Biochemical Characteristics and Healing Activity of Bromelia laciniosa Leaf Protease

Abstract

Macambira plant (Bromelia laciniosa Mart. Ex Shult. & Schult. f.) is a species belonging to the family Bromeliaceae, which is distributed widely in the hinterlands of northeastern Brazil, from Bahia to Piauí states. The plants of this species contain an enzyme complex named bromelain, which has great potential for therapeutic applications. In the present study, bromelain derived from Bromelia laciniosa leaves was assessed for its thermodynamic properties and in vitro cytotoxic and healing activities. Bromelain was extracted from the leaves and partitioned using ethanol. The partially purified bromelain was then characterized to determine the optimum pH and temperature and the kinetic and thermodynamic parameters. The partially purified bromelain derived from macambira leaves exhibited its highest activity at 47°C and pH 7, when the Km and Vmax values of 2.26 µM and 2.083 U/mL, respectively. The half-life of this bromelain at 47°C was 5.78 h. At 47°C, the Gibbs's free energy, enthalpy, and entropy values reached 89, 53.54, and –0.11 kJ/mol K, respectively. In the SDS-PAGE analysis, bromelain appeared as the main protein band, with an estimated molecular mass of 33 kDa. The in vitro cytotoxic activity and wound healing effects were determined using the MTT assay and Scratch assay, respectively. These assays were performed on the non-neoplastic cell lines L-929 and PBMC and the human tumor cell lines MCF-7, HCT-116, and NCI-H292. Bromelain exhibited a positive healing activity with a change rate similar to that observed for the control. In addition, bromelain exhibited cytotoxic activity against NCI-H292 tumor cells and chemopreventive potential. These results indicate that bromelain derived from macambira leaves may be utilized as a potential ingredient in pharmaceutical formulations, for various healing and chemopreventive purposes and as an alternative to the bromelain extracted from pineapple. In addition, the biochemical characteristics of this bromelain could find application in the food industry. Future studies should, therefore, focus on deciphering the action mechanisms underlying the activities of bromelain observed in the present study to further confirm and enhance its application potential for therapeutic purposes.