(96g) 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.