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/).
If you are looking to add a "night high series" to your digital library, start with . Kobo often bundles the "Reze Arc" movies and the manga volumes, offering them in a high-quality digital format perfect for reading on the Kobo Clara BW or Libra Colour e-readers. The series is the benchmark for modern night-high aesthetics.
The keyword "night high series denji kobo top" is an unsolved riddle. Here is a checklist to help you solve it:
In true Denji Kobo fashion, the branding is whispered, not shouted. You’ll find tonal embroidery and custom hardware that speak to those "in the know," making it a staple for anyone who prefers quiet luxury over loud logos. How to Style the Night High Series Top
In the Japanese indie scene, a kobo (工房) functions as a small workshop or studio. Denji Kobo operated as a classic dōjin circle, meaning their titles were produced by a handful of passionate developers, writers, and pixel artists.
So, if you're searching for that "night high" experience, you've come to the right place. You're looking for a series that demands your attention and offers pure, unfiltered catharsis. Let's get into why Chainsaw Man is exactly that series.
Certain Denji Kobo models incorporate specialized phosphorescent (glow-in-the-dark) materials, ensuring that even if the water is pitch black, the lure remains a target. 3. Precision Engineering and Balance
What makes Denji so refreshingly unique is his moral ambiguity and his unashamed, primal desires. He isn't driven by justice or a code of ethics. His motivations often boil down to the basest of human instincts: food, shelter, and physical affection. He gets recruited into a government Devil Hunting agency not out of patriotism, but because he is bribed with the promise of a comfortable life and the affection of the beautiful, enigmatic Makima. This subversion of the classic "chosen one" narrative is part of the series' genius. Denji is a genuinely good person at heart, but he's also selfish, confused, and constantly manipulated, making him one of the most relatable and unpredictable protagonists in modern manga.
The Night High Series, also known as the "Denji Kobo" series, has gained significant attention in recent years, particularly among anime enthusiasts. This Japanese light novel series, written by Kōta Kōno and illustrated by Tsukumizu, has been adapted into several manga and anime formats. In this post, we'll dive into the world of Night High Series: Denji Kobo Top, exploring its intricate storylines, memorable characters, and themes.
: Enthusiasts can find archived builds of the series, such as the digital catalog hosted on the Internet Archive Night High Section , which preserves the files for historical research and emulation.
Of course, we must return to the source. "Denji Kobo Top" is literally synonymous with Chainsaw Man on the platform. On Rakuten Kobo, Chainsaw Man consistently ranks in the , alongside giants like Kingdom and Attack on Titan .
Night High! : Denji kobo : Free Download, Borrow, and Streaming : Internet Archive. Internet Archive
When fans discuss the "top" aspects of the Night High Series, they are usually referring to the standout releases, iconic character archetypes, and defining artistic choices that set this series apart from its contemporary peers. Top Visual Motifs
The Night High Series represents a focused lineup of specialized lures and tackle designed for nighttime fishing scenarios. These tools are engineered to be highly effective when light is low and predatory fish are relying on senses other than sight, such as vibration and sound.
For the voracious reader looking to recreate that "night high" feeling every single day, is a game-changer.
Keep sharp objects like keycap pullers at a safe angle when swapping out switches to avoid scraping the custom interior walls of the top housing.
This comprehensive guide explores the structural elements, origins, artistic themes, and procurement avenues for the Night High Series by Denji Kobo.
If you are looking to add a "night high series" to your digital library, start with . Kobo often bundles the "Reze Arc" movies and the manga volumes, offering them in a high-quality digital format perfect for reading on the Kobo Clara BW or Libra Colour e-readers. The series is the benchmark for modern night-high aesthetics.
The keyword "night high series denji kobo top" is an unsolved riddle. Here is a checklist to help you solve it:
In true Denji Kobo fashion, the branding is whispered, not shouted. You’ll find tonal embroidery and custom hardware that speak to those "in the know," making it a staple for anyone who prefers quiet luxury over loud logos. How to Style the Night High Series Top
In the Japanese indie scene, a kobo (工房) functions as a small workshop or studio. Denji Kobo operated as a classic dōjin circle, meaning their titles were produced by a handful of passionate developers, writers, and pixel artists.
So, if you're searching for that "night high" experience, you've come to the right place. You're looking for a series that demands your attention and offers pure, unfiltered catharsis. Let's get into why Chainsaw Man is exactly that series. night high series denji kobo top
Certain Denji Kobo models incorporate specialized phosphorescent (glow-in-the-dark) materials, ensuring that even if the water is pitch black, the lure remains a target. 3. Precision Engineering and Balance
What makes Denji so refreshingly unique is his moral ambiguity and his unashamed, primal desires. He isn't driven by justice or a code of ethics. His motivations often boil down to the basest of human instincts: food, shelter, and physical affection. He gets recruited into a government Devil Hunting agency not out of patriotism, but because he is bribed with the promise of a comfortable life and the affection of the beautiful, enigmatic Makima. This subversion of the classic "chosen one" narrative is part of the series' genius. Denji is a genuinely good person at heart, but he's also selfish, confused, and constantly manipulated, making him one of the most relatable and unpredictable protagonists in modern manga.
The Night High Series, also known as the "Denji Kobo" series, has gained significant attention in recent years, particularly among anime enthusiasts. This Japanese light novel series, written by Kōta Kōno and illustrated by Tsukumizu, has been adapted into several manga and anime formats. In this post, we'll dive into the world of Night High Series: Denji Kobo Top, exploring its intricate storylines, memorable characters, and themes.
: Enthusiasts can find archived builds of the series, such as the digital catalog hosted on the Internet Archive Night High Section , which preserves the files for historical research and emulation. If you are looking to add a "night
Of course, we must return to the source. "Denji Kobo Top" is literally synonymous with Chainsaw Man on the platform. On Rakuten Kobo, Chainsaw Man consistently ranks in the , alongside giants like Kingdom and Attack on Titan .
Night High! : Denji kobo : Free Download, Borrow, and Streaming : Internet Archive. Internet Archive
When fans discuss the "top" aspects of the Night High Series, they are usually referring to the standout releases, iconic character archetypes, and defining artistic choices that set this series apart from its contemporary peers. Top Visual Motifs
The Night High Series represents a focused lineup of specialized lures and tackle designed for nighttime fishing scenarios. These tools are engineered to be highly effective when light is low and predatory fish are relying on senses other than sight, such as vibration and sound. The keyword "night high series denji kobo top"
For the voracious reader looking to recreate that "night high" feeling every single day, is a game-changer.
Keep sharp objects like keycap pullers at a safe angle when swapping out switches to avoid scraping the custom interior walls of the top housing.
This comprehensive guide explores the structural elements, origins, artistic themes, and procurement avenues for the Night High Series by Denji Kobo.