Zymolyase digestion
A glucan sample (2 mg) suspended in acetate buffer
(50 mM, pH 6.0) was mixed with 100 μg/ml of zymolyase
100T (Seikagaku, Tokyo, Japan). After incubation
overnight at 45◦C, the reaction mixture was boiled
for 3 min to inactivate the enzyme. The resulting solution
was mixed with ethanol (1:4) and soluble and insoluble
fractions prepared. The ratio was monitored using the
phenol-H2SO4 method.