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Strategies to display heterologous proteins on the cell surface of lactic acid bacteria using as anchor the C-terminal domain of Lactobacillus acidophilus SlpA

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Abstract

The surface-layer (S-layer) protein of Lactobacillus acidophilus is a crystalline array of self-assembling subunits, non-covalently bound to the most outer cell wall envelope, which constitutes up to 20% of the total cell protein content. These attributes make S-layer proteins an excellent anchor for the development of microbial cell-surface display systems. In L. acidophilus, the S-layer is formed predominantly by the protein SlpA. We have previously shown that the C-terminal domain of SlpA is responsible for the cell wall anchorage on L. acidophilus ATCC 4356. In the present study, we evaluated the C-terminal domain of SlpA of L. acidophilus ATCC 4356 as a potential anchor domain to display functional proteins on the surface of non-genetically modified lactic acid bacteria (LAB). To this end, green fluorescent protein (GFP)-CTSlpA was firstly produced in Escherichia coli and the recombinant proteins were able to spontaneously bind to the cell wall of LAB in a binding assay. GFP was successfully displayed on the S-layer stripped surface of L. acidophilus. Both the binding stability and cell survival of L. acidophilus decorated with the recombinant protein were then studied in simulated gastrointestinal conditions. Furthermore, NaCl was tested as a safer alternative to LiCl for S-layer removal. This study presents the development of a protein delivery platform involving L. acidophilus, a microorganism generally regarded as safe, which utilizes the contiguous, non-covalently attached S-layer at the cell surface of the bacterium without introducing any genetic modification.

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Funding

This work was supported by Grants from the Universidad de Buenos Aires (UBA) (20020170200329BA) and FONCyT (PICT 1337–2017), Argentina, to MMP, as well as by a doctoral Fellowship of the National Scientific and Technical Research Council (CONICET) to TBG, and an UBA undergraduate Fellowship to MCP. MCA, DFP, SMR, and MMP are members of CONICET.

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Authors and Affiliations

  1. Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Cdad. Universitaria, Pabellón II, 4 piso, Lab QB40, CABA, C1428EGA, Buenos Aires, Argentina

    Tania B. Gordillo, Miranda C. Palumbo, Mariana Claudia Allievi, Darío A. Fernández Do Porto, Sandra M. Ruzal & María Mercedes Palomino

  2. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales (IQUIBICEN), CONICET-Universidad de Buenos Aires, Buenos Aires, Argentina

    Tania B. Gordillo, Mariana Claudia Allievi, Sandra M. Ruzal & María Mercedes Palomino

  3. Facultad de Ciencias Exactas y Naturales, Instituto de Cálculo, Universidad de Buenos Aires, Buenos Aires, Argentina

    Darío A. Fernández Do Porto

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  1. Tania B. Gordillo
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  2. Miranda C. Palumbo
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  3. Mariana Claudia Allievi
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  4. Darío A. Fernández Do Porto
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  5. Sandra M. Ruzal
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  6. María Mercedes Palomino
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Correspondence to María Mercedes Palomino.

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Gordillo, T.B., Palumbo, M.C., Allievi, M.C. et al. Strategies to display heterologous proteins on the cell surface of lactic acid bacteria using as anchor the C-terminal domain of Lactobacillus acidophilus SlpA. World J Microbiol Biotechnol 36, 169 (2020). https://doi.org/10.1007/s11274-020-02945-9

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