## 3 Results
### Identification and Functional Annotation of Differentially Expressed Metabolites
The significant enrichment of metabolites involved in terpenoid backbone biosynthesis was demonstrated (rich factor (RF) value of 1). Differential metabolites involved in photosynthesis also showed significant enrichment, with an RF value equal to 0.65 (Figure 6). Functions related to the biosynthesis of secondary metabolites including lutein (\(\beta\),\(\varepsilon\)-carotene-3,3'-diol), the predominant compound in the 'NuMex LotaLutein' serrano pepper, have been annotated in the KEGG Orthology database for a number of differentially expressed metabolites identified. \begin{table}
\begin{tabular}{l c c c c c c c} \hline \hline \multirow{2}{*}{**Metabolite**} & \multirow{2}{*}{**Class**} & \multicolumn{2}{c}{**Variable**} & \multirow{2}{*}{**Fold**} & \multirow{2}{*}{**log\({}_{2}\)FC**} & \multirow{2}{*}{**KEGG**} & \multirow{2}{*}{**Annotation**} \\  & & & **Imporance in** & & & & **Change** \\  & & & **Projection** & & **(FC)** & & **(KO) Index** & \\ \hline Chlorogenic acid & \multirow{2}{*}{**Class**} & \multirow{2}{*}{**Class**} & \multirow{2}{*}{1.43} & \multirow{2}{*}{20.46} & \multirow{2}{*}{4.35} & KO00940; & \multirow{2}{*}{**C.256**} & \multirow{2}{*}{**Annotation**} \\ (3-O-Caffeoylquinic acid) & & & & & & & & \\  & & & & & & & & \\  & & & & & & & & \\ \hline
3,7-Di-O-methylquercetin & \multirow{2}{*}{**Flavonoids**} & \multirow{2}{*}{1.34} & \multirow{2}{*}{11.21} & \multirow{2}{*}{3.49} & KO00944; & \multirow{2}{*}{**Phenylpropanol biosynthesis;} \\  & & & & & & & & \\  & & & & & & & & \\ \hline Phenol & \multirow{2}{*}{**C.256**} & \multirow{2}{*}{1.42} & \multirow{2}{*}{8.39} & KO009190; & \multirow{2}{*}{**C.256**} & \multirow{2}{*}{**C.256**} \\  & & & & & & & & \\  & & & & & & & & \\  & & & & & & & & \\ ATP; Adenosine & \multirow{2}{*}{**C.257**} & \multirow{2}{*}{**Flavonoids**} & \multirow{2}{*}{1.42} & \multirow{2}{*}{8.39} & KO00908 & \multirow{2}{*}{**C.256**} \\ \(5^{7}\)-Triphosphate & & & & & & & & \\  & & & & & & & & \\  & & & & & & & & \\  & & & & & & & & \\  & & & & & & & & \\  & & & & & & & & \\ \hline
2-Furoic acid & Organic acids & 1.38 & 5.77 & 2.53 & KO01100 & Metabolic pathways \\ \hline Kaempfer-3-O- & \multirow{2}{*}{**Flavonoids**} & \multirow{2}{*}{1.18} & \multirow{2}{*}{5.61} & \multirow{2}{*}{2.49} & KO00944; & \multirow{2}{*}{Flavone and flavonoid biosynthesis;} \\  & & & & & & & & \\  & & & & & & & & \\  & & & & & & & & \\ \hline
2-Oxoheptanedionic acid & Organic acids & 1.37 & 5.55 & 2.47 & KO01100; & \multirow{2}{*}{**C.256**} & \multirow{2}{*}{**C.256**} \\  & & & & & & & & \\  & & & & & & & & \\  & & & & & & & & \\ \hline Kaempfer-3-O- & \multirow{2}{*}{**Flavonoids**} & \multirow{2}{*}{1.31} & \multirow{2}{*}{4.92} & \multirow{2}{*}{2.30} & KO00944 & \multirow{2}{*}{Flavone and flavonoid biosynthesis } \\ (Afzein)(Kaempfer) & & & & & & & & \\ \hline Luteolin-7-O-glucuron & \multirow{2}{*}{**Flavonoids**} & \multirow{2}{*}{1.26} & \multirow{2}{*}{4.76} & \multirow{2}{*}{2.25} & KO00944 & \multirow{2}{*}{Flavone and flavonoid biosynthesis } \\ \(3\)-O-Methylquercetin & \multirow{2}{*}{**Flavonoids**} & \multirow{2}{*}{1.40} & \multirow{2}{*}{4.56} & \multirow{2}{*}{2.19} & KO00944; & \multirow{2}{*}{**Flavone and flavonoid biosynthesis**} \\  & & & & & & & \\  & & & & & & & \\  & & & & & & & \\  & & & & & & & \\ \hline \hline \end{tabular}
\end{table} 
Table 2: Top 10 differentially expressed upregulated metabolites with KEGG database annotation. Figure 5: Summary of the different functions annotated for the differentially expressed metabolites (DEMs) using the KEGG database. Annotations revealed that the majority of the DEMs are involved in metabolic pathways and in the biosynthesis of secondary metabolites and cofactors. Note: A DEM can have multiple annotated functions.