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Characterization of an exoinulinase produced by Aspergillus terreus CCT 4083 grown on sugar cane bagasse
(Journal of Agricultural and Food Chemistry, 2010-02-07) Coitinho, Juliana B.; Guimarães, Valéria M.; Almeida, Maíra N. de; Falkoski, Daniel L.; Queiróz, José H. de; Rezende, Sebastião T. de
Exoinulinase (β- D -fructan fructohydrolase, EC 3.2.1.80) secreted by Aspergillus terreus CCT4083 was obtained using sugar cane bagasse, an agroindustrial residue, as a carbon source. It was further purified from the supernatant culture in a rapid procedure. The enzyme presented 57 kDa on SDS-PAGE and 56 kDa on gel filtration chromatography. Inulin was hydrolyzed by the purified enzyme, yielding D -fructose as the main product. This enzyme showed maximum activity at pH 4.0 and 60 °C and maintained more than 90 and 75% of its original activity at 40 and 50 °C, respectively, after 3.5 h of preincubation. The K M values for inulin, sucrose, and raffinose were 11, 4.20, and 27.89 mM, respectively, and D -fructose was a competitive inhibitor (K i = 47.55 mM). The activation energies for sucrose, raffinose, and inulin were 10.4, 5.61, and 4.44 kcal/mol, respectively. The characteristics of A. terreus exoinulinase were compared to those of inulinases isolated from other organisms. The exoinulinase traits presented especially good thermostability and the ability to produce pure D -fructose, suggesting its application to the production of high-fructose syrup.