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Multiplexed assessment of engineered bacterial constructs for intracellular β-galactosidase expression by redox amplification on catechol-chitosan modified nanoporous gold

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Abstract

Synthetic biology approaches for rewiring of bacterial constructs to express particular intracellular factors upon induction with the target analyte are emerging as sensing paradigms for applications in environmental and in vivo monitoring. To aid in the design and optimization of bacterial constructs for sensing analytes, there is a need for lysis-free intracellular detection modalities that monitor the signal level and kinetics of expressed factors within different modified bacteria in a multiplexed manner, without requiring cumbersome surface immobilization. Herein, an electrochemical detection system on nanoporous gold that is electrofabricated with a biomaterial redox capacitor is presented for quantifying β-galactosidase expressed inside modified Escherichia coli constructs upon induction with dopamine. This nanostructure-mediated redox amplification approach on a microfluidic platform allows for multiplexed assessment of the expressed intracellular factors from different bacterial constructs suspended in distinct microchannels, with no need for cell lysis or immobilization. Since redox mediators present over the entire depth of the microchannel can interact with the electrode and with the E. coli construct in each channel, the platform exhibits high sensitivity and enables multiplexing. We envision its application in assessing synthetic biology-based approaches for comparing specificity, sensitivity, and signal response time upon induction with target analytes of interest.

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Acknowledgements

Support from University of Virginia’s Global Infectious Diseases Institute, from MOST (Taiwan) grant #107-2923-M-001-011-MY3 and from AFOSR grants FA2386-18-1-4100 and FA2386-21-1-4070 are acknowledged.

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Authors and Affiliations

  1. Electrical and Computer Engineering, University of Virginia, Charlottesville, VA, 22904, USA

    Yi Liu, John H. Moore & Nathan S. Swami

  2. 711th Human Performance Wing, Air Force Research Laboratory, Wright-Patterson Air Force Base, OH, 45433, Dayton, USA

    Svetlana Harbaugh & Jorge Chavez

  3. Institute of Physics, Academia Sinica, Taipei, Taiwan

    Chia-Fu Chou

  4. Chemistry, University of Virginia, Charlottesville, VA, 22904, USA

    Nathan S. Swami

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  1. Yi Liu
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Correspondence to Nathan S. Swami.

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Liu, Y., Moore, J.H., Harbaugh, S. et al. Multiplexed assessment of engineered bacterial constructs for intracellular β-galactosidase expression by redox amplification on catechol-chitosan modified nanoporous gold. Microchim Acta 189, 4 (2022). https://doi.org/10.1007/s00604-021-05109-0

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