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/).
Every year has its "it" people, and 2021 was no different. The entertainment scene was invigorated by fresh faces whose photos went viral daily:
Instead of fine dining in Paris, 2021 photos showed actors like eating at legendary warteg (street stalls) in Tangerang or Bandung. The photo of Raffi holding a cracked phone case while eating tahu gejrot became a meme and a lifestyle statement: "I am rich, but I am also merakyat (down to earth)."
The year 2021 was a transformative period for the Indonesian entertainment industry. Despite global challenges, the "lifestyle and entertainment" scene thrived through digital innovation and the rise of versatile stars who dominated both the silver screen and social media.
The young actress faced significant public scrutiny when a video involving intimate behavior with her then-boyfriend surfaced online, marking her second major social media controversy within a short period.
The lifestyle of Indonesian actors has undergone significant changes in recent years, with many embracing a more modern and healthy approach to living. Poto Aktor Indonesia 2021 showcased the daily routines of various actors, revealing their fitness regimens, dietary habits, and personal interests.
In this deep dive, we explore how these snapshots became more than just images; they became a visual diary of resilience, fashion evolution, and the shifting landscape of .
| Actor | Notable 2021 Projects | Lifestyle Highlights | |-------|----------------------|----------------------| | | Layla Majnun , Waktu Maghrib | Minimalist fashion, reading, film festival appearances | | Chicco Jerikho | Seperti Dendam, Rindu Harus Dibayar Tuntas | Outdoor lifestyle, hiking, sustainable fashion | | Joe Taslim | Mortal Kombat (Hollywood), Serigala Terakhir | Martial arts training, family time, streetwear | | Iqbaal Ramadhan | Ali & Ratu Ratu Queens | Music (band Svmmerdose), skateboarding, casual youth style | | Arbani Yasiz | Modeling, Satu Hari yang Panjang | Gym workouts, luxury travel, sneaker collection | | Angga Yunanda | Mariposa , Buka Zaman Itu | Romantic BTS shots, casual fits, friendship with Shenina |
Every year has its "it" people, and 2021 was no different. The entertainment scene was invigorated by fresh faces whose photos went viral daily:
Instead of fine dining in Paris, 2021 photos showed actors like eating at legendary warteg (street stalls) in Tangerang or Bandung. The photo of Raffi holding a cracked phone case while eating tahu gejrot became a meme and a lifestyle statement: "I am rich, but I am also merakyat (down to earth)."
The year 2021 was a transformative period for the Indonesian entertainment industry. Despite global challenges, the "lifestyle and entertainment" scene thrived through digital innovation and the rise of versatile stars who dominated both the silver screen and social media.
The young actress faced significant public scrutiny when a video involving intimate behavior with her then-boyfriend surfaced online, marking her second major social media controversy within a short period.
The lifestyle of Indonesian actors has undergone significant changes in recent years, with many embracing a more modern and healthy approach to living. Poto Aktor Indonesia 2021 showcased the daily routines of various actors, revealing their fitness regimens, dietary habits, and personal interests.
In this deep dive, we explore how these snapshots became more than just images; they became a visual diary of resilience, fashion evolution, and the shifting landscape of .
| Actor | Notable 2021 Projects | Lifestyle Highlights | |-------|----------------------|----------------------| | | Layla Majnun , Waktu Maghrib | Minimalist fashion, reading, film festival appearances | | Chicco Jerikho | Seperti Dendam, Rindu Harus Dibayar Tuntas | Outdoor lifestyle, hiking, sustainable fashion | | Joe Taslim | Mortal Kombat (Hollywood), Serigala Terakhir | Martial arts training, family time, streetwear | | Iqbaal Ramadhan | Ali & Ratu Ratu Queens | Music (band Svmmerdose), skateboarding, casual youth style | | Arbani Yasiz | Modeling, Satu Hari yang Panjang | Gym workouts, luxury travel, sneaker collection | | Angga Yunanda | Mariposa , Buka Zaman Itu | Romantic BTS shots, casual fits, friendship with Shenina |