Identification and Engineering of Transporters for Efficient Melatonin Production in Escherichia coli

Lei Yang*, Sailesh Malla, Emre Özdemir, Se Hyeuk Kim, Rebecca Lennen, Hanne B. Christensen, Ulla Christensen, Lachlan J. Munro, Markus J. Herrgård, Douglas B. Kell*, Bernhard Palsson

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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Abstract

Transporter discovery and engineering play an important role in cell factory development. Decreasing the intracellular concentration of the product reduces product inhibition and/or toxicity. Lowering intracellular concentrations is especially beneficial for achieving a robust strain at high titers. However, the identification of transporters for xenobiotic chemicals in the host strain is challenging. Here we present a high-throughput workflow to discover Escherichia coli transporters responsible for the efflux of the inhibitory xenobiotic compound melatonin. We took advantage of the Keio collection and screened about 400 transporter knockouts in the presence of a high concentration of melatonin. We found five transporters that when knocked out showed decreased tolerance to melatonin, indicating they are exporters of melatonin. We overexpressed these five genes individually in the production strain and found that one of them, yhjV, encoding a transporter with unknown substrates, resulted in a 27% titer increase in cultivation mimicking fed-batch fermentation. This study demonstrates how microbial cell factories can be improved through transporter identification and engineering. Further, these results lay the foundation for the scale-up of melatonin production in E. coli.

Original languageEnglish
Article number880847
JournalFrontiers in Microbiology
Volume13
Number of pages10
ISSN1664-302X
DOIs
Publication statusPublished - 2022

Keywords

  • E. coli
  • Identification
  • Inhibition
  • Membrane
  • Production
  • Screening
  • Toxicity
  • Transporter

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