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/).
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"The Vibrant Tapestry of Indian Family Life: Stories from the Heart of India"
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Indian families typically start their day early, with a morning prayer or meditation session. The aroma of freshly cooked breakfast wafts through the air, often consisting of traditional dishes like idlis, dosas, or parathas. After breakfast, family members go about their daily routines, with children heading off to school and adults attending to their work or household chores. In many Indian households, the mother plays a pivotal role in managing the family and ensuring everyone's well-being. As Priya, a working mother from Delhi, says, "My day starts with a quick prayer and a cup of tea. I then help my children get ready for school and make sure they have a healthy breakfast before heading out the door."
Indian cuisine is famous for its diversity and richness, with a wide range of flavors and spices used in different regions. Family meals are an essential part of Indian life, often featuring traditional dishes like curries, biryani, and tandoori chicken. The Sunday lunch, in particular, is a special occasion, with families gathering together to share a hearty meal. As Nalini, a homemaker from Chennai, says, "Sunday lunch is a tradition in our family. We make a big spread of traditional dishes, and everyone gathers around the table to share stories and laughter."
In India, the joint family system is still prevalent, especially in rural areas. This system, where multiple generations live together under one roof, fosters a sense of unity, respect, and interdependence. Children learn valuable life lessons from their grandparents, while the elderly benefit from the care and companionship of their younger family members. For example, Rohan, a young boy from Mumbai, shares his experience of living with his grandparents: "I love listening to my grandfather's stories about our family's history and traditions. He teaches me about the importance of respect, hard work, and community service."
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"The Vibrant Tapestry of Indian Family Life: Stories from the Heart of India"
We hope you enjoyed this blog post! Let us know if you have any feedback or suggestions for future posts.
Indian families typically start their day early, with a morning prayer or meditation session. The aroma of freshly cooked breakfast wafts through the air, often consisting of traditional dishes like idlis, dosas, or parathas. After breakfast, family members go about their daily routines, with children heading off to school and adults attending to their work or household chores. In many Indian households, the mother plays a pivotal role in managing the family and ensuring everyone's well-being. As Priya, a working mother from Delhi, says, "My day starts with a quick prayer and a cup of tea. I then help my children get ready for school and make sure they have a healthy breakfast before heading out the door."
Indian cuisine is famous for its diversity and richness, with a wide range of flavors and spices used in different regions. Family meals are an essential part of Indian life, often featuring traditional dishes like curries, biryani, and tandoori chicken. The Sunday lunch, in particular, is a special occasion, with families gathering together to share a hearty meal. As Nalini, a homemaker from Chennai, says, "Sunday lunch is a tradition in our family. We make a big spread of traditional dishes, and everyone gathers around the table to share stories and laughter."
In India, the joint family system is still prevalent, especially in rural areas. This system, where multiple generations live together under one roof, fosters a sense of unity, respect, and interdependence. Children learn valuable life lessons from their grandparents, while the elderly benefit from the care and companionship of their younger family members. For example, Rohan, a young boy from Mumbai, shares his experience of living with his grandparents: "I love listening to my grandfather's stories about our family's history and traditions. He teaches me about the importance of respect, hard work, and community service."