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/).
Here is everything you need to know about watching , including its plot, cast, standout themes, and where you can stream it today. The Captivating Plot of The K2
No Indian drama is complete without a memorable villain, and Choi Yoo-jin (played brilliantly by Song Yoon-ah) is the gift that keeps on giving. The Hindi dub for Yoo-jin is particularly outstanding. Her manipulative whispers and sharp insults land with a piercing impact in Hindi. The voice actress captures the perfect blend of elegance and malice, reminding viewers of the iconic "Vamps" of Indian daily soaps, but with a sophisticated, high-fashion twist.
Upon returning to South Korea, Je-ha accidentally witnesses an extramarital affair involving Jang Se-joon (Jo Sung-ha), a prominent presidential candidate. To silence him, Choi Yoo-jin (Song Yoon-ah), Se-joon’s ambitious and ruthless wife, targets Je-ha.
Are you a fan of Korean dramas and looking for a thrilling adventure? Look no further than "The K2" drama in Hindi dubbed. This action-packed drama has gained immense popularity worldwide, and its Hindi dubbed version has made it accessible to a wider audience.
The dialogue is adapted to make the political intrigue and social hierarchies, which are central to the story, easily understandable to a Hindi-speaking audience. Key Characters and Performances
The world of "The K2" is dark, thrilling, and action-packed. Its availability in Hindi dubbed has only widened its fan base, proving that a well-made action drama has universal appeal. If you can find it, do not miss the chance to watch Kim Je-ha take on the world, one spectacular fight scene at a time.
Watching makes the action-packed scenes even more engaging. The dubbing allows viewers to focus entirely on the brilliant cinematography, intricate fight choreography, and emotional acting, rather than shifting their gaze between the action and the subtitles. Why the Hindi Dubbing Works:
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Even if you don’t find the perfect Hindi dub, The K2 deserves your time. Here is why:
Here is everything you need to know about watching , including its plot, cast, standout themes, and where you can stream it today. The Captivating Plot of The K2
No Indian drama is complete without a memorable villain, and Choi Yoo-jin (played brilliantly by Song Yoon-ah) is the gift that keeps on giving. The Hindi dub for Yoo-jin is particularly outstanding. Her manipulative whispers and sharp insults land with a piercing impact in Hindi. The voice actress captures the perfect blend of elegance and malice, reminding viewers of the iconic "Vamps" of Indian daily soaps, but with a sophisticated, high-fashion twist.
Upon returning to South Korea, Je-ha accidentally witnesses an extramarital affair involving Jang Se-joon (Jo Sung-ha), a prominent presidential candidate. To silence him, Choi Yoo-jin (Song Yoon-ah), Se-joon’s ambitious and ruthless wife, targets Je-ha.
Are you a fan of Korean dramas and looking for a thrilling adventure? Look no further than "The K2" drama in Hindi dubbed. This action-packed drama has gained immense popularity worldwide, and its Hindi dubbed version has made it accessible to a wider audience.
The dialogue is adapted to make the political intrigue and social hierarchies, which are central to the story, easily understandable to a Hindi-speaking audience. Key Characters and Performances
The world of "The K2" is dark, thrilling, and action-packed. Its availability in Hindi dubbed has only widened its fan base, proving that a well-made action drama has universal appeal. If you can find it, do not miss the chance to watch Kim Je-ha take on the world, one spectacular fight scene at a time.
Watching makes the action-packed scenes even more engaging. The dubbing allows viewers to focus entirely on the brilliant cinematography, intricate fight choreography, and emotional acting, rather than shifting their gaze between the action and the subtitles. Why the Hindi Dubbing Works:
Watch The K2 Web Series Online for free in HD | Amazon MX Player. Sorry, this content is currently not available. In the meantime, Amazon MX Player Watch The K2 | Netflix
Even if you don’t find the perfect Hindi dub, The K2 deserves your time. Here is why: