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

• Published in: Biotechnology letters 2004-08, Vol.26 (16), p.1265-1268
• Main Authors: Soria, F, Ellenrieder, G, Grasselli, M, Navarro del Canizo, A.A, Cascone, O
• Format: Article
• Language: English
• Subjects: Adsorption; Aspergillus - enzymology; Aspergillus terreus; beta-Glucosidase - biosynthesis; beta-Glucosidase - chemistry; beta-Glucosidase - isolation & purification; Biological and medical sciences; Biological Sciences; Biotechnology; Chemical Fractionation - methods; Chromatography; Chromatography, Affinity - methods; Chromatography, Agarose - methods; Coloring Agents; Dyes; Enzyme Activation; food microbiology; food processing; Fractionation; Fundamental and applied biological sciences. Psychology; Glycoside Hydrolases - biosynthesis; Glycoside Hydrolases - chemistry; Glycoside Hydrolases - isolation & purification; horticultural crops; Hydrogen-Ion Concentration; Multienzyme Complexes - biosynthesis; Multienzyme Complexes - chemistry; Multienzyme Complexes - isolation & purification; Temperature; Triazines
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1023/B:BILE.0000044870.99039.19

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).