Purification and Identification of Squalene Synthase Inhibitor Isolated from Fermented Soybean Paste

  • Published : 2005.02.28


Squalene synthase (SQS) inhibitors were screened from various plants and food extracts. Effective SQS inhibitor was purified from fermented soybean paste using ethanol extraction, HP-20 column chromatography, ethyl acetate extraction, silica gel column chromatography, and crystallization. Through UV spectrometry, $^1H$ NMR, $^{13}C$ NMR, and mass spectrometry, SQS inhibitor was identified as daidzein with molecular mass of 254 and molecular formula of $C_{15}H_{10}O_4$. Daidzein showed $IC_{50}$ value of 50 nmol/L against SQS, confirming its potential as therapeutic agent for hypercholesterolemia.


  1. J. Med. Chem. v.34 Isoprenyl phosphinylformates: new inhibitors of squalene synthetase Biller, S.A.;Forster, C.;Gordon, E.M.;Harrity, T.;Rich, L.C.;Marretta, J.;Ciosek, C.P. Jr. https://doi.org/10.1021/jm00110a024
  2. J. Lipid Res. v.26 Membrane lipid composition and cellular function Spector, A.A.;Yorek, M.A.
  3. Bioorg. Med. Chem. v.11 Syntheses of 3-ethylidenequinuclidine derivatives as a squalene synthase inhibitors. Part 2: enzyme inhibition and effects on plasma lipid levels Ishihara, T.;Kakuta, H.;Moritani, H.;Ugawa, T.;Sakamoto, S.;Tsukamoto, S.;Yanagisawa, I. https://doi.org/10.1016/S0968-0896(03)00336-5
  4. Biochim. Biophy. Acta. v.1170 Regulation of isoprenoid metaboliosm in rat liver: near constant chain lengths of dolichy phosphate and ubiquinone are maintained during greatly altered rates of cholesterogenesis Keller, R.K.;Vilsaint, F.
  5. Eur. J. Pharmacol. v.431 Squalene synthase inhibitors reduce plasma triglyceride through a lowdensity lipoprotein receptor-independent mechanism Hiyoshi, H.;Yanagimachi, M.;Ito, M.;Saeki, T.;Yoshida, I.;Okada, T.;Ikuta, H.;Shinmyo, D.;Tanaka, K.;Kurusu, N.;Tanaka, H. https://doi.org/10.1016/S0014-2999(01)01450-9
  6. Biosynthesis of Isoprenoid Compounds Poulter, C.D.;Rilling, H.C.
  7. Proc. Natl. Acad. Sci. v.90 Zaragozic acids: A family of metabolites that are picomolar competitive inhibitors of squalene synthase Bergstrom, J.D.;Krutz, M.M.;Rew, D.J.;Amend, A.M.;Karkas, J.D.;Bostedor, R.D.;Bansal, V.S.;Dufresns, C.;Middleworth, F.L.Y.;Hensens, O.D.;Liesch, J.M.;Zinc, D.L.;Wilson, K.E.;Onishi, J.;Milligan, J.A.;Bills, G.;Kaplan, L.;Nallin, M.O.;Jenskins, R.G.;Huang, L.;Meinz, M.S.;Quinn, L.;Burg, R.U.;Kong, Y.L.;Mochales, S.;Mojena, M.;Martin, I.;Pelaez, F.;Diez, M.T.;Albert, A.W.
  8. Methods Enzymol. v.110 Squalene synthetase Agnew, W.S. https://doi.org/10.1016/S0076-6879(85)10094-7
  9. J. BioI. Chem. v.262 Squalene synthetase. Solubilization and partial purification of squalene synthetase, copurification of presqualene pyrophosphate and squalene synthetase activities Kuswik, R.G.;Rilling, H.C.
  10. Biochem. Biophy. Res. Commun. v.138 A Regulation of squalene synthase in rat liver Cohen, L.H.;Griffioen, A.M.;Wanders, R.J. https://doi.org/10.1016/0006-291X(86)90285-8
  11. Arch. Biochem. Biophy. v.152 Hog liver squalene synthase: The partial purification of the particulate enzyme and kinetic analysis of the reaction Dugan, R.E.;Porter, J.W. https://doi.org/10.1016/0003-9861(72)90189-0
  12. J. Antibiotics v.45 The squalestatins, novel inhibitors of squalene synthase produced by a species of Phoma. I. Taxonomy, fermentation, isolation, physico-chemical properties and biological activity Dawson, M.J.;Farthing, J.E.;Marshall, P.S.;Middleton, R.F.;O'Neill, M.J.;Shuttleworth, A.;Stylli, C.;Tait, R.M.;Taylor, P.M.;Wildman, H.G.;Buss, A.D.;Langley, D.;Hayes, M.Y. https://doi.org/10.7164/antibiotics.45.639
  13. Food Chem. Toxic. v.26 Reduction of benzo(a)pyren-induced mouse forestomach neoplasms in mice given nitrite and dietary soy sause Benjamin, H.;Storkson, J.;Tallas, P.G.;Pariza, M.W. https://doi.org/10.1016/0278-6915(88)90066-X
  14. J. Kor. Soc. Food Nutr. v.22 Antioxidant characteristics of melanidin related products fractionated from fermented soybean sauce Cheigh, H.S.;Lee, J.S.;Moon, G.S.;Park, K.Y.
  15. J. Agric. Food Chem. v.41 Genistein, daidzein, and their ${\beta}-glycoside$ conjugates: antitumor iso-flavones in soybean foods from American and Asian diets Coward, L.;Barnes, N.C.;Stechell, K.D.R.;Barnes, S. https://doi.org/10.1021/jf00035a027
  16. Cancer Res. v.43 Bowman-Birk soybean protease inhibitor as an anticarcinogen Yavelow, J.;Finalay, T.H.;Kennedy, A.R.;Troll, W.
  17. Carcinogenesis v.10 Inositol and inositol hexaphosphate supree cell proliferation and tumor formation in CD-1 mice Shamsuddin, A.M.;Ullah, A.;Chakravarthy, A.K. https://doi.org/10.1093/carcin/10.8.1461
  18. Food Sci. Biotechnol. v.11 no.4 Characteristics of taste components of commercial soybean paste Park, H.K.;Gil, B.I.;Kim, J.K.
  19. J. BioI. Chem. v.267 Squalestatin I, a potent inhibitor of squalene synthase, which lower serum cholesterol in vivo Baxter, A.;Fitzgerald, B.J.;Hutson, J.L.;McCarthy, A.D.;Motteram, J.M.;Ross, B.C.;Sapra, M.;Snowden, M.A.;Watson, N.S.;Williarm, R.J.;Wright, C.