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Carbon nanotube uptake in cyanobacteria for near-infrared imaging and enhanced bioelectricity generation in living photovoltaics

Nature Nanotechnology volume 17, pages 1111–1119 (2022)Cite this article

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Abstract

The distinctive properties of single-walled carbon nanotubes (SWCNTs) have inspired the development of many novel applications in the field of cell nanobiotechnology. However, studies thus far have not explored the effect of SWCNT functionalization on transport across the cell walls of prokaryotes. We explore the uptake of SWCNTs in Gram-negative cyanobacteria and demonstrate a passive length-dependent and selective internalization of SWCNTs decorated with positively charged biomolecules. We show that lysozyme-coated SWCNTs spontaneously penetrate the cell walls of a unicellular strain and a multicellular strain. A custom-built spinning-disc confocal microscope was used to image the distinct near-infrared SWCNT fluorescence within the autofluorescent cells, revealing a highly inhomogeneous distribution of SWCNTs. Real-time near-infrared monitoring of cell growth and division reveal that the SWCNTs are inherited by daughter cells. Moreover, these nanobionic living cells retained photosynthetic activity and showed an improved photo-exoelectrogenicity when incorporated into bioelectrochemical devices.

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Fig. 1: NIR imaging of internalized SWCNTs.
Fig. 2: Localization of internalized SWCNTs.
Fig. 3: Effect of surface charge on SWCNT uptake.
Fig. 4: SWCNT internalization by strains lacking T4P.
Fig. 5: Effect of SWCNT internalization on photosynthesis and cell viability.
Fig. 6: Biophotovoltaic and electrode characterization of nanobionic Synechocystis cells.

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Data availability

Data supporting the findings of this study are available within the paper and Supplementary Information. Raw data (microscopy, spectroscopy and electrochemical measurements) of the results reported in this study are available via Zenodo at https://doi.org/10.5281/zenodo.6790098. All other relevant data and findings of this study are available from the corresponding author upon reasonable request.

Code availability

Details of the custom MATLAB code are available in the Supplementary Information. The complete custom MATLAB code is available via Zenodo at https://doi.org/10.5281/zenodo.6777770.

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Acknowledgements

We thank the Swiss National Science Foundation (SNSF) for support under an Assistant Professor (AP) Energy Grant (Project No. PYAPP2_154269) (A.A.B) and Swiss National Science Foundation Project No. IZLIZ2_182972 (A.A.B.). We also thank K. Sivula from the Laboratory for Molecular Engineering of Optoelectronic Nanomaterials (LIMNO) at EPFL for access to the AFM facility.

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Authors and Affiliations

  1. Institute of Chemical Sciences and Engineering (ISIC), Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland

    Alessandra Antonucci, Melania Reggente, Charlotte Roullier, Alice J. Gillen, Nils Schuergers, Vitalijs Zubkovs, Benjamin P. Lambert, Mohammed Mouhib & Ardemis A. Boghossian

  2. Institute of Biology III, University of Freiburg, Freiburg, Germany

    Nils Schuergers

  3. Swiss Center for Electronics and Microtechnology (CSEM), Landquart, Switzerland

    Vitalijs Zubkovs

  4. Department of Biological and Environmental Sciences and Technologies (DiSTeBA), University of Salento, Lecce, Italy

    Elisabetta Carata

  5. Department of Biology and Biotechnology Charles Darwin, Sapienza University of Rome, CNR Nanotec, Lecce, Italy

    Luciana Dini

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  1. Alessandra Antonucci
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Contributions

A.A. and A.A.B conceived the project and designed the study. A.A., M.R., and C.R. performed cell imaging and uptake experiments. V.Z. built the NIR confocal microscope used in the study. M.R. and C.R. performed the electrochemical measurements. M.R. performed the AFM imaging. B.P.L. prepared the samples used for the density-gradient centrifugation and helped with the fluorescence imaging experiments. M.M. performed the western blot analysis and N.S. helped with the biological sample preparation. A.J.G. helped with the analysis of the fluorescence measurements. L.D. and E.C. designed the TEM experiments, and E.C. performed the sample preparation and TEM imaging. A.A.B. analysed the collected data. All authors contributed to the preparation, writing and editing of this manuscript.

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Correspondence to Ardemis A. Boghossian.

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Nature Nanotechnology thanks Sebastian Kruss, Khaled Selim and Christa Walther for their contribution to the peer review of this work.

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Supplementary Figs. 1–25, Tables 1–3, Methods, Discussion and references.

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Antonucci, A., Reggente, M., Roullier, C. et al. Carbon nanotube uptake in cyanobacteria for near-infrared imaging and enhanced bioelectricity generation in living photovoltaics. Nat. Nanotechnol. 17, 1111–1119 (2022). https://doi.org/10.1038/s41565-022-01198-x

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