## 2 Materials and Methods
### Sample Preparation and Analysis
[PERSON] [24] investigated the influence of the milling degree on the mashing process and found that the increase in the milling degree leads to an increase in the breakdown of high-molecular protein fractions, which consequently leads to an increase in the concentration of their degradation products in word, with the largest increase of FAN concentration already at the very beginning of the mashing process. When using wheat as unmalted raw material (16% substitution for malt), it was found that the concentrations of soluble proteins, HMW, MWM, and LWM are similar to those of the control ports produced from barley malt, and the concentrations of formol N and FAN are even lower [25], while in the case of using wheat malt, the degree of grain modification (as a consequence of the applied malting procedure) has a very significant influence on their concentration in the word [26]. [PERSON] et al. [24] examined the influence of different milling procedures (upward/isothermal) and preparation procedures (milling procedure, grits modification, grits: liquor ratio), from where indicators of cytolysis, proteolysis, and amylolysis were measured \(\beta\)-glucan, FAN and extract. He concluded that malt modification is the most important factor responsible for the breakdown of \(\beta\)-glucan and the release of FAN, while starch modification is responsible for extract yield, and for milling degree, he did not find that it significantly affects the mentioned indicators except in the case of poorly modified malt. From the results in (Table 3), it can be seen that in hard wheat (sample 1), the concentration of total N, medium molecular weight N (MM N), and soluble N increases significantly with the increase of milling degree, and with a significant increase of filtration time. For the attenuation limit (AL), a very clear difference between fine and coarse granulation was established, in such a way that fine granulation has a significantly higher AL compared to coarse in all types of wheat, except for soft, where this difference for AL is the smallest. As for the main indicators of protein breakdown (Kolbach index, soluble proteins, and FAN), no significant difference was found between fine and coarse milling, nor for extract, viscosity, and Hartong 45\({}^{\circ}\), so this significant increase in AL can be attributed to the increase in milling degree. The same trend of a significant increase in AL is also observed for transitional wheats (samples 2 and 3), although with them, with an increase in the milling degree, a significant decrease in MM N and FAN and a slightly less pronounced increase in the filtration time were observed. Compared to barley, wheat has a lower proportion of \(\beta\)-glucan and a higher arabin xylan content, which should result in higher mash viscosity and reduced filtration volume [27; 28]. As a reason for increasing the filtration time and in the case of very good values for the viscosity of wheat word authors [25] state the possibility of forming a protein or protein-polysaccharide gel on the filter, which was also observed when unmalted wheat was used as a substitute for part of the malt in the infusion [25]. When the results for viscosity and Hartong number are observed, the already mentioned negative influence of the \"forcing maturation\" effect caused by the climate, characterized by high temperatures and high humidity towards the end of the growing season, results in a lower enzymatic strength compared to northern European wheats. This effect could be responsible for the lower proportion of soluble in total pentosans also compared to Northern European wheats [29], which also affects the viscosity of word. In the case of soft wheat (sample 4), AL is also significantly higher for fine granulation with a very high, practically unacceptable, filtration time and a significant increase in viscosity for the same. The best values for AL, but also almost all tested parameters, were shown by sample no. 2 (Tika-Taka) with fine grit granulation (\(\alpha\) 0.2 mm).