The supernatant was collected after
centrifugation (5 min, 7000 g), and the residues were washed once
with water (4mL), centrifuged and then combined with the previous
extract. The aqueous extract was applied to a DEAE-Sephadex A-25
(40 mg) column (pyridine acetate form) (GE Healthcare, Piscataway,
NJ).