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/).
Tariq’s phone buzzes. (Gianni Paolo): “Dude. They found Jabari.” Tariq doesn’t reply. He’s already late for his first law school study group.
: To move product on the Stansfield campus, Tariq partners with Simon Stern’s husband, Bash, to create "CourseCorrect" —a tutoring app that doubles as a front for drug deliveries.
Power Book II: Ghost EPISODE: Season 1, Episode 4 QUALITY: WEBRip 🎬
Focus: Technical details and file info.
[Link 1] [Link 2] [Magnet Link]
. Partnering with Sebastian Kamal-Stern, he creates a platform that delivers more than just study guides—it serves as the ultimate front for moving product directly to dorm rooms. By insulating himself and the Tejadas, Tariq successfully takes over the campus drug market, effectively pushing out his main rival, Scott, through a clever set-up at a party. The Tejada Family Turmoil Monet Tejada continues to rule with an iron fist, but the cracks are starting to show: The GTG Cutoff: After Lil Guap disrespects Monet, Cane delivers a brutal beating to the GTG crew, officially ending their partnership. Tariq the New Partner: Much to Cane’s vocal disapproval, Monet announces that Tariq will be their new distributor on campus. Secret Lives: Monet discovers Dru’s secret artistic side when she finds a sketch in his bag, though she chooses to lie and tell him she believes he is "capable" of running the business despite his father's doubts. Love Triangles and Double Agents Tariq’s personal life is getting as messy as his professional one. He shares a kiss with Lauren at a high-tension dinner with her snobby parents and professors, only to be interrupted by her "cheeseball" boyfriend, Malcolm. Meanwhile, he also shares a moment and a kiss with Diana Tejada when she delivers product to him on campus. Outside the campus bubble, Cooper Saxe is still on the hunt. He enlists his niece, Riley, to go undercover and spy on Tariq at Stansfield. She successfully snaps a photo of Tariq that could spell trouble down the line. Tasha’s Warning From behind bars, Tasha is growing increasingly paranoid. Sensing the danger her son is in, she instructs Tariq to find a gun and protect himself, even pointing him toward one of Ghost and Tommy’s old stash spots. As Tariq balances his Ivy League education with his rising status as a "Prince" of the drug trade, it's clear he's more like his father than he'd ever care to admit. Are you ready to see how
Tariq doesn’t flinch. But when he gets back to his dorm, (Method Man)—his slick, morally flexible attorney—is waiting. Davis drops a bombshell: the feds are building a Rico case using Jabari’s missing laptop. And Jabari had footage of Tariq visiting him the night he died.
Tariq’s phone buzzes. (Gianni Paolo): “Dude. They found Jabari.” Tariq doesn’t reply. He’s already late for his first law school study group.
: To move product on the Stansfield campus, Tariq partners with Simon Stern’s husband, Bash, to create "CourseCorrect" —a tutoring app that doubles as a front for drug deliveries. power book ii: ghost s01e04 webrip
Power Book II: Ghost EPISODE: Season 1, Episode 4 QUALITY: WEBRip 🎬 Tariq’s phone buzzes
Focus: Technical details and file info.
[Link 1] [Link 2] [Magnet Link]
. Partnering with Sebastian Kamal-Stern, he creates a platform that delivers more than just study guides—it serves as the ultimate front for moving product directly to dorm rooms. By insulating himself and the Tejadas, Tariq successfully takes over the campus drug market, effectively pushing out his main rival, Scott, through a clever set-up at a party. The Tejada Family Turmoil Monet Tejada continues to rule with an iron fist, but the cracks are starting to show: The GTG Cutoff: After Lil Guap disrespects Monet, Cane delivers a brutal beating to the GTG crew, officially ending their partnership. Tariq the New Partner: Much to Cane’s vocal disapproval, Monet announces that Tariq will be their new distributor on campus. Secret Lives: Monet discovers Dru’s secret artistic side when she finds a sketch in his bag, though she chooses to lie and tell him she believes he is "capable" of running the business despite his father's doubts. Love Triangles and Double Agents Tariq’s personal life is getting as messy as his professional one. He shares a kiss with Lauren at a high-tension dinner with her snobby parents and professors, only to be interrupted by her "cheeseball" boyfriend, Malcolm. Meanwhile, he also shares a moment and a kiss with Diana Tejada when she delivers product to him on campus. Outside the campus bubble, Cooper Saxe is still on the hunt. He enlists his niece, Riley, to go undercover and spy on Tariq at Stansfield. She successfully snaps a photo of Tariq that could spell trouble down the line. Tasha’s Warning From behind bars, Tasha is growing increasingly paranoid. Sensing the danger her son is in, she instructs Tariq to find a gun and protect himself, even pointing him toward one of Ghost and Tommy’s old stash spots. As Tariq balances his Ivy League education with his rising status as a "Prince" of the drug trade, it's clear he's more like his father than he'd ever care to admit. Are you ready to see how He’s already late for his first law school study group
Tariq doesn’t flinch. But when he gets back to his dorm, (Method Man)—his slick, morally flexible attorney—is waiting. Davis drops a bombshell: the feds are building a Rico case using Jabari’s missing laptop. And Jabari had footage of Tariq visiting him the night he died.