Article luxab-based microbial cell factories for the sensing, manufacturing and transformation of industrial aldehydes

Thomas Bayer, Aileen Becker, Henrik Terholsen, In Jung Kim, Ina Menyes, Saskia Buchwald, Kathleen Balke, Suvi Santala, Steven C. Almo, Uwe T. Bornscheuer

Research output: Contribution to journalArticlepeer-review

Abstract

The application of genetically encoded biosensors enables the detection of small molecules in living cells and has facilitated the characterization of enzymes, their directed evolution and the engineering of (natural) metabolic pathways. In this work, the LuxAB biosensor system from Photorhabdus luminescens was implemented in Escherichia coli to monitor the enzymatic production of aldehydes from primary alcohols and carboxylic acid substrates. A simple high-throughput assay utilized the bacterial luciferase—previously reported to only accept aliphatic long-chain aldehydes—to detect structurally diverse aldehydes, including aromatic and monoterpene aldehydes. LuxAB was used to screen the substrate scopes of three prokaryotic oxidoreductases: an alcohol dehydrogenase (Pseudomonas putida), a choline oxidase variant (Arthrobacter chlorophenolicus) and a carboxylic acid reductase (Mycobacterium marinum). Consequently, high-value aldehydes such as cinnamaldehyde, citral and citronellal could be produced in vivo in up to 80% yield. Furthermore, the dual role of LuxAB as sensor and monooxygenase, emitting bioluminescence through the oxidation of aldehydes to the corresponding carboxylates, promises implementation in artificial enzyme cascades for the synthesis of carboxylic acids. These findings advance the bio-based detection, preparation and transformation of industrially important aldehydes in living cells.

Original languageEnglish (US)
Article number953
JournalCatalysts
Volume11
Issue number8
DOIs
StatePublished - Aug 2021

Keywords

  • Aldehyde production
  • Bioluminescence
  • Biosensor
  • Enzyme cascade
  • High-throughput screening
  • Luciferase
  • Whole-cell biocataly-sis

ASJC Scopus subject areas

  • Catalysis
  • Physical and Theoretical Chemistry

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