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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Content creators use sudden dhol beats to transition from a casual western look to a glamorous traditional Indian outfit.
The "Hindi Dhool Top"—commonly associated with the vibrant (drum) culture of India—is a style of attire designed for movement, celebration, and festive energy. These tops are often worn during weddings, Holi, and Punjabi-influenced festivities where the heavy beat of the dhol is central. Indian bride's wedding lengha. | Photo 226024 Maharani Weddings
One Diwali evening, the town held its annual kite battle. Elders bet on masala chai , kids bet on marbles. The sky filled with kites like a broken mirror of colors. Chhotu stood on his dhool top , feet sinking slightly into the warm earth, and released his Lal Patang — Red Kite.
No "Hindi Dhool Top" list is complete without the leading ladies of playback singing.
In the music world, "Dhool" is most famously associated with Adnan Dhool , the lead singer of the band
The term "Dhool Top" refers to the pinnacle of dhol-driven music, specifically in Hindi (Bollywood) and fusion styles. It encompasses songs that rely on a strong, percussive rhythm designed to make people dance.
A prime example is the highly-rated poetry book , available on platforms like Amazon India. This collection of ghazals, nazms, and shayari captures themes of love, longing, and pain. It showcases how an earthy, unfiltered approach to writing can touch the hearts of readers and climb to the top of digital book charts. Summary: The Ultimate "Dhool Top" Playlists & Collections
: Modern ethnic crop tops paired with dhoti pants or lehengas for dance festivals.
अमित: "कल का इंडिया-पाकिस्तान मैच देखा? क्या मैच था!" रोहन: "वो मैच नहीं, क़त्लेआम था! हमने तो उन्हें पूरी तरह धूल चटा दी ।" अमित: "बिल्कुल। हमारे गेंदबाज़ों ने तो उनके बल्लेबाज़ों को धूल में मिला दिया ।"
could refer to a type of clothing or could be interpreted in the context of rankings or preferences.
Upbeat tracks curated specifically for the groom or bride’s grand entrance and energetic sangeet performances. 3. Digital Trends: Viral Media and Content Creation
: A high-octane Punjabi-Hindi fusion track perfect for weddings.
"Coming from a small village with no resources, the famous actress is truly a flower of dust." एक छोटे से गाँव से आकर, यह प्रसिद्ध अभिनेत्री सच में एक धूल का फूल है।
The search query intersects three major cultural and linguistic elements: the Hindi language, the concept of "Dhool" (which translates to dust or earth in Hindi), and the concept of "Top" charts, musical hits, or high-end fashion trends.
Start with energetic, medium-paced songs and peak with the fastest, most rhythmic tracks.
Content creators use sudden dhol beats to transition from a casual western look to a glamorous traditional Indian outfit.
The "Hindi Dhool Top"—commonly associated with the vibrant (drum) culture of India—is a style of attire designed for movement, celebration, and festive energy. These tops are often worn during weddings, Holi, and Punjabi-influenced festivities where the heavy beat of the dhol is central. Indian bride's wedding lengha. | Photo 226024 Maharani Weddings hindi dhool top
One Diwali evening, the town held its annual kite battle. Elders bet on masala chai , kids bet on marbles. The sky filled with kites like a broken mirror of colors. Chhotu stood on his dhool top , feet sinking slightly into the warm earth, and released his Lal Patang — Red Kite.
No "Hindi Dhool Top" list is complete without the leading ladies of playback singing.
In the music world, "Dhool" is most famously associated with Adnan Dhool , the lead singer of the band The search query intersects three major cultural and
The term "Dhool Top" refers to the pinnacle of dhol-driven music, specifically in Hindi (Bollywood) and fusion styles. It encompasses songs that rely on a strong, percussive rhythm designed to make people dance.
A prime example is the highly-rated poetry book , available on platforms like Amazon India. This collection of ghazals, nazms, and shayari captures themes of love, longing, and pain. It showcases how an earthy, unfiltered approach to writing can touch the hearts of readers and climb to the top of digital book charts. Summary: The Ultimate "Dhool Top" Playlists & Collections
: Modern ethnic crop tops paired with dhoti pants or lehengas for dance festivals. The "Hindi Dhool Top"—commonly associated with the vibrant
अमित: "कल का इंडिया-पाकिस्तान मैच देखा? क्या मैच था!" रोहन: "वो मैच नहीं, क़त्लेआम था! हमने तो उन्हें पूरी तरह धूल चटा दी ।" अमित: "बिल्कुल। हमारे गेंदबाज़ों ने तो उनके बल्लेबाज़ों को धूल में मिला दिया ।"
could refer to a type of clothing or could be interpreted in the context of rankings or preferences.
Upbeat tracks curated specifically for the groom or bride’s grand entrance and energetic sangeet performances. 3. Digital Trends: Viral Media and Content Creation
: A high-octane Punjabi-Hindi fusion track perfect for weddings.
"Coming from a small village with no resources, the famous actress is truly a flower of dust." एक छोटे से गाँव से आकर, यह प्रसिद्ध अभिनेत्री सच में एक धूल का फूल है।