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/).
Granger’s approach is unique because it forces students to think like engineers through three distinct stages:
: Analyze the physical aspects to understand how the system works in the real world.
The book "Fluid Mechanics" by Granger is a comprehensive textbook that covers the fundamental principles of fluid mechanics, including:
Unfortunately, I couldn't find a publicly available PDF link for "Fluid Mechanics" by Granger due to copyright restrictions. However, I can suggest some alternatives:
Examination of laminar and turbulent flow behaviors. 💡 Key Insight
If you are looking for more accessible digital guides on the same principles: A Guide to Fluid Mechanics (Hongwei Wang)
While they may not have Granger’s specific book, MIT OCW offers complete, free downloadable PDFs of fluid mechanics lecture notes, textbooks, and problem sets.
Robert A. Granger's Fluid Mechanics is a cornerstone text for anyone serious about understanding fluid dynamics. Its unique pedagogical method, clear prose, and extensive illustrations have earned it praise from both students and professionals. While legitimate PDF copies are available for purchase or through library portals, the vast majority of "free PDF" links online are unreliable, spam-filled, or illegal.
, academic institutions and libraries often provide guides or partial PDFs for study purposes: Fluid Mechanics Robert A. Granger Guide (UML) - A summary guide on mastering his core principles. NJIT Syllabus & Reading List
" can be difficult as the book is a copyrighted publication by Courier Corporation.
A non-profit digital library offering millions of free books. You can often legally borrow a digital copy of Fluid Mechanics by Robert A. Granger for 1-hour or 14-day increments.
The fundamental theories detailed in the text directly translate to modern engineering disciplines: Fluid Mechanics - MAE Princeton
Dynamics introduces the relationship between fluid motion and the forces acting upon it. This is where the heavy mathematics of engineering come into play, governed by three fundamental conservation laws:
Granger’s approach is unique because it forces students to think like engineers through three distinct stages:
: Analyze the physical aspects to understand how the system works in the real world.
The book "Fluid Mechanics" by Granger is a comprehensive textbook that covers the fundamental principles of fluid mechanics, including:
Unfortunately, I couldn't find a publicly available PDF link for "Fluid Mechanics" by Granger due to copyright restrictions. However, I can suggest some alternatives:
Examination of laminar and turbulent flow behaviors. 💡 Key Insight
If you are looking for more accessible digital guides on the same principles: A Guide to Fluid Mechanics (Hongwei Wang)
While they may not have Granger’s specific book, MIT OCW offers complete, free downloadable PDFs of fluid mechanics lecture notes, textbooks, and problem sets.
Robert A. Granger's Fluid Mechanics is a cornerstone text for anyone serious about understanding fluid dynamics. Its unique pedagogical method, clear prose, and extensive illustrations have earned it praise from both students and professionals. While legitimate PDF copies are available for purchase or through library portals, the vast majority of "free PDF" links online are unreliable, spam-filled, or illegal.
, academic institutions and libraries often provide guides or partial PDFs for study purposes: Fluid Mechanics Robert A. Granger Guide (UML) - A summary guide on mastering his core principles. NJIT Syllabus & Reading List
" can be difficult as the book is a copyrighted publication by Courier Corporation.
A non-profit digital library offering millions of free books. You can often legally borrow a digital copy of Fluid Mechanics by Robert A. Granger for 1-hour or 14-day increments.
The fundamental theories detailed in the text directly translate to modern engineering disciplines: Fluid Mechanics - MAE Princeton
Dynamics introduces the relationship between fluid motion and the forces acting upon it. This is where the heavy mathematics of engineering come into play, governed by three fundamental conservation laws: