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 alpha-L-rhamnosidase and two beta-D-glucosidases (beta G1 and beta G2) 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 beta-D-glucosidases was from pH 4 to 6 and at 65 degrees C. Both glucosidases were active on p -nitrophenol glucoside and prunin with respective Km values of 1.9 mm and 1.6 mm for beta G1 and 2.1 mm and 0.25 mm for beta G2. Only beta G1 hydrolysed cellobiose (Km = 5.7 mm).
Chemical Industry Research Institute (INIQUI), National University of Salta - CONICET, Buenos Aires 177 - 4400 Salta, Argentina.
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Soria
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Ellenrieder
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Grasselli
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Navarro del Cañizo
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Cascone
D000327:Adsorption; D001230:Aspergillus / Q000201:enzymology*; D005591:Chemical Fractionation / Q000379:methods*; D002846:Chromatography, Affinity / Q000379:methods*; D002847:Chromatography, Agarose / Q000379:methods*; D004396:Coloring Agents; D004789:Enzyme Activation; D006026:Glycoside Hydrolases / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D009097:Multienzyme Complexes / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification*; D013696:Temperature; D014227:Triazines; D001617:beta-Glucosidase / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification*
D004396:Coloring Agents; D009097:Multienzyme Complexes; D014227:Triazines; D006026:Glycoside Hydrolases; C039827:naringinase; D001617:beta-Glucosidase