Thirty fungal species grown on Cichorium intybus L. root extract as a sole 
carbon source, were screened for the production of exo-inulinase activities. The 
thermophile Thielavia terrestris NRRL 8126 and mesophile Aspergillus foetidus 
NRRL 337 gave the highest production levels of inulinases I & II at 50 and 24 ºC 
respectively. Yeast extract and peptone were the best nitrogen sources for 
highest production of inulinases I & II at five and seven days of incubation 
respectively. The two inulinases I & II were purified to homogeneity by 
gel-filtration and ion-exchange chromatography with 66.0 and 42.0 fold of 
purification respectively. The optimum temperatures of purified inulinases I & 
II were 75 and 50 ºC respectively. Inulinase I was more thermostable than the 
other one. The optimum pH for activity was found to be 4.5 and 5.5 for 
inulinases I & II respectively. A comparatively lower Michaelis-Menten constant 
(2.15 mg/ml) and higher maximum initial velocity (115 µmol/min/mg of protein) 
for inulinase I on inulin demonstrated the exoinulinase's greater affinity for 
inulin substrate. These findings are significant for its potential industrial 
application. The molecular mass of the inulinases I & II were estimated to be 72 
& 78 kDa respectively by sodium dodecyl sulfate-polyacrylamide gel 
electrophoresis.