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Mechanism of degradation of immunogenic gluten epitopes from Triticum turgidum L. var. durum by sourdough lactobacilli and fungal proteases.
Appl Environ Microbiol. 2010 Jan; 76(2):508-18.AE

Abstract

As shown by R5 antibody-based sandwich and competitive enzyme-linked immunosorbent assay (ELISA), selected sourdough lactobacilli, in combination with fungal proteases, hydrolyzed gluten (72 h at 37 degrees C) of various cultivars of Triticum turgidum L. var. durum to less than 20 ppm. Complementary electrophoretic, chromatography, and mass spectrometry techniques were used to characterize the gluten and epitope hydrolysis. Nine peptidases were partially purified from the pooled cytoplasmic extract of the sourdough lactobacilli and used to hydrolyze the 33-mer epitope, the most immunogenic peptide generated during digestion of Triticum species. At least three peptidases (general aminopeptidase type N [PepN], X-prolyl dipeptidyl aminopeptidase [PepX], and endopeptidase PepO) were necessary to detoxify the 33-mer without generation of related immunogenic epitopes. After 14 h of incubation, the combination of all or at least six different peptidases totally hydrolyzed the 33-mer (200 mM) into free amino acids. The same results were found for other immunogenic epitopes, such as fragments 57-68 of alpha 9-gliadin, 62-75 of A-gliadin, and 134-153 of gamma-gliadin. When peptidases were used for fermentation of durum wheat semolina, they caused the hydrolysis of gluten to ca. 2 ppm. The in vivo digestion was simulated, and proteins/peptides extracted from pepsin-trypsin (PT) digestion of durum wheat semolina fermented with selected sourdough lactobacilli induced the expression of gamma interferon and interleukin 2 at levels comparable to those of the negative control. Durum wheat semolina fermented with sourdough lactobacilli was freeze-dried and used for making Italian-type pasta. The scores for cooking and sensory properties for this pasta were higher that those of conventional gluten-free pasta.

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Authors+Show Affiliations

De Angelis M
Department of Plant Protection and Applied Microbiology, University of Bari, 70126 Bari, Italy.
Cassone A
No affiliation info available
Rizzello CG
No affiliation info available
Gagliardi F
No affiliation info available
Minervini F
No affiliation info available
Calasso M
No affiliation info available
Di Cagno R
No affiliation info available
Francavilla R
No affiliation info available
Gobbetti M
No affiliation info available

MeSH

AdultAmino Acid SequenceBiopsyCytokinesDuodenumEpitopesFermentationFood HandlingFungiGlutensHumansLactobacillusMolecular Sequence DataPeptide HydrolasesTriticum

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19948868

Citation

De Angelis, Maria, et al. "Mechanism of Degradation of Immunogenic Gluten Epitopes From Triticum Turgidum L. Var. Durum By Sourdough Lactobacilli and Fungal Proteases." Applied and Environmental Microbiology, vol. 76, no. 2, 2010, pp. 508-18.
De Angelis M, Cassone A, Rizzello CG, et al. Mechanism of degradation of immunogenic gluten epitopes from Triticum turgidum L. var. durum by sourdough lactobacilli and fungal proteases. Appl Environ Microbiol. 2010;76(2):508-18.
De Angelis, M., Cassone, A., Rizzello, C. G., Gagliardi, F., Minervini, F., Calasso, M., Di Cagno, R., Francavilla, R., & Gobbetti, M. (2010). Mechanism of degradation of immunogenic gluten epitopes from Triticum turgidum L. var. durum by sourdough lactobacilli and fungal proteases. Applied and Environmental Microbiology, 76(2), 508-18. https://doi.org/10.1128/AEM.01630-09
De Angelis M, et al. Mechanism of Degradation of Immunogenic Gluten Epitopes From Triticum Turgidum L. Var. Durum By Sourdough Lactobacilli and Fungal Proteases. Appl Environ Microbiol. 2010;76(2):508-18. PubMed PMID: 19948868.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Mechanism of degradation of immunogenic gluten epitopes from Triticum turgidum L. var. durum by sourdough lactobacilli and fungal proteases. AU - De Angelis,Maria, AU - Cassone,Angela, AU - Rizzello,Carlo G, AU - Gagliardi,Francesca, AU - Minervini,Fabio, AU - Calasso,Maria, AU - Di Cagno,Raffaella, AU - Francavilla,Ruggero, AU - Gobbetti,Marco, Y1 - 2009/11/30/ PY - 2009/12/2/entrez PY - 2009/12/2/pubmed PY - 2010/3/10/medline SP - 508 EP - 18 JF - Applied and environmental microbiology JO - Appl Environ Microbiol VL - 76 IS - 2 N2 - As shown by R5 antibody-based sandwich and competitive enzyme-linked immunosorbent assay (ELISA), selected sourdough lactobacilli, in combination with fungal proteases, hydrolyzed gluten (72 h at 37 degrees C) of various cultivars of Triticum turgidum L. var. durum to less than 20 ppm. Complementary electrophoretic, chromatography, and mass spectrometry techniques were used to characterize the gluten and epitope hydrolysis. Nine peptidases were partially purified from the pooled cytoplasmic extract of the sourdough lactobacilli and used to hydrolyze the 33-mer epitope, the most immunogenic peptide generated during digestion of Triticum species. At least three peptidases (general aminopeptidase type N [PepN], X-prolyl dipeptidyl aminopeptidase [PepX], and endopeptidase PepO) were necessary to detoxify the 33-mer without generation of related immunogenic epitopes. After 14 h of incubation, the combination of all or at least six different peptidases totally hydrolyzed the 33-mer (200 mM) into free amino acids. The same results were found for other immunogenic epitopes, such as fragments 57-68 of alpha 9-gliadin, 62-75 of A-gliadin, and 134-153 of gamma-gliadin. When peptidases were used for fermentation of durum wheat semolina, they caused the hydrolysis of gluten to ca. 2 ppm. The in vivo digestion was simulated, and proteins/peptides extracted from pepsin-trypsin (PT) digestion of durum wheat semolina fermented with selected sourdough lactobacilli induced the expression of gamma interferon and interleukin 2 at levels comparable to those of the negative control. Durum wheat semolina fermented with sourdough lactobacilli was freeze-dried and used for making Italian-type pasta. The scores for cooking and sensory properties for this pasta were higher that those of conventional gluten-free pasta. SN - 1098-5336 UR - https://www.unboundmedicine.com/medline/citation/19948868/Mechanism_of_degradation_of_immunogenic_gluten_epitopes_from_Triticum_turgidum_L__var__durum_by_sourdough_lactobacilli_and_fungal_proteases_ L2 - https://journals.asm.org/doi/10.1128/AEM.01630-09?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -