Fig. 4

Thin-layer chromatography of the products of β-galactosidase and α-galactosidase activities from G. dipsosauri strain DD1. Small samples (2 μl) were spotted on sheets of Merck Silica Gel F60 and developed in a solvent of n-butanol-n-propanol-ethanol-water (20:30:30:20). The spots were identified by spraying with an orcinol-sulfuric acid mixture and heating at 60 ° °C. Panel (a) shows the results with a 10% (w/v) lactose substrate solution. The samples were as follows: (1) lactose + P80 fraction; (2) lactose + desalted P80 fraction; (3) lactose + MacroPrep DEAE pool containing β-galactosidase I; (4) lactose + MacroPrep DEAE pool containing β-galactosidase II; (5) Sephadex G-200 fraction 16 from MacroPrep DEAE pool containing β-galactosidase I; (6) Sephadex G-200 fraction 21 from the MacroPrep DEAE pool containing β-galactosidase II; (7) 2% (w/v) d-glucose standard; (8) 2% (w/v) d-galactose standard; and (9) 2% (w/v) lactose standard. Panel (b) shows the results with 2% (w/v) d-melibiose or d-raffinose substrate solutions. The samples were as follow: (1) 2% (w/v) d-melibiose standard; (2) melibiose + Sephadex G-200 fraction 22 from the MacroPrep-DEAE pool enriched in α-galactosidase; (3) melibiose + Sephadex G-200 fraction 16 from MacroPrep DEAE pool containing β-galactosidase I; (4) melibiose + Sephadex G-200 fraction 21 from the MacroPrep DEAE pool containing α-galactosidase and β-galactosidase II; (5) 2% d-raffinose standard; (6) raffinose + Sephadex G-200 fraction 22 from the MacroPrep-DEAE pool containing α-galactosidase; (7) raffinose + Sephadex G-200 fraction 16 from the MacroPrep-DEAE pool containing β-galactosidase I; (8) 2% (w/v) d-galactose standard; and (9) 2% (w/v) sucrose standard. The d-glucose standard moved to the same position as sucrose but had a brownish color after staining rather than a purplish color