Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 18 5 2026 10 05 752 764 Ayu Rahayu Department of Microbiology, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia; Indonesian Biofilm Research Collaboration Center, Yogyakarta, Indonesia Sayyidah Aminy Department of Microbiology, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia; Microbiology Laboratory, Dr. H. Jusuf SK Hospital, Tarakan, North Kalimantan, Indonesia Dian Umaroh Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Tanjungpura, Pontianak, Indonesia Niar Gusnaniar Indonesian Biofilm Research Collaboration Center, Yogyakarta, Indonesia; Research Center for Biomedical Research, National Research and Innovation Agency (BRIN), Cibinong, Bogor, Indonesia Akhirta Atikana Indonesian Biofilm Research Collaboration Center, Yogyakarta, Indonesia; Research Center for Applied Microbiology, National Research and Innovation Agency (BRIN), Cibinong, Bogor, Indonesia Puspita Lisdiyanti Indonesian Biofilm Research Collaboration Center, Yogyakarta, Indonesia; Research Center for Biosystematics and Evolution, National Research and Innovation Agency (BRIN), Cibinong, Bogor, Indonesia Estelle Perraud Laboratoire Ecologie et Biologie des Interactions, Université de Poitiers, Poitiers, France; Laboratory of Parasitology and Mycology, Poitiers University Hospital, Poitiers, France Christine Imbert Laboratory of Parasitology and Mycology, Poitiers University Hospital, Poitiers, France Titik Nuryastuti Department of Microbiology, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia; Indonesian Biofilm Research Collaboration Center, Yogyakarta, Indonesia 2026 01 07 2026 08 06 Background and Objectives: Enterococcus faecium is a major nosocomial pathogen frequently associated with catheter-related infections and biofilm-mediated antimicrobial resistance. This study aimed to investigate the genomic determinants and structural characteristics underlying biofilm formation in a clinical E. faecium isolate recovered from a catheter-associated urinary tract infection (CAUTI). Materials and Methods: A clinical isolate, E. faecium MFKCC-BF.UR33, was subjected to whole-genome sequencing and comprehensive genome annotation to identify biofilm-associated and antimicrobial resistance genes. Phylogenetic analysis was performed to determine its relatedness to reference clinical strains. Structural modeling of selected biofilm-associated proteins was conducted using AlphaFold2 to assess their three-dimensional conformations and potential as therapeutic targets. Results: Whole-genome sequencing revealed a 3.32 Mb genome with a G+C content of 37.35%, comprising 2,321 predicted coding sequences and no detectable plasmids. Genome annotation identified several biofilm-associated genes, including sgrA, acm, fss3, scm, and efbA, as well as antimicrobial resistance genes (antA/B, tetM, tetL, efmA, and liaFSR). Phylogenetic analysis placed the isolate within the E. faecium clade, closely related to other clinical reference strains. Structural modeling demonstrated high-confidence 3D structures for Fss3 and EfbA proteins. Conclusion: The integration of genomic and structural analyses provides insights into the genetic and structural basis of biofilm formation in E. faecium and establishes a foundation for future functional and structure-based investigations. https://ijm.tums.ac.ir/index.php/ijm/article/view/6041 https://ijm.tums.ac.ir/index.php/ijm/article/download/6041/1936