从发芽蚕豆（阿拉梅达蚕豆变种）中纯化和鉴定一种植酸降解酶

Purification and characterization of a phytate-degrading enzyme from germinated faba beans (Vicia faba Var. Alameda).

作者信息

Greiner R, Muzquiz M, Burbano C, Cuadrado C, Pedrosa M M, Goyoaga C

机构信息

Federal Research Centre for Nutrition, Centre for Molecular Biology, Haid-und-Neu-Strasse 9, D-76131 Karlsruhe, Germany.

出版信息

J Agric Food Chem. 2001 May;49(5):2234-40. doi: 10.1021/jf0100806.

DOI:10.1021/jf0100806

PMID:11368582

Abstract

A phytate-degrading enzyme was purified approximately 2190-fold from germinated 4-day-old faba bean seedlings to apparent homogeneity with a recovery of 6% referred to the phytase activity in the crude extract. It behaves as a monomeric protein of a molecular mass of approximately 65 kDa. The phytate-degrading enzyme belongs to the acidic phytases. It exhibits a single pH optimum at 5.0. Optimal temperature for the degradation of sodium phytate is 50 degrees C. Kinetic parameters for the hydrolysis of sodium phytate are K(M) = 148 micromol L(-1) and k(cat) = 704 s(-1) at 35 degrees C and pH 5.0. The faba bean phytase exhibits a broad affinity for various phosphorylated compounds and hydrolyzes phytate in a stepwise manner. The first hydrolysis product was identified as D/L-myo-inositol(1,2,3,4,5)pentakisphosphate.

摘要

从萌发4天的蚕豆幼苗中纯化出一种植酸降解酶，纯化倍数约为2190倍，达到表观均一性，相对于粗提物中的植酸酶活性，回收率为6%。它表现为一种分子量约为65 kDa的单体蛋白。该植酸降解酶属于酸性植酸酶。其最适pH值为5.0。降解植酸钠的最适温度为50℃。在35℃和pH 5.0条件下，植酸钠水解的动力学参数为K(M)=148 μmol L(-1)，k(cat)=704 s(-1)。蚕豆植酸酶对各种磷酸化化合物具有广泛的亲和力，并以逐步方式水解植酸。第一个水解产物被鉴定为D/L-肌醇(1,2,3,4,5)五磷酸。