Microbial Peptidases: Key Players in Reducing Gluten Immunogenicity Through Peptide Degradation
Source abstract
Abstract
Gluten-related disorders, including celiac disease (CeD) and non-celiac gluten sensitivity (NCGS), are triggered by the immune response to gluten peptides that resist complete digestion by human gastrointestinal enzymes. Microbial peptidases have emerged as promising biocatalysts capable of degrading these immunogenic peptides, offering potential therapeutic and industrial applications. This review explores the role of microbial peptidases in gluten degradation, highlighting key enzyme families, their mechanisms of action, and their effectiveness in reducing gluten immunogenicity. Additionally, we discuss advances in enzymatic therapy, food processing applications, and the challenges associated with optimizing microbial enzymes for safe and efficient gluten detoxification. Understanding the potential of microbial peptidases in mitigating gluten-related disorders paves the way for novel dietary and therapeutic strategies.
Source: crossref · Quality: baseline
Research context
OpenAlex topics: Celiac Disease Research and Management · Peptidase Inhibition and Analysis · Eosinophilic Esophagitis
SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.
Affiliations
Area Biochemistry, Department of Molecular Biology, University of León, 24071 León, Spain
Instituto de Biología Molecular, Genómica y Proteómica (INBIOMIC), University of León, Campus de Vegazana, 24071 León, Spain
Funding
The University of León · Spanish Ministerio de Ciencia e Innovación · Junta de Castilla y León
References (99)
Show source-provided references
1. Scherf, K.A. (2025). Gluten and Wheat-Related Disorders. Handbook of Food Allergen Detection and Control, Elsevier.10.1016/B978-0-12-821733-7.00008-2 ↗
2. Catassi, G., Marmo, C., Gasbarrini, A., and Riccioni, M.E. (2024). Role of Device-Assisted Enteroscopy in Crohn’s Disease. J. Clin. Med., 13.10.3390/jcm13133919 ↗
3. Gluten-Dependent Activation of CD4+ T Cells by MHC Class II–Expressing Epithelium10.1053/j.gastro.2024.07.008 ↗
4. San-Martin, M.I., Chamizo-Ampudia, A., Sanchiz, Á., Ferrero, M.Á., Martínez-Blanco, H., Rodríguez-Aparicio, L.B., and Navasa, N. (2025). Microbiome Markers in Gastrointestinal Disorders: Inflammatory Bowel Disease, Colorectal Cancer, and Celiac Disease. Int. J. Mol. Sci., 26.10.3390/ijms26104818 ↗
5. Caio, G., Volta, U., Sapone, A., Leffler, D.A., De Giorgio, R., Catassi, C., and Fasano, A. (2019). Celiac Disease: A Comprehensive Current Review. BMC Med., 17.10.1186/s12916-019-1380-z ↗
6. Celiac Disease in Asia beyond the Middle East and Indian Subcontinent: Epidemiological Burden and Diagnostic Barriers10.3748/wjg.v27.i19.2251 ↗
7. Catassi, C., Alaedini, A., Bojarski, C., Bonaz, B., Bouma, G., Carroccio, A., Castillejo, G., De Magistris, L., Dieterich, W., and Di Liberto, D. (2017). The Overlapping Area of Non-Celiac Gluten Sensitivity (NCGS) and Wheat-Sensitive Irritable Bowel Syndrome (IBS): An Update. Nutrients, 9.10.3390/nu9111268 ↗
8. Maintaining a Balanced Diet While Gluten-Free: Treatment Options10.1007/s11938-020-00319-6 ↗
9. Wieser, H., Segura, V., Ruiz-Carnicer, Á., Sousa, C., and Comino, I. (2021). Food Safety and Cross-Contamination of Gluten-Free Products: A Narrative Review. Nutrients, 13.10.3390/nu13072244 ↗
10. Liu, S., El Khoury, D., and Joye, I.J. (2023). Gluten-Free Product Recalls and Their Impact on Consumer Trust. Nutrients, 15.10.3390/nu15194170 ↗
11. Lessons from Low-Gluten Challenge Studies in Celiac Disease: Is It Time to Reconsider the Gluten Threshold in Gluten-Free Food?10.1016/j.dld.2025.01.202 ↗
12. Wheat Breeding, Fertilizers, and Pesticides: Do They Contribute to the Increasing Immunogenic Properties of Modern Wheat?10.3390/gidisord3040023 ↗
13. Ali, A., and AlHussaini, K.I. (2024). Pesticides: Unintended Impact on the Hidden World of Gut Microbiota. Metabolites, 14.10.3390/metabo14030155 ↗
14. Sawant, S.S., Park, H.-Y., Sim, E.-Y., Kim, H.-S., and Choi, H.-S. (2025). Microbial Fermentation in Food: Impact on Functional Properties and Nutritional Enhancement—A Review of Recent Developments. Fermentation, 11.10.3390/fermentation11010015 ↗
15. Current Approaches to Diagnosis and Treatment of Celiac Disease: An Evolving Spectrum10.1053/gast.2001.22123 ↗
16. Detoxification Mechanism and the Impact of Transamidation-Modified Gliadin on Celiac-Based Gluten Sensitivity: The Potential of Unlocking Gluten Tolerance in Functional Food10.1021/acs.jafc.4c10318 ↗
17. How to Reduce Gluten in Foods: A Critical Review of Patents10.1111/ijfs.17550 ↗
18. Mamone, G., Di Stasio, L., Vitale, S., Picascia, S., and Gianfrani, C. (2023). Analytical and Functional Approaches to Assess the Immunogenicity of Gluten Proteins. Front. Nutr., 9.10.3389/fnut.2022.1049623 ↗
19. Hydrolytic Enzymes and Their Directly and Indirectly Effects on Gluten and Dough Properties: An Extensive Review10.1002/fsn3.2344 ↗
20. Scherf, K.A., Catassi, C., Chirdo, F., Ciclitira, P.J., Feighery, C., Gianfrani, C., Koning, F., Lundin, K.E.A., Schuppan, D., and Smulders, M.J.M. (2020). Recent Progress and Recommendations on Celiac Disease From the Working Group on Prolamin Analysis and Toxicity. Front. Nutr., 7.10.3389/fnut.2020.00029 ↗
21. A Review of Gluten Detoxification in Wheat for Food Applications: Approaches, Mechanisms, and Implications10.1080/10408398.2024.2326618 ↗
22. Effects of Thermal Treatment on the Physicochemical and Structural Properties of Wheat Gluten Proteins: Insights from Gluten, Glutenin, and Gliadin Fractions10.1111/ijfs.16953 ↗
23. The Effect of Baking Time and Temperature on Gluten Protein Structure and Celiac Peptide Digestibility10.1016/j.foodres.2020.109988 ↗
24. A High-Affinity Human TCR-like Antibody Detects Celiac Disease Gluten Peptide–MHC Complexes and Inhibits T Cell Activation10.1126/sciimmunol.abg4925 ↗
25. Gluten-Related Disorders: Celiac Disease, Wheat Allergy, and Nonceliac Gluten Sensitivity10.1080/10408398.2019.1651689 ↗
26. A Critical Review on the Gluten-Induced Enteropathy/Celiac Disease: Gluten-Targeted Dietary and Non-Dietary Therapeutic Approaches10.1080/87559129.2023.2202405 ↗
27. Baharin, A., Ting, T.-Y., and Goh, H.-H. (2022). Post-Proline Cleaving Enzymes (PPCEs): Classification, Structure, Molecular Properties, and Applications. Plants, 11.10.20944/preprints202204.0097.v1 ↗
28. Society for the Study of Celiac Disease Position Statement on Gaps and Opportunities in Coeliac Disease10.1038/s41575-021-00511-8 ↗
29. de Lourdes Moreno, M., Segura, V., Ruiz-Carnicer, Á., Nájar, A.M., Comino, I., and Sousa, C. (2021). Oral Enzyme Strategy in Celiac Disease. Biotechnological Strategies for the Treatment of Gluten Intolerance, Elsevier.10.1016/B978-0-12-821594-4.00005-0 ↗
30. Cristofori, F., Francavilla, R., Capobianco, D., Dargenio, V.N., Filardo, S., and Mastromarino, P. (2020). Bacterial-Based Strategies to Hydrolyze Gluten Peptides and Protect Intestinal Mucosa. Front. Immunol., 11.10.3389/fimmu.2020.567801 ↗
31. Wheat Gluten: A Functional Protein Still Challenging to Replace in Gluten-free Cereal-based Foods10.1002/cche.10624 ↗
32. Stanciu, D., Staykov, H., Dragomanova, S., Tancheva, L., Pop, R., Ielciu, I., and Crișan, G. (2024). Gluten Unraveled: Latest Insights on Terminology, Diagnosis, Pathophysiology, Dietary Strategies, and Intestinal Microbiota Modulations—A Decade in Review. Nutrients, 16.10.3390/nu16213636 ↗
33. Celiac Disease and Immunogenic Wheat Gluten Peptides and the Association of Gliadin Peptides with HLA DQ2 and HLA DQ810.1080/87559129.2021.1907755 ↗
34. Rani, M., Sogi, D.S., and Gill, B.S. (2021). Characterization of Gliadin, Secalin and Hordein Fractions Using Analytical Techniques. Sci. Rep., 11.10.1038/s41598-021-02099-0 ↗
35. Voisine, J., and Abadie, V. (2021). Interplay Between Gluten, HLA, Innate and Adaptive Immunity Orchestrates the Development of Coeliac Disease. Front. Immunol., 12.10.3389/fimmu.2021.674313 ↗
36. Lindstad, C.B., Dewan, A.E., Stamnaes, J., Sollid, L.M., and du Pré, M.F. (2021). TG2-Gluten Complexes as Antigens for Gluten-Specific and Transglutaminase-2 Specific B Cells in Celiac Disease. PLoS ONE, 16.10.1371/journal.pone.0259082 ↗
37. Oligomerization of 33-mer Gliadin Peptides: Supramolecular Assemblies in Celiac Disease10.1002/cmdc.202400789 ↗
38. Dunne, M.R., Byrne, G., Chirdo, F.G., and Feighery, C. (2020). Coeliac Disease Pathogenesis: The Uncertainties of a Well-Known Immune Mediated Disorder. Front. Immunol., 11.10.3389/fimmu.2020.01374 ↗
39. Cardo, A., Churruca, I., Lasa, A., Navarro, V., Vázquez-Polo, M., Perez-Junkera, G., and Larretxi, I. (2021). Nutritional Imbalances in Adult Celiac Patients Following a Gluten-Free Diet. Nutrients, 13.10.3390/nu13082877 ↗
40. IL-15 Functions as a Danger Signal to Regulate Tissue-Resident T Cells and Tissue Destruction10.1038/nri3919 ↗
41. Gluten-Degrading Bacteria: Availability and Applications10.1007/s00253-021-11263-5 ↗
42. Structural Basis for Gluten Intolerance in Celiac Sprue10.1126/science.1074129 ↗
43. Intestinal Digestive Resistance of Immunodominant Gliadin Peptides10.1152/ajpgi.00136.2002 ↗
44. Nutrition Assessment and Management in Celiac Disease
45. Wei, G., Helmerhorst, E.J., Darwish, G., Blumenkranz, G., and Schuppan, D. (2020). Gluten Degrading Enzymes for Treatment of Celiac Disease. Nutrients, 12.10.3390/nu12072095 ↗
46. Freitas, D., Gómez-Mascaraque, L.G., and Brodkorb, A. (2022). Digestion of Protein and Toxic Gluten Peptides in Wheat Bread, Pasta and Cereal and the Effect of a Supplemental Enzyme Mix. Front. Nutr., 9.10.3389/fnut.2022.986272 ↗
47. Segura, V., Ruiz-Carnicer, Á., Sousa, C., and Moreno, M.D.L. (2021). New Insights into Non-Dietary Treatment in Celiac Disease: Emerging Therapeutic Options. Nutrients, 13.10.3390/nu13072146 ↗
48. Dunaevsky, Y.E., Tereshchenkova, V.F., Belozersky, M.A., Filippova, I.Y., Oppert, B., and Elpidina, E.N. (2021). Effective Degradation of Gluten and Its Fragments by Gluten-Specific Peptidases: A Review on Application for the Treatment of Patients with Gluten Sensitivity. Pharmaceutics, 13.10.3390/pharmaceutics13101603 ↗
49. New Coeliac Disease Treatments and Their Complications10.1016/j.gastrohep.2017.12.002 ↗
50. Fabry Disease: Current and Novel Therapeutic Strategies. A Narrative Review10.2174/1570159X20666220601124117 ↗
51. Industrial Enzyme Production in Bangladesh: Current Landscape, Scope, and Challenges10.3329/ajmbr.v9i4.69395 ↗
52. Digestive Enzyme Supplementation in Gastrointestinal Diseases10.2174/138920021702160114150137 ↗
53. Biotransformation of Wheat Proteins with Lactobacillus Sp. Following High Hydrostatic Pressure Pre-Treatment to Reduce Gluten Immunoreactivity10.1016/j.jcs.2023.103787 ↗
54. Fermentation, Purification, Formulation, and Pharmacological Evaluation of a Prolyl Endopeptidase from Myxococcus Xanthus: Implications for Celiac Sprue Therapy10.1002/bit.20643 ↗
55. Prolyl Endopeptidase FromSphingomonas Capsulata:Isolation and Characterization of the Enzyme and Nucleotide Sequence of the Gene10.1006/abbi.1998.0836 ↗
56. Cloning of Proline-Specific Endopeptidase Gene from Flavobacterium Meningosepticum: Expression in Escherichia Coli and Purification of the Heterologous Protein10.1007/BF00170434 ↗
57. Unconventional Serine Proteases: Variations on the Catalytic Ser/His/Asp Triad Configuration10.1110/ps.035436.108 ↗
58. Structural and Time-resolved Mechanistic Investigations of Protein Hydrolysis by the Acidic Proline-specific Endoprotease from Aspergillus Niger10.1002/pro.4856 ↗
59. Xiao, B., Zhang, C., Zhou, J., Wang, S., Meng, H., Wu, M., Zheng, Y., and Yu, R. (2023). Design of SC PEP with Enhanced Stability against Pepsin Digestion and Increased Activity by Machine Learning and Structural Parameters Modeling. Int. J. Biol. Macromol., 250.10.1016/j.ijbiomac.2023.125933 ↗
60. Effect of Aspergillus Niger Prolyl Endopeptidase in Patients with Celiac Disease on a Long-Term Gluten-Free Diet10.3748/wjg.v30.i11.1545 ↗
61. König, J., Holster, S., Bruins, M.J., and Brummer, R.J. (2017). Randomized Clinical Trial: Effective Gluten Degradation by Aspergillus Niger-Derived Enzyme in a Complex Meal Setting. Sci. Rep., 7.10.1038/s41598-017-13587-7 ↗
62. Industrial Scope of Cysteine Protease and Its Anti-Gluten Property: A Review10.1016/j.fnutr.2025.100006 ↗
63. Effect of Barley Endoprotease EP-B2 on Gluten Digestion in the Intact Rat10.1124/jpet.106.104315 ↗
64. Heterologous Expression, Purification, Refolding, and Structural-Functional Characterization of EP-B2, a Self-Activating Barley Cysteine Endoprotease10.1016/j.chembiol.2006.04.008 ↗
65. Papain-like Cysteine Proteinase Zone (PCP-Zone) and PCP Structural Catalytic Core (PCP-SCC) of Enzymes with Cysteine Proteinase Fold10.1016/j.ijbiomac.2020.10.022 ↗
66. Bethune, M.T., Ribka, E., Khosla, C., and Sestak, K. (2008). Transepithelial Transport and Enzymatic Detoxification of Gluten in Gluten-Sensitive Rhesus Macaques. PLoS ONE, 3.10.1371/journal.pone.0001857 ↗
67. A Scaleable Manufacturing Process for Pro-EP-B2, a Cysteine Protease from Barley Indicated for Celiac Sprue10.1002/bit.21423 ↗
68. Yoosuf, S., and Makharia, G.K. (2019). Evolving Therapy for Celiac Disease. Front. Pediatr., 7.10.3389/fped.2019.00193 ↗
69. Darwish, G., Helmerhorst, E.J., Schuppan, D., Oppenheim, F.G., and Wei, G. (2019). Pharmaceutically Modified Subtilisins Withstand Acidic Conditions and Effectively Degrade Gluten in Vivo. Sci. Rep., 9.10.1038/s41598-019-43837-9 ↗
70. Wei, G., Darwish, G., Oppenheim, F.G., Schuppan, D., and Helmerhorst, E.J. (2020). Commensal Bacterium Rothia Aeria Degrades and Detoxifies Gluten via a Highly Effective Subtilisin Enzyme. Nutrients, 12.10.3390/nu12123724 ↗
71. Identification of Food-Grade Subtilisins as Gluten-Degrading Enzymes to Treat Celiac Disease10.1152/ajpgi.00185.2016 ↗
72. Zamakhchari, M., Wei, G., Dewhirst, F., Lee, J., Schuppan, D., Oppenheim, F.G., and Helmerhorst, E.J. (2011). Identification of Rothia Bacteria as Gluten-Degrading Natural Colonizers of the Upper Gastro-Intestinal Tract. PLoS ONE, 6.10.1371/journal.pone.0024455 ↗
73. Salivary Gluten Degradation and Oral Microbial Profiles in Healthy Individuals and Celiac Disease Patients10.1128/AEM.03330-16 ↗
74. Microbial-Derived Peptidases Are Altered in Celiac Disease, Non-Celiac Gluten Sensitivity, and Functional Dyspepsia: A Systematic Review and Re-Analysis of the Duodenal Microbiome10.1080/19490976.2025.2500063 ↗
75. Kriaa, A., Jablaoui, A., Rhimi, S., Soussou, S., Mkaouar, H., Mariaule, V., Gruba, N., Gargouri, A., Maguin, E., and Lesner, A. (2021). SP-1, a Serine Protease from the Gut Microbiota, Influences Colitis and Drives Intestinal Dysbiosis in Mice. Cells, 10.10.3390/cells10102658 ↗
76. Maciejewska, M., Dąbrowska, A., and Cano-Lamadrid, M. (2025). Sustainable Protein Sources: Functional Analysis of Tenebrio Molitor Hydrolysates and Attitudes of Consumers in Poland and Spain Toward Insect-Based Foods. Foods, 14.10.3390/foods14020333 ↗
77. Dvoryakova, E.A., Klimova, M.A., Simonyan, T.R., Dombrovsky, I.A., Serebryakova, M.V., Tereshchenkova, V.F., Dunaevsky, Y.E., Belozersky, M.A., Filippova, I.Y., and Elpidina, E.N. (2022). Recombinant Cathepsin L of Tribolium Castaneum and Its Potential in the Hydrolysis of Immunogenic Gliadin Peptides. Int. J. Mol. Sci., 23.10.3390/ijms23137001 ↗
78. Selected Probiotic Lactobacilli Have the Capacity To Hydrolyze Gluten Peptides during Simulated Gastrointestinal Digestion10.1128/AEM.00376-17 ↗
79. A Prolyl Endopeptidase from Flammulina Velutipes for the Possible Degradation of Celiac Disease Provoking Toxic Peptides in Cereal Proteins10.1016/j.procbio.2018.07.019 ↗
80. Review Article: Novel Enzyme Therapy Design for Gluten Peptide Digestion Through Exopeptidase Supplementation10.1111/apt.70014 ↗
81. Comparative Biochemical Analysis of Three Bacterial Prolyl Endopeptidases: Implications for Coeliac Sprue10.1042/BJ20040907 ↗
82. Efficient Degradation of Gluten by a Prolyl Endoprotease in a Gastrointestinal Model: Implications for Coeliac Disease10.1136/gut.2006.111609 ↗
83. Helmerhorst, E.J., Zamakhchari, M., Schuppan, D., and Oppenheim, F.G. (2010). Discovery of a Novel and Rich Source of Gluten-Degrading Microbial Enzymes in the Oral Cavity. PLoS ONE, 5.10.1371/journal.pone.0013264 ↗
84. The Cultivable Human Oral Gluten-Degrading Microbiome and Its Potential Implications in Coeliac Disease and Gluten Sensitivity10.1111/1469-0691.12249 ↗
85. Li, J., Li, Y., Zhou, Y., Wang, C., Wu, B., and Wan, J. (2018). Actinomyces and Alimentary Tract Diseases: A Review of Its Biological Functions and Pathology. Biomed. Res. Int., 2018.10.1155/2018/3820215 ↗
86. Commercially Available Glutenases: A Potential Hazard in Coeliac Disease10.1177/1756283X17690991 ↗
87. In Vivo Fluorescence Imaging of Exogenous Enzyme Activity in the Gastrointestinal Tract10.1073/pnas.1100285108 ↗
88. Ma, J., Yan, L., Yang, J., He, Y., and Wu, L. (2024). Effect of Modification Strategies on the Biological Activity of Peptides/Proteins. ChemBioChem, 25.10.1002/cbic.202300481 ↗
89. Chugunov, A.O., Dvoryakova, E.A., Dyuzheva, M.A., Simonyan, T.R., Tereshchenkova, V.F., Filippova, I.Y., Efremov, R.G., and Elpidina, E.N. (2023). Fighting Celiac Disease: Improvement of PH Stability of Cathepsin L In Vitro by Computational Design. Int. J. Mol. Sci., 24.10.3390/ijms241512369 ↗
90. Liu, Y.-Y., Ye, R.-L., and Meng, M. (2023). Specificity Enhancement of Glutenase Bga1903 toward Celiac Disease-Eliciting Pro-Immunogenic Peptides via Active-Site Modification. Int. J. Mol. Sci., 25.10.20944/preprints202311.1601.v1 ↗
91. Glutenase ALV003 Attenuates Gluten-Induced Mucosal Injury in Patients With Celiac Disease10.1053/j.gastro.2014.02.031 ↗
92. The Effects of ALV003 Pre-Digestion of Gluten on Immune Response and Symptoms in Celiac Disease in Vivo10.1016/j.clim.2009.11.001 ↗
93. Latiglutenase Treatment for Celiac Disease: Symptom and Quality of Life Improvement for Seropositive Patients on a Gluten-free Diet10.1002/ygh2.371 ↗
94. Latiglutenase Protects the Mucosa and Attenuates Symptom Severity in Patients With Celiac Disease Exposed to a Gluten Challenge10.1053/j.gastro.2022.07.071 ↗
95. Gluten Degradation, Pharmacokinetics, Safety, and Tolerability of TAK-062, an Engineered Enzyme to Treat Celiac Disease10.1053/j.gastro.2021.03.019 ↗
96. Ehren, J., Morón, B., Martin, E., Bethune, M.T., Gray, G.M., and Khosla, C. (2009). A Food-Grade Enzyme Preparation with Modest Gluten Detoxification Properties. PLoS ONE, 4.10.1371/journal.pone.0006313 ↗
97. Pharmacology, Physiology, and Mechanisms of Action of Dipeptidyl Peptidase-4 Inhibitors10.1210/er.2014-1035 ↗
98. More than Just an Enzyme: Dipeptidyl Peptidase-4 (DPP-4) and Its Association with Diabetic Kidney Remodelling10.1016/j.phrs.2019.104391 ↗
99. Assessing the Association between Dipeptidyl Peptidase-4 Inhibitors Use and Celiac Disease through Drug Adverse Event Reporting10.1177/2040622320904301 ↗