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
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The keyword you've provided seems to be a search query or a tag that might be used on various platforms or search engines. It's looking for images of a person named Luana, specifically from the ages of 11 to 13 years old. The query mentions "743 pics," suggesting a considerable number of images, and "18500m free," which could imply that the content is freely available or perhaps there's a specific file size or type (like a 18.5MB file) associated with it.
As technology continues to evolve, so too will the challenges and solutions related to digital privacy. It's crucial for individuals, communities, and governments to work together to create a safer online environment. This includes educating individuals about the importance of privacy, the potential risks of sharing personal content online, and the steps they can take to protect themselves.
I can create a comprehensive article based on your request. However, I want to emphasize the importance of respecting privacy, especially when it comes to minors. The keyword you've provided suggests a search for images of a specific individual, Luana, from 11 to 13 years old. It's crucial to approach this topic with sensitivity and legality in mind.
In today's digital world, the availability of information and images online has become unprecedented. With just a few clicks, one can access a vast array of content, including images, videos, and personal information about individuals. However, this ease of access raises significant concerns about privacy, especially concerning minors.
The keyword you provided highlights the complexities of digital privacy and the need for vigilance and responsibility online. While it's easy to get caught up in the vastness of digital content, it's essential to remember the individuals behind the images and information. By promoting a culture of respect, consent, and safety, we can help ensure a healthier digital environment for everyone.
If you're looking to learn more about online safety, privacy practices, or how to protect your digital footprint, there are numerous resources available online. Libraries, educational institutions, and cybersecurity experts often provide guidance and tools to help navigate these issues.