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/).
UltraEdit is a popular text editor software that has been widely used by programmers, developers, and writers for decades. Its powerful features, ease of use, and flexibility have made it a go-to tool for editing and managing text files. However, with the increasing cost of software licenses, many users are looking for alternative solutions, including cracked versions. In this article, we will review UltraEdit-32.v.12.10b.Incl.Keygen crack, a popular cracked version of the software, and provide a comprehensive guide on its features, benefits, and risks.
UltraEdit-32.v.12.10b.Incl.Keygen crack is a popular cracked version of the UltraEdit 12.10b text editor software. While it offers many of the same features as the original software, there are risks associated with using cracked software, including security risks, stability issues, and lack of support. If you need a text editor with advanced features, it is recommended that you consider alternative solutions, such as free and open-source software or purchasing a license.
UltraEdit-32.v.12.10b.Incl.Keygen crack is a cracked version of UltraEdit 12.10b, a text editor software developed by IDM Computer Solutions. The crack includes a keygen, which is a software tool that generates a valid license key for the software, allowing users to bypass the registration process and use the software without purchasing a license.
UltraEdit is a popular text editor software that has been widely used by programmers, developers, and writers for decades. Its powerful features, ease of use, and flexibility have made it a go-to tool for editing and managing text files. However, with the increasing cost of software licenses, many users are looking for alternative solutions, including cracked versions. In this article, we will review UltraEdit-32.v.12.10b.Incl.Keygen crack, a popular cracked version of the software, and provide a comprehensive guide on its features, benefits, and risks.
UltraEdit-32.v.12.10b.Incl.Keygen crack is a popular cracked version of the UltraEdit 12.10b text editor software. While it offers many of the same features as the original software, there are risks associated with using cracked software, including security risks, stability issues, and lack of support. If you need a text editor with advanced features, it is recommended that you consider alternative solutions, such as free and open-source software or purchasing a license.
UltraEdit-32.v.12.10b.Incl.Keygen crack is a cracked version of UltraEdit 12.10b, a text editor software developed by IDM Computer Solutions. The crack includes a keygen, which is a software tool that generates a valid license key for the software, allowing users to bypass the registration process and use the software without purchasing a license.