凤梨蛋白酶活性成分的再研究。

Reinvestigation of the proteolytically active components of Bromelia pinguin fruit.

作者信息

Payrol Juan Abreu, Obregón Walter D, Natalucci Claudia L, Caffini Néstor O

机构信息

Instituto de Farmacia y Alimentos, Universidad de La Habana,Calle 23, Entre 214 y 222, # 21425, La Coronela, La Lisa, Ciudad de La Habana, CP 13600, Cuba.

出版信息

Fitoterapia. 2005 Sep;76(6):540-8. doi: 10.1016/j.fitote.2005.04.021.

DOI:10.1016/j.fitote.2005.04.021

PMID:15978746

Abstract

Pinguinain is the name given to a proteolytic enzyme preparation obtained from Bromelia pinguin fruits that has been scarcely studied. The present paper deals on the reexamination of the proteases present in fruits of B. pinguin grown in Cienfuegos, Cuba. The preparation (partially purified pinguinain, PPP) showed the main characteristics of the cysteine proteases, i.e., optimum pH within alkaline range (pH 7.2-8.8), inhibition of proteolytic activity by thiol blocking reagents, which is usually reverted by addition of cysteine, a remarkable thermal stability and notable stability at high ionic strength values. Isoelectric focusing and zymogram of PPP revealed the presence of several proteolytic components between pI 4.6 and 8.1. Preliminary peptidase purification by cationic exchange chromatography showed the presence of two main proteolytic fractions with molecular masses of approximately 20.0 kDa, according to SDS-PAGE.

摘要

凤梨蛋白酶是从几乎未被研究过的凤梨科植物果实中获得的一种蛋白水解酶制剂的名称。本文对古巴西恩富戈斯种植的凤梨科植物果实中存在的蛋白酶进行了重新研究。该制剂（部分纯化的凤梨蛋白酶，PPP）显示出半胱氨酸蛋白酶的主要特征，即碱性范围内的最佳pH值（pH 7.2 - 8.8）、硫醇封闭试剂对蛋白水解活性的抑制作用，通常通过添加半胱氨酸来恢复，具有显著的热稳定性和在高离子强度值下的显著稳定性。PPP的等电聚焦和酶谱显示在pI 4.6和8.1之间存在几种蛋白水解成分。根据SDS - PAGE，通过阳离子交换色谱法进行初步肽酶纯化显示存在两个主要的蛋白水解级分，分子量约为20.0 kDa。