# Marine microalgae Biomolecules and Their Adhesion Capacity to _Salmonella enterica_ sv. Typhimurium
## 3 Results
### Extraction and Characterization of Polysaccharides from P. gyrans and P. lutheri
Furthermore, the HSQC spectrum (Figure 5a) contained correlations at 3.87;3.46/64.7, 5.14/69.7, and 4.35;4.15/62.0, which were attributed, respectively, to H-1;H-1'/C-1, H-2/C-2, and H-3;H-3'/C-3 of linked glycerol (Gly), indicating the presence of the glycoside sulfoquinovosylglycerol (SQG). Moreover, the anomeric region contained two low-intensity correlations at 4.14/103.5 ppm (\(J_{\rm H1,H2}\) 7.1 Hz) and 4.72/99.0 ppm (\(J_{\rm H1,H2}\) 3.3 Hz), which were attributed to \(\beta\)-Gal\(p\) and \(\alpha\)-Gal\(p\) units, respectively, suggesting the presence of a second glycoside in this fraction, namely digalacosylglycerol (DGG) [42; 43; 44]. The CH\({}_{3}\) correlations at 0.86/13.3 ppm, CH\({}_{2}\) at 1.20/21.1 to 2.3/36.6 ppm with a predominant one at 1.25/28.5 ppm (inverted resonances in the \({}^{13}\)C-DEPT spectrum, Figure 5c'), and carboxylic signals at 171-175 ppm in the \({}^{13}\)C NMR spectra (Figure 5c), as well as double-bond correlations from 5.05/119.8 to 5.50/137.3 ppm, indicated the presence of unsaturated free fatty acid chains [45]. \begin{table}
\begin{tabular}{c c c c c c c c c} \hline \hline \multirow{2}{*}{**Fraction**} & \multirow{2}{*}{**Glycosyl Residues**} & \multicolumn{8}{c}{**Chemical Shifts \({}^{\rm a}\)**} \\ \cline{3-10}  & & & **H-1/C-1** & **H-2/C-2** & **H-3/C-3** & **H-4/C-4** & **H-5/C-5** & **H-6;H-6’/C-6** \\ \hline GW-p & \(\rightarrow\)3)-\(\beta\)-d-Glc\(p\)-(1\(\rightarrow\) & G\({}_{\rm L}\) & 4.53/102.4 & 3.32/72.3 & 3.49/85.6 & 3.28/67.9 & 3.28/75.8 & 3.72; 3.48/60.4 \\ \hline \multirow{8}{*}{GW-s} & \(\rightarrow\)3)-\(\alpha\)-p-Glc\(p\) & G\({}_{\rm\alpha}\) & 5.00/91.5 & nd & nd & nd & nd & nd \\  & \(\rightarrow\)3)-\(\beta\)-d-Glc\(p\) & G\({}_{\rm\beta}\) & 4.39/96.1 & nd & nd & nd & nd & nd \\  & \(\beta\)-d-Glc\(p\)-(1\(\rightarrow\)3 & G\({}_{\rm 3t}\) & 4.38/103.5 & 3.11/73.5 & 3.24/75.6 & 3.12/69.7 & 3.12/75.8 & 3.72; 3.47/60.7 \\ \cline{1-1}  & \(\beta\)-d-Glc\(p\)-(1\(\rightarrow\)6 & G\({}_{\rm 6t}\) & 4.23/102.5 & 3.05/73.3 & 3.23/75.6 & 3.13/69.7 & 3.19/nd & 3.71; 3.49/60.7 \\ \cline{1-1}  & \(\rightarrow\)3)-\(\beta\)-d-Glc\(p\)-(1\(\rightarrow\) & G\({}_{\rm A}\) & 4.49/102.4 & 3.32/72.2 & 3.46/85.6 & 3.26/67.9 & 3.29/75.8 & 3.72; 3.48/60.4 \\ \cline{1-1}  & \(\rightarrow\)6)-\(\beta\)-d-Glc\(p\)-(1\(\rightarrow\) & G\({}_{\rm B}\) & 4.28/102.6 & 3.05/73.3 & 3.23/75.8 & 3.15/69.7 & 3.34/74.4 & 4.00; 3.61/67.9 \\ \cline{1-1}  & \(\rightarrow\)3,6)-\(\beta\)-d-Glc\(p\)-(1\(\rightarrow\) & G\({}_{\rm C}\) & 4.36/102.4 & 3.25/72.0 & 3.40/87.0 & 3.33/67.8 & 3.42/74.4 & 4.00; 3.66/67.9 \\ \hline \hline \end{tabular}
\end{table} 
Table 4: \({}^{1}\)H and \({}^{13}\)C NMR chemical shifts of GW-p and GW-s.  
Additional to glycosides and free fatty acid signals, the \({}^{1}\)H and \({}^{13}\)C NMR spectra of LA-p (Figure 5) showed protein signals in accordance with the high protein content (49.1%) observed in the chemical analyses. Overlapping was expected for some of the protein and free fatty acid resonances. The \({}^{1}\)H NMR spectrum (Figure 5b) showed signals in the region of the backbone amide protons (10.0-7.0 ppm), aromatic and side-chain amide protons (8.0-6.0 ppm), \(\alpha\)-proton (5.5-3.5 ppm), methine, methylene (3.5-1.0 ppm), and methyl protons (2.0-0 ppm) of the aliphatic side chains [46]. The \({}^{13}\)C NMR spectrum (Figure 5c) showed signals corresponding to the carbonyl carbon of the main chain and carboxylic groups of the acidic amino acids side chain (180-165 ppm), aromatic amino acids (140-115 ppm), \(\alpha\) and \(\beta\) carbons (65-45 ppm and 45-25 ppm, respectively), and methylene and methyl groups of the aliphatic side chains (25-10 ppm) [47]. Figure 5: NMR analyses of the LA-p fraction. \({}^{1}\)H-\({}^{13}\)C HSQC (**a**), \({}^{1}\)H NMR (**b**), \({}^{13}\)C NMR (**c**), and partial \({}^{13}\)C-DEPT-135 spectra (**c**) (Me\({}_{2}\)SO-\(d_{6}\), 70 \({}^{\circ}\)C). DG, SQ, and Gly correspond to the glycosyl moiety of digalacacosylglycerol and sulfoquinovosylglycerol, and the linked glycerol units, respectively.