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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The series is known for its intense intimate scenes, heightened by the magical and life-or-death stakes. The romantic moments between the lead characters are often described by viewers as a significant highlight, driving the "hot web series" label. 2. Digital Availability (JioHotstar)
: Also listed as a streaming provider in certain regions.
Mira must convince a "possessed" Aryan to break the bond himself, as a Daayan’s power can only be severed if the victim stops desiring the curse. Episodes (Mini-Series Format)
Aryan’s health begins to fade as his company’s stock soars.
If you’re searching for a gripping, bold Indian web series with supernatural or thriller elements (like the original Nazar TV show, but spicier), here are legitimate alternatives available today:
Nazar is primarily hosted on region-specific platforms. If you see a "Content not available in your country" error: Use a premium, reliable VPN service.
On the raw footage, the scene was quick—a fleeting moment where Ansh grabs Piya’s hand to save her from a falling debris. In the original broadcast, it lasted exactly two seconds before cutting to the villain’s reaction shot.
Under the guise of an interior designer, Ishika enters Aryan’s life. The series highlights the magnetic pull of her nazar , where every glance and touch slowly weakens Aryan's willpower while boosting his professional success through dark magic.
Shortly after the release of “Chhupi Nazar” Part 1, various illegal piracy websites—such as —leaked the full episode download links. When users search for “Nazar hot web series fixed,” they are often looking for a fixed or working download link that hasn’t been taken down or broken by copyright enforcement.
🛡️ Beware of Scam Sites: Finding the Real Nazar Web Series
"Nazar" is part of the growing library of adult and thriller-themed content on regional OTT (Over-The-Top) platforms. Typically, these series aim to cater to a demographic looking for intense drama, suspense, and intimate, high-stakes scenarios. The series often revolves around themes of desire, betrayal, or intense relationships, leading to scenes that are considered "hot" or explicit by viewers.
She pressed it to the child's forehead in a movement as old as lullabies. People murmured, some with reverence, some with suspicion. The child’s breaths steadied. The mother’s hands found Rukmini’s like a lifeline and refused to let go.