A Protocatechuate Biosensor for Pseudomonas putida KT2440 via Promoter and Protein Evolution

Christopher Johnson, Gregg Beckham, Ramesh Jha, Jeremy Bingen, Theresa Kern, Payal Khanna, Daniel Trettel, Charlie Strauss, Taraka Dale

Research output: Contribution to journalArticlepeer-review

28 Scopus Citations

Abstract

Robust fluorescence-based biosensors are emerging as critical tools for high-throughput strain improvement in synthetic biology. Many biosensors are developed in model organisms where sophisticated synthetic biology tools are also well established. However, industrial biochemical production often employs microbes with phenotypes that are advantageous for a target process, and biosensors may fail to directly transition outside the host in which they are developed. In particular, losses in sensitivity and dynamic range of sensing often occur, limiting the application of a biosensor across hosts. Here we demonstrate the optimization of an Escherichia coli-based biosensor in a robust microbial strain for the catabolism of aromatic compounds, Pseudomonas putida KT2440, through a generalizable approach of modulating interactions at the protein-DNA interface in the promoter and the protein-protein dimer interface. The high-throughput biosensor optimization approach demonstrated here is readily applicable towards other allosteric regulators.

Original languageAmerican English
Pages (from-to)33-38
Number of pages6
JournalMetabolic Engineering Communications
Volume6
DOIs
StatePublished - 2018

Bibliographical note

Publisher Copyright:
© 2018

NREL Publication Number

  • NREL/JA-5100-71276

Keywords

  • Aromatic catabolism
  • PcaU
  • Shikimate
  • Transcription factor
  • Whole cell biosensor

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