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/).
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Are you looking at a from that date?
While the slow burn remains popular, it is now less about delaying the physical relationship and more about building deep intellectual and emotional intimacy before the romance blooms [2]. 2. Redefining Romantic Tropes for the Modern Era
What is this for? (e.g., SEO blog, social media script, academic essay)
Mark your celestial calendars for . This is the anchor for the "24 08 28" keyword. On this specific Tuesday, the sky tells a dramatic story of transformation.
In the architecture of human connection, numbers often whisper what words cannot. The digits , 08 , and 28 form a fascinating love triangle of stability, emotion, and legacy. Whether read as birth dates, symbolic cycles, or character blueprints, these numbers tell a story of how different frequencies of love either harmonize or clash.
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This evolution allows writers to explore deeper themes, such as how mental health, financial stress, and career ambitions impact a partnership. By grounding characters in reality, the emotional stakes feel significantly higher for the audience. Inclusivity and Diverse Dynamics
: Showing couples actively resolving conflicts through dialogue.
August 28, 2024, was a day defined by a push-and-pull between deep spiritual longing and the heavy weight of earthly obligations. Whether viewed through the lens of astrology or the tropes of romantic literature, this date captures a specific type of "high-stakes" intimacy where partners must choose between the comfort of the status quo and the vulnerability of a "soulmate" connection. The Cosmic Conflict: Duty vs. Desire
The "Sexmex The Musical" episode is a perfect example of SexMex's innovative approach. Here's what it likely entails:
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Storylines now focus heavily on communication struggles, financial stress, and finding balance, showing that love isn't just a feeling, but a daily choice. 2. Redefining "Happily Ever After" (HEA)