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Title: A protocatechuate biosensor for Pseudomonas putida KT2440 via promoter and protein evolution

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. In this study, 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.
Authors:
 [1] ;  [1] ; ORCiD logo [2] ;  [1] ;  [2] ;  [1] ;  [1] ; ORCiD logo [2] ;  [1]
  1. Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
  2. National Renewable Energy Lab. (NREL), Golden, CO (United States). National Bioenergy Center
Publication Date:
Report Number(s):
NREL/JA-5100-71276; LA-UR-17-23446
Journal ID: ISSN 2214-0301
Grant/Contract Number:
AC36-08GO28308; NL0032182; CBCALL12-LS-6-0622; AC52-06NA25396
Type:
Accepted Manuscript
Journal Name:
Metabolic Engineering Communications
Additional Journal Information:
Journal Volume: 6; Journal Issue: C; Journal ID: ISSN 2214-0301
Publisher:
Elsevier
Research Org:
National Renewable Energy Lab. (NREL), Golden, CO (United States); Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
Sponsoring Org:
USDOE Office of Energy Efficiency and Renewable Energy (EERE), Bioenergy Technologies Office (EE-3B); Defense Threat Reduction Agency (DTRA)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; whole cell biosensor; aromatic catabolism; transcription factor; PcaU; Shikimate; Biological Science
OSTI Identifier:
1432606
Alternate Identifier(s):
OSTI ID: 1438356