About 10 mg of dry cell wall mass was used for polysaccharide extraction, in accordance with François [32 (link)]. After extraction, 1 mL from the final preparation was concentrated 10× by lyophilization. The sugars were analyzed using HPLC (Young Lin YL9100 series system) by a refractive index (RI) detector at 40 °C. The samples were loaded in the REZEX ROA (Phenomenex, Torrance, CA, USA) column (300 × 7.8 mm) at 85 °C and eluted with 0.05 M sulfuric acid at a flow rate of 1.5 mL/min [33 (link)]. The saccharides were quantified based on a standard curve (5–100 mM) using HPLC standards from Sigma-Aldrich (St. Louis, MO, USA): Tetraacetylchitotetraose (Chit4), triacetylchitotriose (Chit3), diacetyl-β-D-chitobiose (Chit2), N-Acetil-β-D-glucosamina (nag), and glucose (gluc).
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