114E-15 |
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M. R. KIM1, E. J. Son1, and C. H. Lee2. (1) Department of Food Science & Nutrition, Chungnam National University, Taejon, 305-764, South Korea, (2) Korea Research Institution of Bioscience and Biotechnology, Taejon, South Korea Apoptosis, programmed cell death, is involved in a wide range of biological and pathological process such as embryogenesis, immune responses, neurodegenerative disorders, and the progression of cancer. Caspase-3, a major protease of apoptosis cascade, plays an important in the induction of apoptosis in human leukemia cell. The objective of this study is to isolate and identify inhibitory compound from natural sources. Human leukemia cell line, U937 cells cultured in RPMI 1640 was used to assay for caspase-3 inducing-inhibitory activity, which was assayed according to the degree of formation of AFC detected by spectrofluorometer. Cell viability was assayed by PMS/MTS reagent. Because F020311 among 500 species of fungi fermentation broth screened , was found to showed a high level of inhibitory activity, F020311 was fermented in YpSs liquid media for 72 hrs at 28°É in 5 L fermentor at 28°É. The inhibition compound was purified from fermentation broth and separated from hexan, ethyl acetate, butanol and water succesively. Among solvent extracts, ethyl acetate extract showed the high inhibitory activity against caspase-3, the ethyl acetate extract was further purified using silica gel column chromatography, sephadex LH20 and HPLC. The isolated compound with strong inhibitory activity was identified as penicillic acid by spectroscopic methods of ESI-MS, 1H-NMR. Penicillic acid was showed caspase-3 inducing inhibitory activity by etoposide in U937 cells in concentration dependently, and 13.56 mM of IC50 value. Cell viability assay was shown about 80% of cells viability after exposure to 100 mM of penicillic acid for 24 hrs. Natural product like F020311 fungi fermentation broth is suggested to be a useful functional source for aptosis inducing inhibitory agent.
Session 114E, Nutraceuticals & Functional Foods: Bioactivity measurement and mechanism
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