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/).
In Thailand, ladyboys are more visible and socially accepted than transgender individuals in many other parts of the world. They can be found working in all sectors, from retail to entertainment, and even have their own dedicated restrooms in some public establishments. This relative tolerance stems from deep-rooted cultural and religious beliefs, where the concept of karma and reincarnation can frame a Kathoey identity not as a defect or sin, but as the result of past-life actions.
: Beyond social media, she has explored diverse interests, including winning a boxing match in Phuket to demonstrate resilience and strength. Cultural Context
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The broader term "ladyboy" is a colloquially adapted English translation of the Thai term Kathoey . Historically localized to Southeast Asian culture, the term has transitioned into a globalized label within online media and adult entertainment industries.
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| Platform | Revenue Stream | Avg. Monthly Income (USD) per Performer* | |----------|----------------|------------------------------------------| | YouTube (AdSense + Super Chat) | Advertising + Live‑chat donations | 3,200 | | TikTok Creator Fund | Algorithmic payouts | 2,900 | | Patreon (“Ultra‑Luxe”) | Subscription tiers (US$10–30) | 5,400 | | Twitch
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Sawasdee Lydia’s rise highlights how digital platforms like Instagram and OnlyFans provide a space for transgender individuals to cultivate their own brand, gain financial independence, and connect with a global audience. Her, narrative, as explored in discussions like those found on Apple Podcasts , makes her one of the most prominent, albeit sometimes misunderstood, voices in the modern Thai digital scene.
She is active on Instagram as @lydialadyboy and Facebook as SawasdeeLydia , where she frequently posts about beach life, fashion, and personal reflections.
Lidia is recognized for her engaging personality and her ability to inspire others to live on their own terms. In the context of her work with , she represents a broader movement of transgender women in Thailand and beyond who utilize digital platforms to reclaim their narratives. Key aspects of her digital career include:
In Thailand, ladyboys are more visible and socially accepted than transgender individuals in many other parts of the world. They can be found working in all sectors, from retail to entertainment, and even have their own dedicated restrooms in some public establishments. This relative tolerance stems from deep-rooted cultural and religious beliefs, where the concept of karma and reincarnation can frame a Kathoey identity not as a defect or sin, but as the result of past-life actions.
: Beyond social media, she has explored diverse interests, including winning a boxing match in Phuket to demonstrate resilience and strength. Cultural Context
Do you require information on when navigating file-sharing sites?
While "extremeladyboys lidia" might not refer to a specific webpage, the name "Lydia" is prominently linked to the community this platform serves. The individual at the heart of the search term is a successful content creator and a well-known transgender woman (or "ladyboy," a common term in Thailand for transgender women) living in Phuket, Thailand. extremeladyboys lidia
If you're looking for information on a specific topic or individual, I can certainly try to help with that. For instance, if you're interested in learning more about a particular subject or person, I can provide you with general information, suggest reputable sources, or even help you craft a well-researched write-up.
The broader term "ladyboy" is a colloquially adapted English translation of the Thai term Kathoey . Historically localized to Southeast Asian culture, the term has transitioned into a globalized label within online media and adult entertainment industries.
While many transgender women work in this industry, Lydia stands out for being named "one of Thailand's most prominent ladyboys". The table below highlights key differences between her and others in the field: In Thailand, ladyboys are more visible and socially
Because this topic falls under adult entertainment and lacks mainstream biographical, cultural, or historical documentation, creating a long-form factual article is not possible.
| Platform | Revenue Stream | Avg. Monthly Income (USD) per Performer* | |----------|----------------|------------------------------------------| | YouTube (AdSense + Super Chat) | Advertising + Live‑chat donations | 3,200 | | TikTok Creator Fund | Algorithmic payouts | 2,900 | | Patreon (“Ultra‑Luxe”) | Subscription tiers (US$10–30) | 5,400 | | Twitch
Lidia is known for her bold personality and her commitment to authenticity. She founded the Extremeladyboys channel as a way to share her life and perspective with a global audience, moving beyond traditional stereotypes. Her content often features: : Beyond social media, she has explored diverse
I can’t help with requests that sexualize or exploit real people or groups. If you’d like, I can:
Sawasdee Lydia’s rise highlights how digital platforms like Instagram and OnlyFans provide a space for transgender individuals to cultivate their own brand, gain financial independence, and connect with a global audience. Her, narrative, as explored in discussions like those found on Apple Podcasts , makes her one of the most prominent, albeit sometimes misunderstood, voices in the modern Thai digital scene.
She is active on Instagram as @lydialadyboy and Facebook as SawasdeeLydia , where she frequently posts about beach life, fashion, and personal reflections.
Lidia is recognized for her engaging personality and her ability to inspire others to live on their own terms. In the context of her work with , she represents a broader movement of transgender women in Thailand and beyond who utilize digital platforms to reclaim their narratives. Key aspects of her digital career include: