Abstract
This chapter describes the use of Lactococcus lactis as a safe and efficient cell factory to produce heterologous proteins of medical interest. The relevance of the use of this lactic acid bacterium (LAB) is that it is a noncolonizing, nonpathogenic microorganism that can be delivered in vivo at a mucosal level. The use of strains of L. lactis in clinical trials in humans to alleviate inflammatory bowel diseases has opened up the possibility of using this same LAB to target other diseases.
Several crucial aspects are addressed in this chapter, such as the expression of heterologous protein, subcellular compartment into which the heterologous protein is located, and description of a standardized protocol to process samples in cell and cell-free fractions to detect the targeted protein expressed by L. lactis.
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References
Wells, J. M., and Mercenier, A. (2008) Mucosal delivery of therapeutic and prophylactic molecules using lactic acid bacteria. Nat. Rev. Microbiol. 6, 349–362.
Van Huynegem, K., Loos, M., and Steidler, L. (2009) Immunomodulation by genetically engineered lactic acid bacteria. Front. Biosci. 14, 4825–4835.
de Ruyter, P. G., Kuipers, O. P., and de Vos, W. M. (1996) Controlled gene expression systems for Lactococcus lactis with the food-grade inducer nisin. Appl. Environ. Microbiol. 62, 3662–3667.
Steidler, L., Neirynck, S., Huyghebaert, N., Snoeck, V., Vermeire, A., Goddeeris, B., Cox, E., Remon, J. P., and Remaut, E. (2003) Biological containment of genetically modified Lactococcus lactis for intestinal delivery of human interleukin 10. Nat. Biotechnol. 21, 785–789.
Bermudez-Humaran, L. G., Langella, P., Cortes-Perez, N. G., Gruss, A., Tamez-Guerra, R. S., Oliveira, S. C., Saucedo-Cardenas, O., Montes de Oca-Luna, R., and Le Loir, Y. (2003) Intranasal immunization with recombinant Lactococcus lactis secreting murine interleukin-12 enhances antigen-specific Th1 cytokine production. Infect. Immun. 71, 1887–1896.
Steidler, L., Robinson, K., Chamberlain, L., Schofield, K. M., Remaut, E., Le Page, R. W., and Wells, J. M. (1998) Mucosal delivery of murine interleukin-2 (IL-2) and IL-6 by recombinant strains of Lactococcus lactis coexpressing antigen and cytokine. Infect. Immun. 66, 3183–3189.
Villatoro-Hernandez, J., Loera-Arias, M. J., Gamez-Escobedo, A., Franco-Molina, M., Gomez-Gutierrez, J. G., Rodriguez-Rocha, H., Gutierrez-Puente, Y., Saucedo-Cardenas, O., Valdes-Flores, J., and Montes-de-Oca-Luna, R. (2008) Secretion of biologically active interferon-gamma inducible protein-10 (IP-10) by Lactococcus lactis. Microb. Cell Fact. 7, 22.
Zavala-Flores, L. M., Villatoro-Hernandez, J., Gamez-Escobedo, A., Franco-Molina, M., Rangel-Colmenero, B. R., Villanueva-Olivo, A., Gutierrez-Puente, Y., de Oca-Luna, R. M., Valdes-Flores, J., and Saucedo-Cardenas, O. (2009) Production of biologically active human lymphotactin (XCL1) by Lactococcus lactis. Biotechnol. Lett. 31, 215–220.
van Asseldonk, M., de Vos, W. M., and Simons, G. (1993) Functional analysis of the Lactococcus lactis usp45 secretion signal in the secretion of a homologous proteinase and a heterologous alpha-amylase. Mol. Gen. Genet. 240, 428–434.
Hanniffy, S. B., Carter, A. T., Hitchin, E., and Wells, J. M. (2007) Mucosal delivery of a pneumococcal vaccine using Lactococcus lactis affords protection against respiratory infection. J. Infect. Dis. 195, 185–193.
Bermudez-Humaran, L. G., Cortes-Perez, N. G., Le Loir, Y., Alcocer-Gonzalez, J. M., Tamez-Guerra, R. S., de Oca-Luna, R. M., and Langella, P. (2004) An inducible surface presentation system improves cellular immunity against human papillomavirus type 16 E7 antigen in mice after nasal administration with recombinant lactococci. J. Med. Microbiol. 53, 427–433.
Villatoro-Hernandez, J., Arce-Mendoza, A. Y., Rosas-Taraco, A. G., Esparza-Gonzalez, S. C., Guzman-Lopez, S., Elizondo-Omana, R. E., Chavez-Reyes, A., Saucedo-Cardenas, O., and de Oca Luna, R. M. (2009) Murine interferon-gamma inducible protein-10 (IP-10) secreted by Lactococcus lactis chemo-attracts human CD3+ lymphocytes. Biotechnol. Lett. 31, 1795–1800.
van de Guchte, M., van der Vossen, J. M., Kok, J., and Venema, G. (1989) Construction of a lactococcal expression vector: expression of hen egg white lysozyme in Lactococcus lactis subsp. lactis. Appl Environ Microbiol 55, 224–228.
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Villatoro-Hernández, J., Kuipers, O.P., Saucedo-Cárdenas, O., Montes-de-Oca-Luna, R. (2012). Heterologous Protein Expression by Lactococcus lactis . In: Lorence, A. (eds) Recombinant Gene Expression. Methods in Molecular Biology, vol 824. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-433-9_8
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DOI: https://doi.org/10.1007/978-1-61779-433-9_8
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