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/).
Several non-profit digital libraries allow you to "borrow" a digital copy legally:
Misleadingly labeled to attract traffic. Important Note on Digital Safety
The Search for " Juventud en Éxtasis 2 " PDF on Google Drive: What Readers Should Know
The dramatic, easy-to-read style made it accessible to teenagers and young adults, providing a moral compass in a fast-changing world. Searching for "Juventud en Éxtasis 2"
First published in the late 1990s, Juventud en Éxtasis 2 continues the educational and narrative mission of its predecessor. Carlos Cuauhtémoc Sánchez utilizes a blend of fiction and essay-like commentary to address the real-world dilemmas faced by young adults navigating love, temptation, and societal pressures. Core Themes of the Book
If you want to read Juventud en Éxtasis 2 safely while supporting the author, consider the following legitimate avenues:
It is presented as a "complementary didactic material" containing research, guides, and tools for young people to manage their sexual conduct and moral values.
Cómo decir "no" a las presiones de pareja o amigos sin sentirse culpable. Enlace al Libro (Resumen y Materiales)
The content of the book is valuable, aiming to help adolescents navigate their emotions and sexuality with dignity and responsibility. However, accessing it should be done responsibly. If you are interested in reading this book, the best recommendation is to purchase it from an official digital bookstore or a physical retailer. The wisdom contained in its pages is worth paying for.
Si encuentras útil este resumen, te recomiendo buscar la versión física o el E-book oficial en librerías digitales.
: A full PDF version is available for direct reading or download.
Exploring the Cultural Significance of "Juventud en Éxtasis 2"
Several non-profit digital libraries allow you to "borrow" a digital copy legally:
Misleadingly labeled to attract traffic. Important Note on Digital Safety
The Search for " Juventud en Éxtasis 2 " PDF on Google Drive: What Readers Should Know
The dramatic, easy-to-read style made it accessible to teenagers and young adults, providing a moral compass in a fast-changing world. Searching for "Juventud en Éxtasis 2"
First published in the late 1990s, Juventud en Éxtasis 2 continues the educational and narrative mission of its predecessor. Carlos Cuauhtémoc Sánchez utilizes a blend of fiction and essay-like commentary to address the real-world dilemmas faced by young adults navigating love, temptation, and societal pressures. Core Themes of the Book
If you want to read Juventud en Éxtasis 2 safely while supporting the author, consider the following legitimate avenues:
It is presented as a "complementary didactic material" containing research, guides, and tools for young people to manage their sexual conduct and moral values.
Cómo decir "no" a las presiones de pareja o amigos sin sentirse culpable. Enlace al Libro (Resumen y Materiales)
The content of the book is valuable, aiming to help adolescents navigate their emotions and sexuality with dignity and responsibility. However, accessing it should be done responsibly. If you are interested in reading this book, the best recommendation is to purchase it from an official digital bookstore or a physical retailer. The wisdom contained in its pages is worth paying for.
Si encuentras útil este resumen, te recomiendo buscar la versión física o el E-book oficial en librerías digitales.
: A full PDF version is available for direct reading or download.
Exploring the Cultural Significance of "Juventud en Éxtasis 2"