Whole-Genome Sequencing of Gliadin Degrading Genes Bacillus amyloliquefaciens Strain via Nanopore Technology

Background: Gluten, a major protein found in wheat, rye, and barley, plays a crucial role in dough formation. Among its components, α-, β-, and γ-gliadins are resistant to enzymatic hydrolysis and can accumulate in the human digestive tract, potentially triggering celiac disease (CD) or nonceliac gluten sensitivity in genetically predisposed individuals. Objective: This study aimed to identify and characterize a bacterial strain capable of degrading gliadin via whole-genome sequencing and comparative genomic analysis. Methods: A gliadin-degrading Bacillus amyloliquefaciens strain , designated SNU-TC2, was isolated on the basis of its ability to form a clear zone on gliadin-containing agar. Whole-genome sequencing was performed viaOxford Nanopore Technology, followed by genome assembly and annotation. Taxonomic classification and genomic comparisons were conducted viaFastANI, core gene phylogenetic, and SNP-based analyses. Results: SNU-TC2 showed >97% nucleotide identity to reference B. amyloliquefaciens strains, with the highest similarity (98.66%) to RD7-7. However, SNP-based comparisonsrevealed 30,736 variants, including several functionally significant mutations. These findings suggest that, while taxonomically similar, SNU-TC2 may harbor distinct genetic features relevant to gliadin degradation. Conclusions: SNU-TC2 represents a promising candidate for further functional validation studies aimed at gluten degradation. Its genomic distinctiveness within the B. amyloliquefaciens strain complex warrants continued investigation for potential application in celiac disease management .