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
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Viewing content through legitimate means ensures that the actors, writers, and production crews are fairly compensated for their work.
Official apps allow for seamless viewing across multiple devices, including smartphones, tablets, and smart TVs, often with the option to download episodes for offline viewing within the app. Digital Safety Best Practices
Ensure that operating systems and security software are up to date to protect against vulnerabilities.
Be cautious of websites that claim to offer free downloads of copyrighted material, as these are often used to distribute malicious software.
Set in the fictional Moordale Secondary School, the show follows Otis Milburn, a socially awkward teenager, and his mother, Jean, who is a professional sex therapist. The series explores a wide range of sensitive topics with humor and heart, making it a favorite for audiences seeking both entertainment and meaningful dialogue. How to Watch Safely and Legally
Always download streaming applications from verified stores like the Apple App Store or Google Play Store.
Using authorized services eliminates the risks associated with third-party sites, such as malware, phishing, or intrusive advertisements.
By choosing legitimate streaming options, viewers can enjoy the journey of Otis, Maeve, and the rest of the Moordale community while ensuring a safe and high-quality experience.
When navigating the internet for media and entertainment, maintaining digital privacy is a priority. Here are a few tips:
The television series Sex Education has become a significant cultural touchpoint since its debut, praised for its nuanced portrayal of adolescence, identity, and interpersonal relationships. For those looking to watch the series, it is important to prioritize security and support the creators by using official channels. Overview of the Series