## 3 Results and Discussion
### Intracellular Polysaccharide Synthesis and Composition  
The data given in Table 2a-c demonstrate the highest values of cellular carbohydrates in both absolute (g/L) and relative (%, w/w) values. IPS values (in g/L and %, w/w) constantly increased as a function of the fermentation time for _Morchella_ species when they were cultivated on S-20 (Figure 1a), both in static and agitated cultures and agitation seemed to have a positive impact. Even so, the kinetic profiles of produced IPS absolute values on S-25, M-20, M-25 (Figure 1b-d, respectively) showed that there was a notable difference between agitated and static cultures, as IPS constantly increased until the end of the fermentation on agitated flasks, whereas a reduction in the IPS (g/L) values was detected towards the end of static cultures. This fact showed that agitation may contribute to the adequate supply of nutrients to cells and facilitated the removal of gases and other by-products of catabolism from the microenvironment of cells until the end of the fermentation [57]. Moreover, agitation also had positive or no impact upon the biosynthesis of IPS on these substrates, depending on species and kind of substrate. Most of the highest relative IPS values (\(\%\), w/w) were observed in the intermediate days of cultivation. Among species, _M. conica_ achieved the highest values in all nutrition media (4.8 g/L; M-20, 4.6 g/L; S-20 and S-25, 3.1 g/L; M-25), corresponding to quantities up to 21% and even reached 51.8%, w/w of IPS in dry mycelial mass-produced. The most suitable medium for fungal IPS production was molasses substrate M-20, but when the C/N ratio increased (M-25) there was a significant decrease in IPS production (in both g/L and \(\%\), w/w values). [PERSON] et al. [58] showed that an increase in the (C/N) ratio of the medium caused the reduction of IPS concentration in the media. In the current investigation, the results concerning the high amounts of IPS achieved reflect the high ability of _Morchella_ polysaccharide synthesis and the potential to be used as functional foods. The concentration of IPS\({}_{\text{max}}\) produced by _Morchella_ species in both  
Figure 1: Production rate of intracellular polysaccharides (IPS) in absolute (g/L) values during the cultivation of _Morchella rotunda_, _M. vulgaris_ and _M. conica_ in media containing sucrose (S-20; (**a**), S-25; (**b**)) and molasses (M-20; (**c**), M-25; (**d**)) in static and agitated (120 \(\pm\) 5 rpm) flask liquid cultures of C/N molar ratio 20 and 25.  
agitated and static- flask cultures (2.0-4.6 g/L and \(\sim\)28-44%; S-20, 3.3-4.8 g/L and \(\sim\)20-35%; M-20, 1.8-4.6 g/L and \(\sim\)26-52%; S-25, 1.5-3.1 g/L and \(\sim\)17-33%; M-25) may be compared with those reported in the literature; the yellow morel _M. esculenta_ produced IPS\({}_{\text{max}}\) quantities of \(\sim\)3.8 g/L in glucose-based static and shake-flask cultures proving that they were not significantly affected by agitation [29]. This is very interesting as mushroom polysaccharides. Also, _G. applanatum_ in shake-flasks sucrose-based cultures presented IPS\({}_{\text{max}}\) concentration of 1.5% [58], while _Cordyceps prunosa_ on media composed of molasses in shake-flasks produced IPS\({}_{\text{max}}\) quantities of \(\sim\)5 g/L [59]. In contrast, when [PERSON] et al. [17] cultivated _V. volvacen_ in a glucose-based medium under agitation, the amount of IPS presented a three-fold decrease by the end of cultivation and the highest value was 5.49 g/L (C/N = 20). The carbohydrate composition of IPS depends on the growth medium and it could be changed by manipulating submerged culture's factors [17]. So, in this study, not only the medium used, but also different species, the impact of agitation and the influence of initial sugars' concentration on the composition of IPS were investigated.