EPathBrick: A synthetic biology platform for engineering metabolic pathways in E. coli

Peng Xu, Amerin Vansiri, Namita Bhan, Mattheos A.G. Koffas*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

173 Scopus citations

Abstract

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 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 can 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 (̃9 Kb) 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.

Original languageEnglish
Pages (from-to)256-266
Number of pages11
JournalACS Synthetic Biology
Volume1
Issue number7
DOIs
StatePublished - 20 Jul 2012
Externally publishedYes

Keywords

  • Gene assembly
  • Metabolic engineering
  • Pathway configuration
  • T7 promoter activity
  • Transcriptional fine-tuning

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