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8138783 
Journal Article 
Partial purification and characterization of exoinulinase from Kluyveromyces marxianus YS-1 for preparation of high-fructose syrup 
Singh, RS; Dhaliwal, R; Puri, M 
2007 
Yes 
Journal of Microbiology and Biotechnology
ISSN: 1017-7825
EISSN: 1738-8872 
17 
733-738 
English 
An extracellular exoinulinase (2,1-beta-D fructan fructanohydrolase, EC 3.2.1.7), which catalyzes the hydrolysis of inulin into fructose and glucose, was purified 23.5-fold by ethanol precipitation, followed by Sephadex G-100 gel permeation from a cell-free extract of Kluyveromyces marxianus YS-1. The partially purified enzyme exhibited considerable activity between pH 5 to 6, with an optimum pH of 5.5, while it remained stable (100%) for 3 h at the optimum temperature of 50 degrees C. Mn2+ and Ca2+ produced a 2.4-fold and 1.2-fold enhancement in enzyme activity, whereas Hg2+ and Ag2+ completely inhibited the inulinase. A preparation of the partially purified enzyme effectively hydrolyzed inulin, sucrose, and raffinose, yet no activity was found with starch, lactose, and maltose. The enzyme preparation was then successfully used to hydrolyze pure inulin and raw inulin from Asparagus racemosus for the preparation of a high-fructose syrup. In a batch system, the exoinulinase hydrolyzed 84.8% of the pure inulin and 86.7% of the raw Asparagus racemosus inulin, where fructose represented 43.6 mg/ml and 41.3 mg/ml, respectively. 
Asparagus Plant/chemistry; Cations, Divalent/pharmacology; Chemical Fractionation; Chromatography, Gel; Dextrans; Enzyme Activators/pharmacology; Enzyme Inhibitors/pharmacology; Enzyme Stability; Fructose/metabolism; Glycoside Hydrolases/isolation & purification/metabolism; Hydrogen-Ion Concentration; Inulin/isolation & purification/metabolism; Kluyveromyces/enzymology; Liliaceae; Metals/pharmacology; Orchidaceae; Plant Extracts/metabolism; Raffinose/metabolism; Substrate Specificity; Sucrose/metabolism; Temperature; Time Factors