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Generation of food-grade recombinant Lactobacillus casei delivering Myxococcus xanthus prolyl endopeptidase

  • Applied genetics and molecular biotechnology
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Abstract

Prolyl endopeptidases (PEP) (EC 3.4.21.26), a family of serine proteases with the ability to hydrolyze the peptide bond on the carboxyl side of an internal proline residue, are able to degrade immunotoxic peptides responsible for celiac disease (CD), such as a 33-residue gluten peptide (33-mer). Oral administration of PEP has been suggested as a potential therapeutic approach for CD, although delivery of the enzyme to the small intestine requires intrinsic gastric stability or advanced formulation technologies. We have engineered two food-grade Lactobacillus casei strains to deliver PEP in an in vitro model of small intestine environment. One strain secretes PEP into the extracellular medium, whereas the other retains PEP in the intracellular environment. The strain that secretes PEP into the extracellular medium is the most effective to degrade the 33-mer and is resistant to simulated gastrointestinal stress. Our results suggest that in the future, after more studies and clinical trials, an engineered food-grade Lactobacillus strain may be useful as a vector for in situ production of PEP in the upper small intestine of CD patients.

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Acknowledgments

The authors are grateful to Isabel Cuesta and Jorge R. Álvarez-Buylla for their technical assistance. This research was supported by project 201370E094 from the CSIC. P.A. is the recipient of a fellowship from FICYT (BP09093) and B.D.R. is a beneficiary of a JAE-DOC contract (CSIC). C.K. is supported by a grant from the NIH (R01 DK 063158).

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Authors and Affiliations

  1. Instituto de Productos Lácteos de Asturias (IPLA-CSIC), 33300, Villaviciosa, Asturias, Spain

    Patricia Alvarez-Sieiro, Maria Cruz Martin, Begoña Redruello, Beatriz del Rio, Victor Ladero, Maria Fernandez & Miguel A. Alvarez

  2. Department of Chemistry, Stanford University, Stanford, CA, 94305, USA

    Brad A. Palanski & Chaitan Khosla

  3. Department of Chemical Engineering, Stanford University, Stanford, CA, 94305, USA

    Brad A. Palanski & Chaitan Khosla

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  1. Patricia Alvarez-Sieiro
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  2. Maria Cruz Martin
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  3. Begoña Redruello
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  4. Beatriz del Rio
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  8. Maria Fernandez
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  9. Miguel A. Alvarez
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Correspondence to Miguel A. Alvarez.

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Alvarez-Sieiro, P., Martin, M.C., Redruello, B. et al. Generation of food-grade recombinant Lactobacillus casei delivering Myxococcus xanthus prolyl endopeptidase. Appl Microbiol Biotechnol 98, 6689–6700 (2014). https://doi.org/10.1007/s00253-014-5730-7

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