A non-fluorescent pigment produced from the fluorescent Bacillus endophyticus AVP-9 which possess abundant lutein content considered as a xanthophyl-type compound with potentially beneficial pharmacological properties for enhancing human health. In this context, bacterial lutein was extracted by employing different solvents to isolate from Bacillus endophyticus was examined in this study. Chromatographically purified lutein was spectrally characterized using ultraviolet-visible spectrometry and spectral analysis was carried through liquid chromatography-mass spectrometry, functional group identification was done by using fourier transform-infrared spectroscopy, structural determination by nuclear magnetic resonance spectrometry. By using hexane and ethyl acetate (7:3) with the flow rate of 5ml/min, was proven to be an excellent solvent phase for separation of lutein, meaning that up to 90% pure lutein generated with a 60% estimated yield. The purified compound identified as a β, ε-3, 31-diol (lutein) having molecular weight of 568.6g/mol with the molecular formula C40H56O2 which exhibits potential antibacterial activity against E. coli (35mm) followed by S. typhi (33mm), S. flexneri (30mm), P. aureginosa (29mm), M. lutea (28mm), P. vulgaris (24mm), S. aureus (21mm) and lowest antibacterial activity against V. cholera (19mm). Purified pigment shows concentration dependent antioxidant activities with IC50 values of pigment was found to be 4.5 mg/ml. Whereas anti-proliferation activity pigment shows considerable effect MCF-7 and LNCaP cell lines, when treated with 100µg/ml, cell viability was decreased to 36.363% at IC50 value of 21.935 µg/ml and for LNCaP, the cell viability was decreased to 42.449% at IC50 value of 56.937 µg/ml.
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