Structure and functional characterization of recombinant chondroitin sulphate AC lyase of family 8 polysaccharide lyase (PsPL8A) from Pedobacter saltans for its in vitro applications in therapeutics and production of chondroitin oligosaccharide with anti-cancer and prebiotic properties

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Date
2017
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Abstract
Chondroitin sulphate (CS) is a part of connective tissues and involved in major biological processes such as cell migration, proliferation, adhesion, pathogenesis, cell-matrix interactions, chemokine and cytokine activation. The CS degrading enzymes, chondroitin lyases are therapeutically important enzymes. Chondroitin AC lyase (PsPL8A) from Pedobacter saltans DSM12145 was cloned expressed functionally and structurally characterized. The time dependent TLC analysis of PsPL8A degraded products of C4S revealed that it follows initially a concomitant endo-and exo-lytic mode which finally shifted to exolytic mode of catalysis. Small angle X-ray scattering (SAXS) analysis revealed that the solution shape of PsPL8A is in good correlation with modeled structure and its homologous crystal structures. Chondroitin AC lyase (PsPL8A) showed anti-proliferative effect on human melanoma (SK-Mel 28) and fibrosarcoma (HT-1080) cell lines. PsPL8A disrupts the mitochondrial cell potential and induce apoptosis in fibrosarcoma and melanoma cells causing cell death and hence prospects as a potential agent for therapeutic approach in cancer treatment. Chondroitin sulphate was isolated from chicken keel bone and was explored as cheaper and sustainable source for isolation of CS for its future use in pharmaceutical industry. CS was physico-chemically characterised by ESI-MS, NMR, HPSEC, DLS, FESEM, AFM, TGA, DSC and DTG analyses.
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Supervisor: Arun Goyal
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BIOSCIENCES AND BIOENGINEERING
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