A Coculture Based Tyrosine-Tyrosinase Electrochemical Gene Circuit for Connecting Cellular Communication with Electronic Networks

ACS Synth Biol. 2020 May 15;9(5):1117-1128. doi: 10.1021/acssynbio.9b00469. Epub 2020 Apr 2.

Abstract

There is a growing interest in mediating information transfer between biology and electronics. By the addition of redox mediators to various samples and cells, one can both electronically obtain a redox "portrait" of a biological system and, conversely, program gene expression. Here, we have created a cell-based synthetic biology-electrochemical axis in which engineered cells process molecular cues, producing an output that can be directly recorded via electronics-but without the need for added redox mediators. The process is robust; two key components must act together to provide a valid signal. The system builds on the tyrosinase-mediated conversion of tyrosine to L-DOPA and L-DOPAquinone, which are both redox active. "Catalytic" transducer cells provide for signal-mediated surface expression of tyrosinase. Additionally, "reagent" transducer cells synthesize and export tyrosine, a substrate for tyrosinase. In cocultures, this system enables real-time electrochemical transduction of cell activating molecular cues. To demonstrate, we eavesdrop on quorum sensing signaling molecules that are secreted by Pseudomonas aeruginosa, N-(3-oxododecanoyl)-l-homoserine lactone and pyocyanin.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • 4-Butyrolactone / analogs & derivatives
  • 4-Butyrolactone / analysis
  • 4-Butyrolactone / pharmacology
  • Culture Media / chemistry
  • Electrochemical Techniques
  • Electrodes
  • Gold / chemistry
  • Levodopa / chemistry
  • Levodopa / metabolism
  • Monophenol Monooxygenase / genetics
  • Monophenol Monooxygenase / metabolism*
  • Oxidation-Reduction
  • Plasmids / genetics
  • Plasmids / metabolism
  • Pseudomonas aeruginosa / genetics
  • Pseudomonas aeruginosa / metabolism
  • Pyocyanine / analysis
  • Pyocyanine / pharmacology
  • Quorum Sensing / drug effects
  • Synthetic Biology / methods*
  • Tyrosine / chemistry
  • Tyrosine / metabolism*

Substances

  • Culture Media
  • Tyrosine
  • Levodopa
  • Gold
  • N-(3-oxohexanoyl)-3-aminodihydro-2(3H)-furanone
  • Pyocyanine
  • Monophenol Monooxygenase
  • 4-Butyrolactone