Proteolytic activity in latices of Asteraceae and Campanulaceae

Abstract

In the order Asterales only two species are known to have proteolytic activity in their latices, first Taraxalisin from dandelion Taraxacum officinale Webb s.l., and second Parthenain from Guayule Parthenium argentatum L.. Both are characterized as serine endopeptidases [1]. Proteolytic enzymes isolated from plant latex have received special attention in the pharmaceutical industry and biotechnology due to their property of being active over wide range of temperature and pH. Nearly half of the commercially available enzymes are proteases, frequently used in food processing, tenderization of meat, brewing, cheese elaboration, bread manufacturing, leather and textile industries [1]. In this investigation the latex of 40 species of the Asteraceae family and 8 species of the Campanulaceae, which are not biochemical characterized before, were collected in the Botanical Garden Berlin. To determine proteolytic activity we used the fluorogenic substrate BODIPY FL- casein (Moleculare Probes, Inc., USA) [2]. To investigate the type of endopeptidases, the latex samples were pre-incubated with specific inhibitors for serine proteases (AEBSF (4-(2-Aminoethyl)-benzenesulfonyl fluoride hydrochloride), cysteine proteases (E64 (4-(2-Aminoethyl) benzenesulfonylfluoride hydrochloride)), aspartatic proteases (Pepstatin A) and metalloprotease (EDTA) and the remaining activity was determined. In both families highly active serine proteases were found.