Characterization of Lipoxygenase in AOT/Isooctane Reversed Micelles

AOT/isooctane 역미셀계내에서의 lipoxygenase 반응 특성

  • Published : 2002.04.01

Abstract

An investigation was carried out to study the characteristics of lipoxygenase in dioctyl sulfosuccinate (aerosol-OT, AOT)/isooctane revered micelles of microaqueous system containing infinitesimal water. ${\alpha}-Linoleic$ acid as a substrate could be analyzed by the colorimetric methodology using 5%(w/v) cupric acetate-pyridine solution and the activity of lipoxygenase was able to be assayed by the degree of ${\alpha}-linoleic$ acid consumption per minute. Optimal pH, temperature, and R-value ([water]/[AOT]) were determined as the value of 5.0, $25^{\circ}C$, and 10.0, respectively. Kinetic analysis of the enzyme reaction under the optimal conditions showed that the values of $K_m$ and $V_{max}$ were 0.31 mM of ${\alpha}-linoleic$ acid and $384.16{\mu}mol$ of ${\alpha}-linoleic$ acid decomposed/min, respectively. The results indicate the reaction to be lipoxygenase-catalyzed oxidation of ${\alpha}-linoleic$ acid in AOT/isooctane reversed micellar system. The inhibitory effect of natural antioxidants on lipoxygenase showed little inhibitory effect of L-ascrobic acid while ${\alpha}-tocopherol$ showed 72% of inhibitory effect.

Keywords

lipoxygenase;reversed micelles;kinetic analysis;inhibition

References

  1. Rho, Y.T., Kim, Y.S., Cho, H.Y. and Pyun, Y.R. A study on pro-tein separation with AOT-isooctane reverse micellar system.Korean J. Food Sci. Technol. 21: 542-547 (1989)
  2. Han, D. and Rhee, J.S. Characteristics of lipase-catalyzed hydrol-ysis of olive oil in AOT-isooctane reversed micelles. Biotechnol. Bioeng. 28: 1250-1255 (1986) https://doi.org/10.1002/bit.260280817
  3. Eskin, N.A. Grossman, S. and Pinsky, A. Biochemistry of lipoxy-genase in relation to food quality. CRC Crit. Rev. Food Sci. Nutr.9: 1-40 (1977)
  4. Chang, M.J., Cho, I.Y. and Lee, S.K. Characteristics of hydroper-oxide lyase and lipoxygenase activity in cucumber (Cucumis Sati-vus) fruit. Korean J. Food Sci. Technol. 29: 314-319 (1997)
  5. Choi, S.W. and Osawa, T. Isolation and idendification of lipoxy-genase inhibitor from nutmeg spice. Food Sci. Biotechnol. 5:156-160(1996)
  6. Galliard, T. and Chan, H.W. A comprehensive treatise, In: The Biochemistry of Plants, Vol. 4, p. 131. Stumpf, P.K. (ed.), Aca-demic Press, New York, USA (1980)
  7. Choi, S.W., Kang, W.W., Chung, S.K. and Cheon, S.H. Antioxi-dative activity of flavonoids in persimmon leaves. Food Sci. Bio-technol.5:119-123(1996)
  8. Seoud, A.A.E. Acidities and basicities in reversed micellar sys-tems. In: Reverse Micelles, p. 81. Luisi, P.L. and Straub, B.E.(eds.), Plenum Press, New York, USA (1984)
  9. Boyes, S., Perera, C. and Young, H. Kiwifruit lipoxygenase:Preparation and characteristcs. J. Food Sci. 57: 1390-1395 (1992) https://doi.org/10.1111/j.1365-2621.1992.tb06866.x
  10. Fletcher, P.D.L, Freedman, R.B., Robinson, B.H., Rees, G.D., and Schmacker, R. Lipase-catalyzed ester synthesis in oil-continuous microemulsions. Biochim. Biophys. Acta 912: 278-282 (1987) https://doi.org/10.1016/0167-4838(87)90098-7
  11. Eicke, H.F. and Shepherd, J.C.W. Dielectric properties of apolar micelle solutions containing solubilized water. Helv. Chim. Acta 57: 1951-1963 (1974) https://doi.org/10.1002/hlca.19740570708
  12. Im, H.S., Cho, Y.H. and Rhee, C.O. Effect of inhibitor on lipoxy-genase inactivation in soybean homogenates. Korean J. Food Sci. Technol. 27: 19-24 (1995)
  13. Zakut, R. Grossman, S. Pinsky, A. and Wilchek, M. Evidence for an essential methionine residue in lipoxygenase. FEBS Lett. 72:107-110(1976)
  14. Chang, P.S., Kang, S.T. and Rhee, J.S. A general kinetic model for lipase-catalyzed glycerolysis of triolein. Food Sci. Biotechnol 1:46-49(1992)
  15. Zaks, A. and Klibanov, A.M. Enzymatic catalysis in organic media at $100^\circ C$. Science 224: 1249-1251 (1984) https://doi.org/10.1126/science.6729453
  16. Nakayama, T., Takeura, Y. and Ueda, U. Visible spectrophotomet-ric assay, Purification and molecular properties of a lipoxygenase from egg plant (Solanum melongena Linne) fruits. Biochem. Bio-phys. Res. Commun. 214: 1067-1072 (1995) https://doi.org/10.1006/bbrc.1995.2394
  17. Kim, D.K., Han, K.Y. and Noh, B.S. Characteristics of crude lipoxygenase in Chinese cabbages. Korean J. Food Sci. Technol.29: 710-715 (1997)
  18. Shipe, W.F, Senyk, G.F. and Fountain, K.B. Modification of cupric sulfate method for the analysis of free fatty acids, J. Dairy Sci. 63: 193-198 (1980) https://doi.org/10.3168/jds.S0022-0302(80)82913-4
  19. Eicke, H. F. On the stabilization of micelles in apolar solvents by solubilized ions and polar liquids. J. Coll. Interface Sci. 52: 65-76 (1975) https://doi.org/10.1016/0021-9797(75)90303-3
  20. Lee, Y.C. Lipoxygenase and off-flavor development in some fro-zen foods. Korean J. Food Sci. Technol. 17: 53-58 (1981)
  21. Grossman, S., Trop, M., Avtalion, R. and Pinsky, A. Properties of agarose-bound soya-bean lipoxygenase. Biochem. J. 127: 909-910(1972) https://doi.org/10.1042/bj1270909
  22. Barbaric, S. and Luisi, P.L. Micellar solubilization of biopolymers in organic solvents. 5. Activity and conformation of $\alpha$-chymot-rypsin in isooctane-AOT reversed micelles. J. Am. Chem. Soc. 103:4239-4244(1981) https://doi.org/10.1021/ja00404a044
  23. Lowry, R.R. and Tinsley, I.J. Colorimetric analysis of free fatty acids by cupric sulfate, J. Am. Oil Chem. Soc. 53: 470-472(1976) https://doi.org/10.1007/BF02636814
  24. Kruger, J.E., Lineback, D. and Stauffen, C.E. Enzymes and Their Role in Cereal Technology. American Association of Cereal Chemists, Inc., p. 241 (1987)
  25. Chang, P.S. Hydrolysis of phosphatidylcholine in aerosol-OT/isooctane reversed micelles by phospholipase $A_2$. Korean J. Food Sci. Technol. 29: 26-31 (1997)
  26. Chang, P.S., Rhee, J.S. and Kim, J.J. Continuous glycerolysis of olive oil by immobilization on liposome in reversed micelles. Biotechnol. Bioeng. 38: 1159-1165 (1991) https://doi.org/10.1002/bit.260381007
  27. Grandi, C., Smith, R.E., and Luisi, P.L. Micellar solubilization of biopolymers in organic solvents: Activity and conformation of lysozyme in isooctane reverse micelles. J. Biol. Chem. 256: 837-843(1981)
  28. Grosch, W. and Laskawy, G. Differences in the amount and range of volatile carbonyl compounds fonned by lipoxygenase isoen-zymes from soybeans. J. Agric. Food Chem. 23: 791-794 (1975) https://doi.org/10.1021/jf60200a003
  29. Shin, H.S. The Newest Food Chemistry, p. 445. Kwangmungak,Seoul(1995)