Figure 1. Screening and purifying of the softener.

(A) The result of the mechanical tests on the Triton-treated dermis of S. chloronotus. In this and Figs 3 and 4 the ordinate is relative stiffness normalized by the peak value in the steady phase of the respective experiment. The application of the crude extract (upward arrow) caused a decrease in stiffness. (B) Anion-exchange chromatography on a Mono-Q column of an extract of sea cucumber dermis. The solid line shows the absorbance at 280 nm; the dotted line shows NaCl concentration. In B-D the fraction marked with a circle had softening activity and that with a cross showed no softening activity, and a horizontal bar indicates the fractions processed for further screening. The left-hand-side ordinate of graphs B-D is given in relative values in which the absorbance of the purified softener, the peak value in panel D, was taken as unity. (C) Gel-filtration chromatography on the two serially-attached Superose 12 columns of the active fractions from the Mono-Q column. (D) Gel-filtration chromatography on the Superdex-peptide column of the active fractions from the Superose 12 columns. (E) The result of SDS-PAGE of the active fraction from Superdex peptide column (left lane). The right lane is molecular weight markers.