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/).
For many readers, these stories serve as a form of emotional escapism. The rhythmic flow of the Tamil language, with its unique honorifics and terms of endearment, adds a layer of warmth to the prose that is hard to replicate in other languages. Phrases like "En Uyir Amma" or "Chella Magan" carry a weight of sentiment that anchors the reader to the characters' journey. Writers in this genre often use descriptive imagery—the scent of home-cooked food, the quiet of a shared evening, or the unspoken understanding in a glance—to build a world that feels both intimate and relatable.
The allure of a well-curated collection of these stories lies in the variety of themes they cover. Some narratives focus on the "Veettu Vishesham" style of family drama, where the son acts as the primary emotional anchor for a mother navigating the challenges of life. Others lean into a more protective "Kavalan" dynamic, where the son’s romanticized devotion manifests as an unwavering shield against the world’s hardships. The "romantic" element in these stories often refers to the idealized, poetic, and deeply affectionate way the characters interact, emphasizing a soul-connection that transcends ordinary household dynamics.
Amma magan tamil romantic fiction and stories collection The landscape of Tamil literature has always been rich with emotional depth, exploring complex human relationships and societal bonds. Within the vast realm of digital storytelling and contemporary fiction, one specific niche that has seen a significant surge in interest is the "Amma Magan" Tamil romantic fiction and stories collection. These stories, often characterized by their focus on the intricate, protective, and deeply emotional bond between a mother and her son, have found a dedicated readership across various online platforms and literary forums.
For many readers, these stories serve as a form of emotional escapism. The rhythmic flow of the Tamil language, with its unique honorifics and terms of endearment, adds a layer of warmth to the prose that is hard to replicate in other languages. Phrases like "En Uyir Amma" or "Chella Magan" carry a weight of sentiment that anchors the reader to the characters' journey. Writers in this genre often use descriptive imagery—the scent of home-cooked food, the quiet of a shared evening, or the unspoken understanding in a glance—to build a world that feels both intimate and relatable.
The allure of a well-curated collection of these stories lies in the variety of themes they cover. Some narratives focus on the "Veettu Vishesham" style of family drama, where the son acts as the primary emotional anchor for a mother navigating the challenges of life. Others lean into a more protective "Kavalan" dynamic, where the son’s romanticized devotion manifests as an unwavering shield against the world’s hardships. The "romantic" element in these stories often refers to the idealized, poetic, and deeply affectionate way the characters interact, emphasizing a soul-connection that transcends ordinary household dynamics.
Amma magan tamil romantic fiction and stories collection The landscape of Tamil literature has always been rich with emotional depth, exploring complex human relationships and societal bonds. Within the vast realm of digital storytelling and contemporary fiction, one specific niche that has seen a significant surge in interest is the "Amma Magan" Tamil romantic fiction and stories collection. These stories, often characterized by their focus on the intricate, protective, and deeply emotional bond between a mother and her son, have found a dedicated readership across various online platforms and literary forums.