Kinetic characterization of a semipurified preparation from bromelain for antitumor use

[摘要] INTRODUCTION: most of the world production of enzymes is devoted to proteases obtaining. Applications of these biomolecules for therapeutical objectives are very varied. Some are known as natural remedies and other are drugs modifying the biological response. Pineapple plants contain some cysteine-proteases, the majority component isolated from stem is the "stem Bromelain" (EC 3.4.22.32). In the health field it has attributed some actions including the differentiation induction of tumor cells and attenuation of tumor growth. OBJECTIVES: to characterize kinetically a semipurified preparation from Bromelain obtained by a procedure developed in Cuba to be assessed in the health field. METHODS: the crude extract from Bromelain obtained from the adult plant stems of Ananas comosus (L.) Merr. Cv. Spanish red (pineapple) was purified by ion-exchange chromatography in carboxylmethylcelulose-52 in semibatch system. RESULTS: kinetic characterization of purified extract allowed us to verify the existence of a stable and active preparation with a specific activity of 0,94 U/mg of proteins (2,34 times superior to that of original crude extract) with a protein fraction in the main of molar mass 24 500 Da and a isoelectric point, located in the basic pH zone (9,55). It has a 6,8optimal pH using the 2% hemoglobin as substrate at 37ºC and stability depending on pH and on acceptable temperature. If it is preserved lyophilized at 20 ºC it maintains the 80% of activity for a year. CONCLUSIONS: the ion-exchange chromatography in carboxyl methylcellulose-52 allowed the achievement of semipurified preparation from Bromelain with a specific activity higher to that of original extract that may to be used in medicine as antitumor agent.

[学科分类] 药理学

[关键词] cisteíno-proteasas;bromelina;Ananas comosus