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Dynamic monitoring of oscillatory enzyme activity of individual live bacteria via nanoplasmonic optical antennas

Nature Photonics volume 17, pages 904–911 (2023)Cite this article

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Abstract

Outer membrane vesicles (OMVs) are extracellular structures derived from the outer membrane of bacteria. They carry diverse cargos such as proteins, nucleic acids and enzymes, which are released by bacteria to communicate with each other and with host cells. Understanding the role of OMVs as carriers of signalling enzymes provides insights into intercellular communication, pathogenesis and biofilm formation, among others. Although fluorescence-based techniques have been employed to study vesicles, real-time live monitoring of enzyme-based cellular communication has remained challenging due to undesired effects of photobleaching and interference from labelling agents. Here we report label-free dynamic monitoring of the oscillatory activity of the enzyme azoreductase (AzoR) in individual live bacteria via nanoplasmonic optical antennas. Our nanoplasmonic antennas consist of gold nanorods modified with black hole quencher molecules (BHQ-3), whose scattering cross-section is modulated by the presence of AzoR. The antennas enable long-term (several hours) and distance-dependent (up to 3 μm) detection of AzoR via OMVs released by individual live bacteria. We observe periodic oscillatory enzyme activity in different living environments and at different stages of bacterial growth. We also found that oscillatory enzyme activity exhibits heterogeneous features due to the coupling of oscillation signals between neighbouring bacteria. The dynamic monitoring of signalling enzymes paves the way for a better understanding of mechanisms of bacterial communication, pathogenesis and drug resistance.

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Fig. 1: Understanding bacterial cell–cell communication via the observation of oscillatory enzyme activity by innovative biological sensing mechanism of nanoplasmonic optical antennas.
Fig. 2: Distance-dependent detection of enzyme release from a single bacterium by nanoplasmonic optical antennas.
Fig. 3: Real-time detection of oscillatory enzyme activity of a single bacterium.
Fig. 4: Real-time detection of oscillation coupling of enzyme activity in communicating single bacteria.

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Data availability

All of the data that support the findings of this study are reported in the main text and Supplementary Information. Source Data are available from the corresponding authors upon reasonable request.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (grant nos. 62135005, 61975065, 32271405 and 12204196 to B.L., H.X., T.P. and Y.S., respectively), Guangdong Basic and Applied Basic Research Foundation (grant no. 2022B1515120012 to H.X.) and Science and Technology Program of Guangzhou (grant no. 202201010370 to T.P.).

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Authors and Affiliations

  1. Guangdong Provincial Key Laboratory of Nanophotonic Manipulation, Institute of Nanophotonics, Jinan University, Guangzhou, China

    Dengyun Lu, Guoshuai Zhu, Xing Li, Jianyun Xiong, Danning Wang, Yang Shi, Ting Pan, Baojun Li & Hongbao Xin

  2. Renal Division and Division of Engineering in Medicine, Department of Medicine, Brigham and Women’s Hospital, Harvard Medical School, Harvard University, Boston, MA, USA

    Luke P. Lee

  3. Department of Bioengineering, Department of Electrical Engineering and Computer Science, University of California at Berkeley, Berkeley, CA, USA

    Luke P. Lee

  4. Institute of Quantum Biophysics, Department of Biophysics, Sungkyunkwan University, Suwon, Korea

    Luke P. Lee

  5. Department of Chemistry and Nanoscience, Ewha Womans University, Seoul, Korea

    Luke P. Lee

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  1. Dengyun Lu
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Contributions

H.X. and L.P.L. conceptualized the work, D.L., B.L. and H.X. designed the study. D.L., G.Z., D.W., J.X. and T.P. performed the experiments. Y.S. and X.L. performed simulations. D.L., B.L., H.X. and L.P.L. wrote the paper. All the authors contributed to the data analysis, discussion and manuscript preparation.

Corresponding authors

Correspondence to Baojun Li, Luke P. Lee or Hongbao Xin.

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Nature Photonics thanks Shaopeng Wang and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary Figs. 1–25, Notes 1–4 and refs. 1–9.

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Video showing the measurement for a single bacterium.

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Lu, D., Zhu, G., Li, X. et al. Dynamic monitoring of oscillatory enzyme activity of individual live bacteria via nanoplasmonic optical antennas. Nat. Photon. 17, 904–911 (2023). https://doi.org/10.1038/s41566-023-01265-2

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