Purification and identification of bioactive protein from leaves of Datura inoxia P.mil

Abstract

A protein with bioactive potential was purified from the leaves of Datura inoxia P.mil. The preliminary screening of the crude proteins fractionated by Ammonium Sulphate was carried out for its antibacterial, antioxidant and anticancer properties. It exerted antibacterial activity against three bacteria namely Vibrio cholerae, Staphylococcus aureus, and Escherichia coli but was not effective against Aeromonas hydrophilia. It unveiled an increasing free radical scavenging activity in a dose-dependent manner and presented a potential anticancer activity against human colon adenocarcinoma cell line HT 29. The active protein fraction was purified by ion exchange chromatography on CM-Sephadex. The purified protein has a molecular weight of 14.3 kDa and Peptide Mass Fingerprint using Matrix-Assisted Laser Desorption/Ionization-Time Of Flight Mass Spectroscopy (MALDI-TOF/MS) revealed peptide matches to defensin-like protein of Brassica napus, chaperonin CPN60-2 and mitochondria of Cucurbita maxima. The purified protein showed anticancer activity against HT 29 cancer cell line.