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/).
Indian culture is not a static museum piece; it is a living, breathing entity. It is a land where cows roam freely near high-tech IT hubs and where the latest pop music plays alongside the ancient echoes of a Sitar. To embrace the Indian lifestyle is to embrace contradictions, vibrant colors, and an unwavering sense of hope.
The most defining characteristic of Indian culture is its pluralism. India is home to nearly every major religion in the world, hundreds of languages, and thousands of dialects. Yet, a shared "Indianness" binds the population. This lifestyle is built on the Vedic philosophy of Vasudhaiva Kutumbakam —the world is one family. 2. The Social Fabric: Family and Community In India, life is rarely lived in isolation.
The Saree, often called the world's oldest unstitched garment, remains a symbol of grace. Similarly, the Salwar Kameez and Kurta-Pajama offer comfort across the subcontinent.
Festivals like Diwali, Eid, Holi, and Christmas are celebrated across communal lines. The "neighborhood culture" is strong; it’s common for neighbors to share meals and participate in each other’s life milestones. 3. Culinary Traditions: More Than Just Spice Indian food is a sensory map of the country’s geography.
Today’s Indian culture is as much about Silicon Valley as it is about the Ganges.
With one of the world's largest smartphone-user bases, daily life in India—from ordering groceries to finding a life partner—happens on apps.
Indian lifestyle content is incomplete without mentioning its sartorial elegance.
Here is an in-depth look at the pillars of Indian culture and how they shape daily life today. 1. The Core Philosophy: Unity in Diversity
India is the birthplace of Yoga and Meditation, practices that have now become global wellness phenomena. For many Indians, spirituality is integrated into the daily routine:
Indian culture is a kaleidoscope of traditions, flavors, and values that have evolved over five millennia. To understand the lifestyle that stems from this heritage, one must look past the stereotypes and explore the intricate balance between ancient roots and a rapidly modernizing society.
Indian culture is not a static museum piece; it is a living, breathing entity. It is a land where cows roam freely near high-tech IT hubs and where the latest pop music plays alongside the ancient echoes of a Sitar. To embrace the Indian lifestyle is to embrace contradictions, vibrant colors, and an unwavering sense of hope.
The most defining characteristic of Indian culture is its pluralism. India is home to nearly every major religion in the world, hundreds of languages, and thousands of dialects. Yet, a shared "Indianness" binds the population. This lifestyle is built on the Vedic philosophy of Vasudhaiva Kutumbakam —the world is one family. 2. The Social Fabric: Family and Community In India, life is rarely lived in isolation.
The Saree, often called the world's oldest unstitched garment, remains a symbol of grace. Similarly, the Salwar Kameez and Kurta-Pajama offer comfort across the subcontinent.
Festivals like Diwali, Eid, Holi, and Christmas are celebrated across communal lines. The "neighborhood culture" is strong; it’s common for neighbors to share meals and participate in each other’s life milestones. 3. Culinary Traditions: More Than Just Spice Indian food is a sensory map of the country’s geography.
Today’s Indian culture is as much about Silicon Valley as it is about the Ganges.
With one of the world's largest smartphone-user bases, daily life in India—from ordering groceries to finding a life partner—happens on apps.
Indian lifestyle content is incomplete without mentioning its sartorial elegance.
Here is an in-depth look at the pillars of Indian culture and how they shape daily life today. 1. The Core Philosophy: Unity in Diversity
India is the birthplace of Yoga and Meditation, practices that have now become global wellness phenomena. For many Indians, spirituality is integrated into the daily routine:
Indian culture is a kaleidoscope of traditions, flavors, and values that have evolved over five millennia. To understand the lifestyle that stems from this heritage, one must look past the stereotypes and explore the intricate balance between ancient roots and a rapidly modernizing society.