Purification and some important characters of extracellular inulinase of Alternaria alternata (Fr.) Keissler.

Protein precipitate of cell-free dialysate of extracellular inulinase (2,1-beta-fructan fructanohydrolase, EC 3.2.1.7) of A. alternata was maximally obtained by methanol. Such protein was fractionated by using 2-step column chromatography on Sephadex G150 and DEAE-cellulose. The partially purified enzyme had activity of 81 x 10(3) U/mg protein, with a yield of 69% of the original activity and the fold of purification was 62. Optimum temperature and pH for the activity of the purified enzyme were found to be 55 degrees C and 4.5, respectively. The enzyme was found to be stable up to 55 degrees C and in pH range of 4 to 5. Ba2+ and Ca2+ were found to stimulate the enzyme activity while Cu2+, Fe3+, Hg2+, and iodoacetate were recorded as strong inhibitors. T(1/2) of the enzyme was estimated to be two weeks and its apparent Km was calculated to be 0.066 M. The enzyme recorded hydrolyzing activity against sucrose and raffinose recording I/S ratio of 0.50. Molecular mass of the enzyme preparation was estimated by gel filtration and found to be 115 +/- 5 kDa.