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/).
Rudra recovers his memory and returns to India for a final showdown with BBS on the banks of the Ganga. Season Ending
While terms like "WEB-DL" are frequently used in filesharing communities, downloading copyrighted content from unauthorized third-party websites exposes your device to malware, phishing risks, and legal vulnerabilities.
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: While some found it brilliant, many regular viewers gave it lower scores (around ) for being a let-down compared to Season 1 Key Highlights Rudra Shrivastava
as Ranjana Srivastava: Rudra's sharp-tongued wife.
The iconic dialogues and background music are best experienced with original, high-quality Hindi audio. Where to Watch and Download
Apharan Season 2: The Ultimate Guide to the Action-Thriller Sequel
These numbers refer to the video's resolution, which is essentially the number of pixels that make up the picture. More pixels mean a sharper, more detailed image.
Here’s a short, engaging post you can use:
The Indian web series scene has witnessed a significant surge in popularity over the past few years, with numerous shows captivating audiences across the country. One such show that has garnered a massive following is Apharan, a gripping web series that premiered with a bang and left viewers craving for more. The good news is that the wait is finally over, as Apharan Season 2 has arrived, and it's available for download in various resolutions, including WEB-DL Hindi 1080p, 720p, and 4K.
Production insights regarding the Share public link
Use headphones or a dedicated soundbar to fully appreciate the show's retro-themed background score and punchy dialogues. If you want to know more about the show, tell me:
WEB-DL files generally preserve advanced audio formats like Dolby Digital Plus 5.1, providing an immersive surround-sound experience for home theaters. Resolution Breakdown: 1080p, 720p, and 4K
Apharan Season 2 Web-DL: The Ultimate Guide to Arunoday Singh's Action-Thriller
: Often provides free, ad-supported viewing options. Airtel Xstream Play : Available for eligible subscribers. Series Overview
| Resolution | Aspect Ratio | Quality Level | | :--- | :--- | :--- | | | 16:9 | Very High: The standard for crisp, clear detail. The image contains 1080 vertical lines of pixels. | | 720p (HD Ready) | 16:9 | High: A good balance of quality and file size. It contains 720 vertical lines of pixels. | | 4K (Ultra HD) | 16:9 | Highest: Offers four times the resolution of 1080p. (Note: Availability of 4K may vary depending on source and platform). |
Rudra recovers his memory and returns to India for a final showdown with BBS on the banks of the Ganga. Season Ending
While terms like "WEB-DL" are frequently used in filesharing communities, downloading copyrighted content from unauthorized third-party websites exposes your device to malware, phishing risks, and legal vulnerabilities.
A WEB-DL (Web Download) is a file accurately monitored and ripped from a legal streaming platform (such as ALTBalaji or Voot Select). Unlike screen recordings, WEB-DL files undergo no re-encoding. This means you get the exact video and audio quality intended by the creators, free from on-screen watermarks or logos. 1080p vs. 720p Resolution WEB-DL 1080p WEB-DL 720p 1920 x 1080 pixels (Full HD) 1280 x 720 pixels (HD) Clarity Razor-sharp text, highly detailed textures Standard high-definition clarity File Size Larger (requires more storage and high speed) Compact (ideal for mobile devices) Best Used For Big screens, TVs, and desktop monitors Smartphones, tablets, and limited data Why Apharan Season 2 is a Must-Watch
: While some found it brilliant, many regular viewers gave it lower scores (around ) for being a let-down compared to Season 1 Key Highlights Rudra Shrivastava
as Ranjana Srivastava: Rudra's sharp-tongued wife. Apharan -Season 2- WEB-DL Hindi 1080p 720p 4...
The iconic dialogues and background music are best experienced with original, high-quality Hindi audio. Where to Watch and Download
Apharan Season 2: The Ultimate Guide to the Action-Thriller Sequel
These numbers refer to the video's resolution, which is essentially the number of pixels that make up the picture. More pixels mean a sharper, more detailed image.
Here’s a short, engaging post you can use: Rudra recovers his memory and returns to India
The Indian web series scene has witnessed a significant surge in popularity over the past few years, with numerous shows captivating audiences across the country. One such show that has garnered a massive following is Apharan, a gripping web series that premiered with a bang and left viewers craving for more. The good news is that the wait is finally over, as Apharan Season 2 has arrived, and it's available for download in various resolutions, including WEB-DL Hindi 1080p, 720p, and 4K.
Production insights regarding the Share public link
Use headphones or a dedicated soundbar to fully appreciate the show's retro-themed background score and punchy dialogues. If you want to know more about the show, tell me:
WEB-DL files generally preserve advanced audio formats like Dolby Digital Plus 5.1, providing an immersive surround-sound experience for home theaters. Resolution Breakdown: 1080p, 720p, and 4K 720p Resolution WEB-DL 1080p WEB-DL 720p 1920 x
Apharan Season 2 Web-DL: The Ultimate Guide to Arunoday Singh's Action-Thriller
: Often provides free, ad-supported viewing options. Airtel Xstream Play : Available for eligible subscribers. Series Overview
| Resolution | Aspect Ratio | Quality Level | | :--- | :--- | :--- | | | 16:9 | Very High: The standard for crisp, clear detail. The image contains 1080 vertical lines of pixels. | | 720p (HD Ready) | 16:9 | High: A good balance of quality and file size. It contains 720 vertical lines of pixels. | | 4K (Ultra HD) | 16:9 | Highest: Offers four times the resolution of 1080p. (Note: Availability of 4K may vary depending on source and platform). |