Stano J, Siekel P, Micieta K, Blanárikova V, Korenová M, Bergerová E, Nemec P
Garden of Medicinal Plants, Comenius University, Bratislava, Slovak Republic.
Pharmazie. 2008 Dec;63(12):909-12.
A simple, rapid and straightforward procedure for identification and determination of intracellular and extracellular activity of aminopeptidases employing synthetic substrates beta-naphtylamides of L-Ala, L-Phe, and L-Tyr was used. Poppy cells (Papaver somniferum L.) permeabilized by Tween 80 were immobilized via crosslinking by glutaraldehyde. Glutaraldehyde immobilized poppy cells lost their viability and demonstrated significantly lower aminopeptidase activities than untreated control cells probably due to a damage to the enzyme active centre. Poppy cells immobilized by pectate and alginate have retained high activity of studied aminopeptidases. The culture medium (without cells) used for the identification and determination of extracellular enzyme activities retained 20-21%, whereas intracellular activities were estimated to be 79-80% of total enzyme activity. Thus the intracellular specific activity was 1.00-1.07 higher.
采用L-丙氨酸、L-苯丙氨酸和L-酪氨酸的合成底物β-萘酰胺，使用一种简单、快速且直接的程序来鉴定和测定氨肽酶的细胞内和细胞外活性。经吐温80透化的罂粟细胞（罂粟）通过戊二醛交联固定。戊二醛固定的罂粟细胞失去了活力，与未处理的对照细胞相比，氨肽酶活性显著降低，这可能是由于酶活性中心受损。通过果胶酸盐和藻酸盐固定的罂粟细胞保留了所研究氨肽酶的高活性。用于鉴定和测定细胞外酶活性的培养基（无细胞）保留了20 - 21%的总酶活性，而细胞内活性估计占总酶活性的79 - 80%。因此，细胞内比活性高1.00 - 1.07倍。
