Fractionation of the naringinase complex from Aspergillus terreus by dye affinity chromatography

Affinity chromatography with immobilised triazine dyes was used to separate the main enzymes present in the naringinase complex produced by Aspergillus terreus CECT 2663. One α-L-rhamnosidase and two β-D-glucosidases (βG1 and βG2) were separated by a simple two-step procedure involving chromatography with Red HE-3B immobilised on Sepharose 4B first at pH 5.5 and then at pH 4.7. Maximum activity of the β-D-glucosidases was from pH 4 to 6 and at 65 °C. Both glucosidases were active on p-nitrophenol glucoside and prunin with respective Km values of 1.9 mm and 1.6 mm for βG1 and 2.1 mm and 0.25 mm for βG2. Only βG1 hydrolysed cellobiose (Km = 5.7 mm).