fine-tuning gene expression by integrating multiple transcriptional activation or repression signals into the operator region. At the same time, ePathBrick vectors support the modular assembly of pathway components and combinatorial generation of pathway diversities with three distinct configurations. We also demonstrated the functionality of a seven gene pathway (~9Kb) assembled on one single ePathBrick vector. The ePathBrick vectors presented here provide a versatile platform for rapid design and optimization of metabolic pathways in E. coli.

Keywords: metabolic engineering; transcriptional fine-tuning; gene assembly; pathway configuration; T7 promoter activity

Reference

Xu P, Vansiri A, Bhan N, and Koffas M.A.G. (2012) ePathBrick: A synthetic biology platform for engineering metabolic pathways in E. coli. ACS Synthetic Biology, DOI: 10.1021/sb300016b, In Press.