Abstract
Pectin was extracted from Citrus reticulata (mandarin) peels by ultrasound assisted extraction process. The yield of extracted mandarin pectin (EMP) was 21.3 ± 1.2%. EMP after purification (pEMP) by size-exclusion chromatography gave molecular mass, 92.3 kDa. Field-emission scanning electron microscopy image of pEMP showed wrinkled surface and Atomic force microscopy revealed fibrous network. Fourier transform infrared spectroscopic and nuclear magnetic resonance analyses of pEMP showed hetero-polysaccharide nature with 68 ± 2% methylated galacturonic acids. Thermogravimetric analysis showed degradation at 225 ± 5 °C. pEMP treated by recombinant clostridial pectate-lyase (CtPL1B) or a mixture of CtPL1B and pectin-methylesterase (CtPME) produced pectic-oligosaccharides having degree of polymerization, DP2 and DP3. Enzyme assays displayed that CtPME hydrolyzed methyl groups from pEMP and resulting pectate was degraded by CtPL1B. ESI-MS analysis confirmed that the treatment of pEMP by CtPL1B produces methylated unsaturated di-galacturonic and tri-galacturonic acids, which, displayed biocompatibility with normal human kidney (HEK293) cells and cytotoxicity against colon cancer (HT29) cells.
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