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
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When Nandan returns for good, he struggles to "match the wavelength" of his teenage daughter, leading to various misunderstandings and household squabbles. The Resolution:
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Juliet (Meera Jasmine) and her daughter Aparna (Devika Sanjay) have lived a carefree life in Kerala while the father, Nandan (Jayaram), worked as a mechanic in the Gulf. The Conflict:
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The story centers on (Jayaram), an expat who returns to Kerala after 15 years in the Gulf to start an organic business. While his wife Juliet (Meera Jasmine) and daughter Aparna (Devika Sanjay) have developed a deep, friend-like bond in his absence, Nandan struggles to find common ground with his 17-year-old daughter. His traditional mindset often clashes with Aparna’s modern lifestyle, creating a relatable exploration of the generation gap. Cast and Crew Director: Sathyan Anthikad Writer: Iqbal Kuttippuram Main Cast: Jayaram as Nandan Meera Jasmine as Juliet Devika Sanjay as Aparna Naslen Gafoor as Rohit Music: Vishnu Vijay (Songs) and Rahul Raj (Score) Critical Reception Makal (2022)
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Makal is a gentle reminder of the importance of family, patience, and love. It doesn't offer earth-shattering twists, but rather a warm embrace of a story that feels close to home. If you are looking for a heartwarming Malayalam movie to watch over the weekend, searching for will lead you to this endearing story of a father trying to understand his daughter.
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It’s a relatable story about the changing dynamics within a family, the challenges of parenting a teenager, and the generation gap, all presented in the director's trademark feel-good, realistic style. The film stars seasoned actors .
This post discusses a website/term often associated with pirated streaming. It does not encourage illegal activity; the information below is for awareness, safety, and legal alternatives.
When Nandan returns for good, he struggles to "match the wavelength" of his teenage daughter, leading to various misunderstandings and household squabbles. The Resolution:
When users search for "gomovies malayalam makal," they are often directed toward illegal streaming sites that carry severe malware risks, invasive popup ads, and low-quality video rips. Instead of using unsafe third-party networks, you can access the film through authorized distributors: gomovies malayalam makal
Juliet (Meera Jasmine) and her daughter Aparna (Devika Sanjay) have lived a carefree life in Kerala while the father, Nandan (Jayaram), worked as a mechanic in the Gulf. The Conflict:
While international streaming sites like GoMovies operate in a gray area,cinephiles highly recommend supporting the creators by accessing Malayalam cinema through authorized, high-quality platforms. If you want to experience the vibrant colors of the village, the soulful music scored by Vishnu Vijay, and the crisp dialogues without interruptions, official viewing ensures the best cinematic experience. This post discusses a website/term often associated with
The story centers on (Jayaram), an expat who returns to Kerala after 15 years in the Gulf to start an organic business. While his wife Juliet (Meera Jasmine) and daughter Aparna (Devika Sanjay) have developed a deep, friend-like bond in his absence, Nandan struggles to find common ground with his 17-year-old daughter. His traditional mindset often clashes with Aparna’s modern lifestyle, creating a relatable exploration of the generation gap. Cast and Crew Director: Sathyan Anthikad Writer: Iqbal Kuttippuram Main Cast: Jayaram as Nandan Meera Jasmine as Juliet Devika Sanjay as Aparna Naslen Gafoor as Rohit Music: Vishnu Vijay (Songs) and Rahul Raj (Score) Critical Reception Makal (2022)
The movie garnered massive initial buzz for several structural reasons: The Resolution: When users search for "gomovies malayalam
Makal is a gentle reminder of the importance of family, patience, and love. It doesn't offer earth-shattering twists, but rather a warm embrace of a story that feels close to home. If you are looking for a heartwarming Malayalam movie to watch over the weekend, searching for will lead you to this endearing story of a father trying to understand his daughter.
There are several benefits to using Gomovies Malayalam Makal, including:
It’s a relatable story about the changing dynamics within a family, the challenges of parenting a teenager, and the generation gap, all presented in the director's trademark feel-good, realistic style. The film stars seasoned actors .