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Applied Microbiology and Biotechnology

, Volume 98, Issue 15, pp 6689–6700 | Cite as

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

  • Patricia Alvarez-Sieiro
  • Maria Cruz Martin
  • Begoña Redruello
  • Beatriz del Rio
  • Victor Ladero
  • Brad A. Palanski
  • Chaitan Khosla
  • Maria Fernandez
  • Miguel A. Alvarez
Applied genetics and molecular biotechnology

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.

Keywords

Celiac disease Gluten Prolyl endopeptidase Myxococcus xanthus Heterologous expression Lactobacillus casei 

Notes

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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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Patricia Alvarez-Sieiro
    • 1
  • Maria Cruz Martin
    • 1
  • Begoña Redruello
    • 1
  • Beatriz del Rio
    • 1
  • Victor Ladero
    • 1
  • Brad A. Palanski
    • 2
    • 3
  • Chaitan Khosla
    • 2
    • 3
  • Maria Fernandez
    • 1
  • Miguel A. Alvarez
    • 1
  1. 1.Instituto de Productos Lácteos de Asturias (IPLA-CSIC)VillaviciosaSpain
  2. 2.Department of ChemistryStanford UniversityStanfordUSA
  3. 3.Department of Chemical EngineeringStanford UniversityStanfordUSA

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