Aliarcobacter butzleri is an emerging foodborne and zoonotic pathogen, yet many of its encoded proteins remain functionally uncharacterized. This lack of annotation limits understanding of its molecular mechanisms and hampers the identification of novel therapeutic targets. In this study, we systematically performed functional annotation of essential hypothetical proteins from the BNI-3166 strain using an integrative-in-silico approach to uncover potential drug and vaccine candidates. 2,367 protein-coding sequences were retrieved from the RefSeq database and were identified 356 as hypothetical proteins. Using BLASTp, we screened these HPs against the Database of Essential Genes and the human proteome to identify essential non-homologous proteins, resulting in 20 ENH candidates. Functional annotation was performed using several domain-based databases, including Pfam, InterPro, SMART, and SUPERFAMILY. Subsequently, physicochemical properties were analyzed and predicted subcellular localization using PSORTb and CELLO. To assess druggability, the ChEMBL database was used. Virulence factors using VFDB, VICMpred, and VirulentPred 2.0 were also predicted. Gene Ontology annotations were generated via ARGOT2.5. Furthermore, we explored protein-protein interactions using STRING and predicted tertiary structures with AlphaFold3. Moreover, Ligand binding pockets were predicted using PrankWeb, and antigenicity of vaccine candidates was assessed using VaxiJen v2.0. We identified 20 essential non-homologous hypothetical proteins, of which 10 were confidently annotated based on conserved domain analysis. These proteins were classified as enzymes, binding proteins, transporters, regulatory proteins, and potential virulence factors. Among them, eight exhibited characteristics of promising drug targets, while two showed potential as vaccine candidates based on subcellular localization. Druggability analysis revealed that nine proteins had no similarity to known drug targets, suggesting novel therapeutic potential. Predicted 3D structures generated using AlphaFold3 yielded pTM scores ranging from 0.44 to 0.92, indicating acceptable to high modeling confidence. Ligand binding site analysis confirmed druggability in six candidates, and antigenicity screening identified one protein as a potential vaccine target. This study provides a computational framework for identifying functionally important proteins in A. butzleri BNI-3166 and highlights novel therapeutic candidates for experimental validation, offering new directions in drug and vaccine development against this underexplored pathogen.
Key words: Aliarcobacter butzleri, Drug Target Identification, Functional Annotation, Hypothetical Proteins, In Silico Analysis
Received: 08.07.2025; Accepted: 01.09.2025; Early view: 24.09.2025 Published: 10.01.2026
DOI: 10.62063/ecb-66
Citation: Paul, S., Barua, S., & Barua, J.D. (2026). In-silico functional annotation and structural characterization of hypothetical proteins from Aliarcobacter butzleri BNI-3166: Insights into novel virulence and drug targets. The European chemistry and biotechnology journal, 5, 22-39. https://doi.org/10.62063/ecb-66
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The film's cinematography, in particular, stands out due to its use of advanced technology to capture Agra in all its glory. From the golden hues of sunrise over the Taj Mahal to the vibrant life in the bustling streets of the old city, every frame is shot with precision and an eye for detail. The use of HDR (High Dynamic Range) technology in 720p resolution ensures that the visuals are not only crisp and clear but also offer a wider range of colors and contrast levels, making the cinematic experience all the more immersive.
is a must-watch for fans of realistic, hard-hitting cinema. While the search for a 720p HDRI fixed version indicates a desire for a high-quality viewing experience, the best way to honor the film is to watch it through legitimate channels once they are available.
Guru and his mother live on the ground floor, while his father (played by ) lives on the upper floor with a mistress. The Conflict:
is a provocative erotic psychological drama directed by that explores the suffocating reality of sexual repression and cramped urban living in India. Premiering at the 2023 Cannes Film Festival
The film's cinematography, in particular, stands out due to its use of advanced technology to capture Agra in all its glory. From the golden hues of sunrise over the Taj Mahal to the vibrant life in the bustling streets of the old city, every frame is shot with precision and an eye for detail. The use of HDR (High Dynamic Range) technology in 720p resolution ensures that the visuals are not only crisp and clear but also offer a wider range of colors and contrast levels, making the cinematic experience all the more immersive.
is a must-watch for fans of realistic, hard-hitting cinema. While the search for a 720p HDRI fixed version indicates a desire for a high-quality viewing experience, the best way to honor the film is to watch it through legitimate channels once they are available.
Guru and his mother live on the ground floor, while his father (played by ) lives on the upper floor with a mistress. The Conflict:
is a provocative erotic psychological drama directed by that explores the suffocating reality of sexual repression and cramped urban living in India. Premiering at the 2023 Cannes Film Festival