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CHARACTERIZATION OF PHYTASE ACTIVITY IN LUPIN SEED
Author(s) -
SILVA L.G.,
TRUGO L.C.
Publication year - 1996
Publication title -
journal of food biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.507
H-Index - 47
eISSN - 1745-4514
pISSN - 0145-8884
DOI - 10.1111/j.1745-4514.1996.tb00559.x
Subject(s) - phytase , phytic acid , gel permeation chromatography , chromatography , chemistry , fractionation , acid phosphatase , hydrolysis , substrate (aquarium) , high performance liquid chromatography , ammonium , biochemistry , enzyme , biology , organic chemistry , ecology , polymer
Lupin phytase (myo‐inositol hexaphosphate phosphohydrolase) and the degradation of its substrate phytic acid were studied during seed germination. Phytase and acid phosphatase activities were detected in nongerminated seeds and increased from days 1 to 8 of germination, while the concentration of phytic acid decreased during the same period. Phytase was extracted with 2% CaCl 2 solution and partially purified by ammonium sulfate fractionation and HPLC‐gel permeation chromatography. Substrate specificites were determined using pnitrophenylphosphate or phytic acid. The pH and temperature optima for the fraction obtained by gel permeation were 5.0 and 50C respectively, while the Km and Vmaxvalues were 0.33 mM and 0.13 μmol·mL −1 ·min −1 , respectively. It was not possible to separate phytase from phosphatase by use of gel permeation chromatography due to similarities in apparent molecular mass, however resolution of the two enzymes was achieved by DEAE‐cellulose chromatography .