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Title: Modified biosensors and biocatalysts and methods of use

Patent ·
OSTI ID:2222221

Biosensors including a nucleic acid encoding a PcaU protein, a PobR protein, a CatM protein, a PcaR protein, or a TphR protein are provided. In some examples, the biosensors include a promoter regulated by the sensed molecule operably linked to a reporter gene. The biosensors may be included in a vector or in cells including one or more of the biosensors or vectors. Modified chorismate pyruvate lyase (UbiC) and modified paraoxonase (PON1) proteins including one or more amino acid substitutions are provided. Finally, methods of selecting biocatalysts with increased activity including transforming a library of cells expressing a biosensor with one or more nucleic acids encoding one or more mutations in a gene involved in a biosynthesis pathway, determining activity of the reporter protein; and selecting a cell with increased reporter protein activity as expressing a biocatalyst with increased activity are provided.

Research Organization:
Los Alamos National Laboratory (LANL), Los Alamos, NM (United States)
Sponsoring Organization:
USDOE National Nuclear Security Administration (NNSA); USDOE Office of Energy Efficiency and Renewable Energy (EERE), Office of Sustainable Transportation. Bioenergy Technologies Office (BETO)
DOE Contract Number:
89233218CNA000001
Assignee:
Triad National Security, LLC (Los Alamos, NM)
Patent Number(s):
11,739,318
Application Number:
16/226,474
OSTI ID:
2222221
Resource Relation:
Patent File Date: 12/19/2018
Country of Publication:
United States
Language:
English

References (10)

Rosetta comparative modeling for library design: Engineering alternative inducer specificity in a transcription factor: A Comparative Approach for Protein Design journal May 2015
Biosensor-based engineering of biosynthetic pathways journal December 2016
Nucleic acid and amino acid sequences relating to Acinetobacter baumannii for diagnostics and therapeutics patent May 2003
Engineering an Acinetobacter regulon for biosensing and high-throughput enzyme screening in E. coli via flow cytometry journal May 2014
A microbial sensor for organophosphate hydrolysis exploiting an engineered specificity switch in a transcription factor journal August 2016
Evolution-guided optimization of biosynthetic pathways journal December 2014
Isolated PON1 polypeptides, polynucleotides encoding same and uses thereof in treating or preventing organophosphate exposure associated damage patent May 2014
Enhanced stereoselective hydrolysis of toxic organophosphates by directly evolved variants of mammalian serum paraoxonase: Enhanced stereoselective OP hydrolysis by PON1 journal April 2006
Directed evolution of mammalian paraoxonases PON1 and PON3 for bacterial expression and catalytic specialization journal December 2003
Characterization and Modeling of Transcriptional Cross-Regulation in Acinetobacter baylyi ADP1 journal June 2012

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