## 3 Results
### Nitrogen Consumption Differentially Impacts the Production of Volatile Compounds  
The ability to assimilate different nitrogen sources in the brewing word directly impacts the physiology of yeasts during the fermentation process. Simultaneously, the differential consumption of nitrogen sources could affect the aroma and flavor profiles of fermented word [1; 28; 29; 30]. In this context, to determine the nitrogen consumption profiles of the two strains (CL905.1 and CL602.1) at 12 \({}^{\circ}\)C and 20 \({}^{\circ}\)C during fermentation, yeast assimilable nitrogen (YAN) consumption was measured by HPLC after 24, 48, and 168 h of fermentation (Figure 6). In the case of ammonium, that represents 29.02% (Table S4) of the overall YAN in the beer word, \(21.98\%\pm 1.31\%\) of all the available ammonium was consumed by 24 h at 20 \({}^{\circ}\)C, while at 12 \({}^{\circ}\)C, a similar fraction (\(14.82\%\pm 10.50\%\), \(p\)-value \(<0.001\), one-way ANOVA) was reached at 48 h (Figure 6a, Table S4). In this way, when we compared the effect of temperature over the consumption of ammonium in both strains, significant differences were observed only at 24 h (\(p\)-value \(<0.05\), ANOVA) (Figure 6a), while at 168 h, most of the ammonium was already consumed in both strains, suggesting a temperature dependent (and not strain dependent) nitrogen consumption rate.  
Subsequently, we analyzed the consumption of amino acids in word. In this way, we found that their consumption rate was slower compared to ammonium, where both strains exhibited significant differences at 24 h between 12 \({}^{\circ}\)C and 20 \({}^{\circ}\)C (Figure 6a, Table S4). Strain CL905.1 consumed \(82.44\pm 1.95\) mg N/L at 12 \({}^{\circ}\)C, while a higher value was reached (\(125.77\pm 2.13\) mg N/L, \(p\)-value \(<0.001\), ANOVA) when the fermentation temperature was set to 20 \({}^{\circ}\)C. Similarly, CL602.1 showed an increase in amino acid consumption between 12 \({}^{\circ}\)C (91.56 \(\pm\) 8.41 mg N/L) and 20 \({}^{\circ}\)C (130.36 \(\pm\) 3.28 mg N/L,  
Figure 6: Differential yeast assimilable nitrogen (YAN) consumption in CL905.1 and CL602.1 strains at 12 \({}^{\circ}\)C and 20 \({}^{\circ}\)C. (**a**) Ammonium and amino acid consumption kinetics at 24, 48, and 168 h. Blue dots: 12 \({}^{\circ}\)C fermentation; Red dots: 20 \({}^{\circ}\)C fermentation. (*) depict significant differences between temperatures with a \(p\)-value \(<0.001\), one-way ANOVA. (**b**) Main differences in amino Acid consumption between temperatures. Different letters depict significant differences between strains with a \(p\)-value \(<0.05\), one-way ANOVA.  
\(p\)-value \(<\) 0.001, ANOVA) (Table S4), demonstrating differential amino acid consumption depending solely on the fermentation temperature, but not on these genetic backgrounds. Of all the amino acids analyzed in this study (Table S5), solely glutamic acid and aspartic acid significantly differed at 12 \({}^{\circ}\)C (Figure 6b) (\(p\)-value \(<\)0.05, ANOVA) between strains, while for the rest of the amino acids, the differences were due to an increase in fermentation temperature, independently of the strain (Figure 6b). Additionally, we determined that, for both strains, several amino acids, such as aspartic acid, glutamic acid, serine, and threonine, were rapidly consumed in the word (\(>\)60%) at 24 h, demonstrating the preference of _S. eubayanus_ strains for these nitrogen sources (Figure 7, Table S5). On the other hand, amino acids such as histidine, glycine, tyrosine, and cysteine had lower consumption levels (\(<\)30%) at 24 h (low preference) and were efficiently consumed (\(>\)80%) only at 168 h (Table S7).