Purification and Characterization of an Endoprotease from Melon Fruit

An endoprotease was purified from melon fruit (Cucumis melo L.) by ammonium sulfate precipitation, gel filtration and ion-exchange chromatography using t-butyloxycarbonyl-Ala-Ala-Pro-Leu p-nitroanilide as a substrate. The molecular weight was estimated as 26,000 and isoelectric point pH 9.5. It preferentially hydrolyzed peptide bonds of the carboxyl terminal sides of Leu, Ala, His, Gin, and Am. Activity was strongly inhibited by diisopropyl phosphofluoridate, indicating the serine protease nature of the enzyme. The migration distance on electrophoresis, molecular weight and substrate specificity differed from cucumisin, a known protease from melon. This unusual protease may have potential for special food treatment applications.