Shutov A D, Uen N T, Fridman S A, Vaintraub I A
Biokhimiia. 1975 May-Jun;40(3):553-8.
Hydrolysis of L-phenylalanyl-p-nitroanilide (PPA) and glycyl-p-nitroanilide by extracts from resting vetch seeds was shown to be the effect of two different arylamidases. One of them, PPAase, was 2000-fold purified on hydroxylapatide, DEAE-cellulose and by gel filtration through Sephadex G-100. The preparation obtained was nearly homogenous chromatographycally. Molecular weight of PPAase, as shown by means of gel filtration, was 66000, K(m) was calculated to be 1.64-10-4 M. PPAase was inhibited by SH-reagents and partially by o-oxyquinoline. Some increase in the enzyme activity was observed in the presence of Ca2+, Mg2+ and Mn2+ in low concentrations. The enzyme hydrolysed amino acid arylamides with hydrophobic side groups and some dipeptides, which had at least one hydrophobic amino acid and did not contain amino acids with polar group.
经检测，巢菜种子提取物对L-苯丙氨酰-对硝基苯胺（PPA）和甘氨酰-对硝基苯胺的水解作用是由两种不同的芳基酰胺酶引起的。其中一种PPA酶通过羟基磷灰石、DEAE-纤维素和Sephadex G-100凝胶过滤进行了2000倍纯化。得到的制剂在色谱上几乎是纯的。通过凝胶过滤显示，PPA酶的分子量为66000，计算得出的米氏常数（K(m)）为1.64×10⁻⁴M。PPA酶受到巯基试剂的抑制，并部分受到邻羟基喹啉的抑制。在低浓度的Ca²⁺、Mg²⁺和Mn²⁺存在下，观察到酶活性有所增加。该酶能水解具有疏水侧基的氨基酸芳基酰胺和一些二肽，这些二肽至少含有一个疏水氨基酸且不含带有极性基团的氨基酸。
