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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However, HiWebxSeries.com is a licensed, major OTT service like Amazon Prime, ZEE5, or MX Player. Instead, it falls into a gray area of the internet – platforms that curate and sometimes re-upload content without full copyright licenses.
In conclusion, verification is a critical process that ensures the security, trust, and accountability of online platforms. While there are challenges and limitations to verification, it is essential for platforms to implement robust verification processes to protect their users and maintain the integrity of their platforms.
The verification process typically involves providing identification documents, such as a passport or driver's license, and confirming the user's identity through various means, such as email or phone verification. Some platforms may also use advanced technologies, such as facial recognition or biometric authentication, to verify a user's identity.
Bhookh (2024 mini-series), for which she received critical acclaim.
Context and likely components
: She is recognized for her performances in series such as "Palang Tod" (2020), "61-62 Rocket" (2022), and "Bhookh" (2024). Verified Status
However, HiWebxSeries.com is a licensed, major OTT service like Amazon Prime, ZEE5, or MX Player. Instead, it falls into a gray area of the internet – platforms that curate and sometimes re-upload content without full copyright licenses.
In conclusion, verification is a critical process that ensures the security, trust, and accountability of online platforms. While there are challenges and limitations to verification, it is essential for platforms to implement robust verification processes to protect their users and maintain the integrity of their platforms.
The verification process typically involves providing identification documents, such as a passport or driver's license, and confirming the user's identity through various means, such as email or phone verification. Some platforms may also use advanced technologies, such as facial recognition or biometric authentication, to verify a user's identity.
Bhookh (2024 mini-series), for which she received critical acclaim.
Context and likely components