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
The copyrights of the studies published in The European Chemistry and Biotechnology Journal (EUCHEMBIOJ) belong to their authors
This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0)(https://creativecommons.org/licenses/by-nc/4.0/).
This is not a typical action movie. All Quiet on the Western Front is a slow-burn, emotionally devastating drama that will stay with you long after the credits roll.
Please be aware of copyright laws. Downloading copyrighted content from unofficial sources may be illegal in your country. This guide is for educational purposes for users who own a legal copy of the film and wish to add Indonesian subtitles to it.
, meaning the only legal and official way to download or stream it with Indonesian subtitles is through the Netflix platform Where to Watch Legally Netflix Indonesia Subtitle Options : The platform provides official Indonesian subtitles
: All Quiet on the Western Front (German: Im Westen nichts Neues ).
"All Quiet on the Western Front" 2022 is a powerful and thought-provoking film that is definitely worth watching. By downloading the film with subtitle Indonesia, Indonesian audiences can enjoy the movie in their native language and appreciate the story and characters more easily. However, be sure to take the necessary precautions when downloading the film, and respect the creators and intellectual property rights.
Bagi Anda yang mencarinya, baik melalui streaming resmi di Netflix ataupun jalur unduhan, pastikan Anda dapat menikmatinya dengan subtitle Bahasa Indonesia. Selamat menonton dan bersiaplah untuk perjalanan emosional yang tak akan terlupakan!
: Saat memutar film yang diunduh, masuk ke menu Audio & Subtitles lalu pilih "Indonesian". Waspada Risiko Situs Unduhan Ilegal
Movie Review: All Quiet on the Western Front (2022) - Inkwell
This is not a typical action movie. All Quiet on the Western Front is a slow-burn, emotionally devastating drama that will stay with you long after the credits roll.
Please be aware of copyright laws. Downloading copyrighted content from unofficial sources may be illegal in your country. This guide is for educational purposes for users who own a legal copy of the film and wish to add Indonesian subtitles to it.
, meaning the only legal and official way to download or stream it with Indonesian subtitles is through the Netflix platform Where to Watch Legally Netflix Indonesia Subtitle Options : The platform provides official Indonesian subtitles
: All Quiet on the Western Front (German: Im Westen nichts Neues ).
"All Quiet on the Western Front" 2022 is a powerful and thought-provoking film that is definitely worth watching. By downloading the film with subtitle Indonesia, Indonesian audiences can enjoy the movie in their native language and appreciate the story and characters more easily. However, be sure to take the necessary precautions when downloading the film, and respect the creators and intellectual property rights.
Bagi Anda yang mencarinya, baik melalui streaming resmi di Netflix ataupun jalur unduhan, pastikan Anda dapat menikmatinya dengan subtitle Bahasa Indonesia. Selamat menonton dan bersiaplah untuk perjalanan emosional yang tak akan terlupakan!
: Saat memutar film yang diunduh, masuk ke menu Audio & Subtitles lalu pilih "Indonesian". Waspada Risiko Situs Unduhan Ilegal
Movie Review: All Quiet on the Western Front (2022) - Inkwell