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/).
: Dual paddle triggers for instant software effect activation; toggle them up for "lock-on" or down for momentary "hold" effects. Performance Pads
Upon connecting, a detection window appears in VirtualDJ, allowing you to configure the audio routing for speakers and headphones. 2. Core Feature Performance Jog Wheels & Displays:
To unlock the full potential of your controller, pairing it with VirtualDJ creates a powerhouse setup for both bedroom DJs and working professionals.
: The unit is class-compliant, meaning Windows and macOS should recognize it automatically without manual driver installation. However, ensure your device is running the latest firmware (v1.10 or higher) to avoid mapping issues with the "Scratch" button.
The Numark Mixtrack Platinum FX is a 4-deck DJ controller designed to bridge the gap between entry-level gear and club-standard layouts. Key physical features include:
While the controller is optimized for Serato DJ Lite (and a license is included), many users prefer Virtual DJ for its innovative features and flexibility. Virtual DJ 2025's standout feature is the , which allows for on-the-fly remixing by recording and swapping isolated stems (like drums, vocals, or melody) of any track. This is a huge creative advantage. Furthermore, the virtualdj community praises its intuitive interface and advanced library management.
: These capacitive-touch platters feel weighted and professional. They feature high-resolution built-in color displays
While sturdy, the unit is entirely plastic, which may not hold up to intense, daily club use.
: Dual paddle triggers for instant software effect activation; toggle them up for "lock-on" or down for momentary "hold" effects. Performance Pads
Upon connecting, a detection window appears in VirtualDJ, allowing you to configure the audio routing for speakers and headphones. 2. Core Feature Performance Jog Wheels & Displays:
To unlock the full potential of your controller, pairing it with VirtualDJ creates a powerhouse setup for both bedroom DJs and working professionals.
: The unit is class-compliant, meaning Windows and macOS should recognize it automatically without manual driver installation. However, ensure your device is running the latest firmware (v1.10 or higher) to avoid mapping issues with the "Scratch" button.
The Numark Mixtrack Platinum FX is a 4-deck DJ controller designed to bridge the gap between entry-level gear and club-standard layouts. Key physical features include:
While the controller is optimized for Serato DJ Lite (and a license is included), many users prefer Virtual DJ for its innovative features and flexibility. Virtual DJ 2025's standout feature is the , which allows for on-the-fly remixing by recording and swapping isolated stems (like drums, vocals, or melody) of any track. This is a huge creative advantage. Furthermore, the virtualdj community praises its intuitive interface and advanced library management.
: These capacitive-touch platters feel weighted and professional. They feature high-resolution built-in color displays
While sturdy, the unit is entirely plastic, which may not hold up to intense, daily club use.