Chromatography on RP-HPLC column and SDS–PAGE (15%) of SAL4. (A) RP-HPLC on a C-18 column, elution was performed at room temperature within 40 min using a gradient from 0 to 80% acetonitrile at a flow rate of 1 mL/min. The gradient is indicated by the dotted line. The absorbance was measured at 280 nm. AU: Arbitrary Units. (B) 15%-SDS–PAGE of pure SAL4. Lane 1, molecular mass markers (Pharmacia); lane 2, 7 μg of purified SAL4 eluted from RP-HPLC. (C) Alignment of the amino acid sequences of the mature forms of SAL4 (Present study, KF67886), SAL3 (Horchani et al., 2009), SSL (Sayari et al., 2001), SEL3 (Rosenstein et al., 2000) and SHL (Tiesinga et al., 2007).