Background <p>Celiac disease (CeD), a multifactorial disorder, develops when gluten, the toxic environmental inducer, interacts with CeD susceptibility genetic markers, resulting in a chronic enteropathy. Several extra-intestinal complications may also arise in cases of delayed management. There persists a growing demand to develop non-dietary adjuvant therapeutic options that can help relieve symptoms and improve patients’ quality of life.</p> Aim <p>The present study conducted a bioinformatic approach to look into the potential of using <i>Lactobacillus rhamnosus</i>, a well-established probiotic, as gluten-digesting bacteria and provide the basis for future therapeutic developments.</p> Methods <p>Complete genome assemblies of forty-nine <i>L. rhamnosus</i> strains were subjected to annotation using RAST and a pan genome analysis with BPGA. Genes for peptidases were identified using BlastKOALA and Prokka, followed by domain analysis using the NCBI-CD search tool to screen for gluten-digesting activity.</p> Results <p>Genome annotation of all the strains under study highlighted the presence of sixty-one peptidases in <i>L. rhamnosus</i>. Domain analysis further revealed that nine of these peptidases, including aminopeptidase N, neutral endopeptidase, oligoendopeptidase F, dipeptidyl-peptidase 5, proline iminopeptidase, Xaa-Pro dipeptidyl-peptidase, aminopeptidase C, aminopeptidase E, and PII-type proteinase, shared domains with already established gluten-digesting enzymes, suggesting their potential role in degrading toxic gliadin peptides.</p> Conclusion <p>The current in silico analysis indicates that this well-known probiotic species, in addition to showcasing a plethora of beneficial properties, may also hold great potential in terms of reducing gluten toxicity. With further studies, <i>L. rhamnosus</i> can prove to be a promising candidate in CeD treatment and management.</p> Graphical abstract <p></p>