IMMOBILIZATION OF ALKALINE-PHOSPHATASE BY SORPTION ON NA-SEPIOLITE

被引:27
作者
CARRASCO, MS
RAD, JC
GONZALEZCARCEDO, S
机构
关键词
ADSORPTION; ALKALINE PHOSPHATASE; CLAYS; ENZYME IMMOBILIZATION; ENZYME KINETICS; SOIL ENZYMES; SEPIOLITE;
D O I
10.1016/0960-8524(94)00115-H
中图分类号
S2 [农业工程];
学科分类号
0828 ;
摘要
Alkaline phosphatase from bovine intestinal mucous membrane was immobilized by sorption on Na-sepiolite in order to obtain an enzyme-clay complex for soil application. Optimum conditions for immobilization were: phosphate buffer 0.1 M at pH 7, 2 h of magnetic stirring at 4 degrees C, 0.25 mg ml(-1) for the concentration of enzyme and an enzyme/clay ratio of IO ml g(-1). The adsorption isotherm obeyed the Langmuir equation (r(2) 0.996 at P<0.001), reaching a maximum amount of enzyme retention. Apparent K-m values were 18.32 mM for the immobilized enzyme and 2.36 mM for the enzyme in aqueous solution; optimum pH (10.5) did not change as a consequence of immobilization; a first-order kinetic was obtained for incubation times of less than I h for both enzymes and the immobilized enzyme showed a lower incubation temperature optimum. The immobilized phosphatase showed less stability than the soluble enzyme in terms of storage at 30 degrees C, thermal stability and resistance to proteolytic attack.
引用
收藏
页码:175 / 181
页数:7
相关论文
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