Antitumor and HIV-1 Reverse Transcriptase Inhibitory Activities of a Hemagglutinin and a Protease Inhibitor from Mini-Black Soybean
Figure 1
(a) Ion exchange chromatography on a SP-Sepharose column (2.5 cm × 30 cm). Sample: fraction of the mini-black soybean extract adsorbed on Q-Sepharose and subsequently eluted by 0.2 M NaCl in 20 mM Tris-HCl buffer. The dotted slanting line represents the linear NaCl concentration gradient (0–0.5 M) in 100 mM NH4OAc buffer (pH 4.5) used to elute the absorbed proteins from the column. Unabsorbed proteins eluted with 100 mM NH4OAc buffer (pH 4.5) are not shown. Hemagglutinating activity was observed in fraction SP3. Trypsin-inhibitory activity was found in fraction SP1. Fraction size: 13 mL, flow rate: 30 mL/h. Conc. Of NaCl (M- - -), and absorbance at 280 nm (●). The vertical lines in the elution profile denote the pooling of eluate into SP1, SP2, and SP3. (b) Ion exchange chromatography on a DEAE-cellulose column (2.5 cm × 30 cm). Sample: Fraction SP1, the first adsorbed fraction from SP-Sepharose. Adsorbed proteins were eluted from the column by using a linear NaCl concentration gradient (0–0.5 M) in 20 mM Tris-HCl buffer (pH 7.4) represented by the dotted slanting line across the chromatogram. Trypsin-inhibitory activity was observed only in fraction D2. Unabsorbed proteins eluted with 20 mM Tris-HCl buffer (pH 7.4) are not shown. Fraction size: 6 mL, flow rate: 30 mL/h, Conc of NaCl (M- - -), and absorbance at 280 nm (●).
