## 3 Results
### Allergenicity Potential  
The phenomenon wherein a specific antibody recognizes multiple allergens is called allergic cross-reactivity, which provides important information on allergic diseases and their diagnoses [36; 37]. When there is more than 35% identity in a segment of 80 or more amino acids, immunoglobulin E (IgE) cross-reactivity between the newly expressed protein and a known allergen should be considered a possibility [38; 39]. Based on this criterion, a full-length FASTA alignment was done using three different databases to determine the potential allergenicity of the expressed Mut2 protein (Table 7). The results showed that the Mut2 protein has potential allergenicity as evidenced by the % identity with the known allergens on the databases. In the first three databases, two groups of allergens were found to have more than 35% identity with the Mut2 protein, and these were _Lens culinaris_ allergens (56-57%) and _Arachis hypogaea_ allergens (48-49%). According to the Allergen Database for Food Safety, in addition to the previous groups of allergens, _Lupinus angustifolius_ conglutin beta was also found to match the criteria with 56% identity with the Mut protein. In a study by [PERSON] et al. (2011) [11], 55% identity was found in all mutant proteins with _Lens culinaris_ allergen Len C 1.0102. Additionally, based on UniProt and Allergome database queries, sequences matching up to 99.8% were found in species, including _Vigna radiata_, _Vigna angularis_, _Glycine soja_, and _Glycine max_. Bioinformatics analysis showed that the mutated 8S\(\alpha\) globulin can be a potential allergen; however, in vitro and in vivo experiments are still needed to further validate these findings.  
\begin{table}
\begin{tabular}{l c c} \hline \hline
**Database/Allergens** & **\% Identity** & **\(E\)-Value** \\ \hline
**Allermatch [29]** & & \\ _Lens culinaris_ Allergen Len c 1.0102 & & 57.00 & \(1.9\times 10^{-51}\) \\ _Lens culinaris_ Allergen Len c 1.0101 & & 56.40 & \(2.7\times 10^{-51}\) \\ _Arachis hypogaea_ Allergen Ara h 1, clone P17 & & 50.60 & \(4.4\times 10^{-31}\) \\ _Arachis hypogaae_ Allergen Ara h 1, clone P41B & & 49.20 & \(2.8\times 10^{-26}\) \\ _Arachis hypogaaea_ Ara h 1 allergen & & 48.40 & \(1.8\times 10^{-17}\) \\ **AllergenOnline [27]** & & & \\ _Lens culinaris_ Allergen Len c 1.0102 & & 57.00 & \(2.2\times 10^{-60}\) \\ _Lens culinaris_ Allergen Len c 1.0101 & & 56.40 & \(3.2\times 10^{-60}\) \\ _Arachis hypogaaea_ Allergen Ara h 1, clone P17 & & 50.60 & \(1.1\times 10^{-36}\) \\ _Arachis hypogaae_ Allergen Ara h 1, clone P41B & & 49.20 & \(4.5\times 10^{-31}\) \\ _Arachis hypogaaea_ Ara h 1 allergen & & 48.40 & \(8.6\times 10^{-21}\) \\ _Achis hypogaae_ Ara h 1 allergen & & 56.20 & \(6.2\times 10^{-91}\) \\ _Lens culinaris_ Allergen Len c 1.0102 & & 56.80 & \(4.1\times 10^{-64}\) \\ _Lens culinaris_ Allergen Len c 1.0101 & & 55.90 & \(5.5\times 10^{-64}\) \\ _Arachis hypogaae_ Allergen Ara h 1, clone P17 & & 50.60 & \(1.5\times 10^{-38}\) \\ _Arachis hypogaaea_ Allergen Ara h 1, clone P41B & & 49.80 & \(3.5\times 10^{-33}\) \\ _Arachis hypogaaea_ Ara h 1 allergen & & 48.50 & \(4.0\times 10^{-21}\) \\ \hline \hline \end{tabular}
\end{table}
Table 7: Analysis of the allergenic potential of Mut2 8S\(\alpha\) globulin using full-length FASTA alignment.  
Based on the epitope search conducted in ADFS [26] and AllerBase [28], the IIAEK peptide matched the Bos d 5 epitopes (Table 8), which was expected since the source of this peptide was the bovine milk protein, \(\beta\)-lactoglobulin [8]. Previous studies have shown that structural similarity of allergens, specifically their IgE-binding epitopes, plays a critical role in cross-reactivity [38; 39].