Purification and Properties of Lipoxygenase-like Enzyme from a Red Alga, Porphyra yezoensis

Lipoxygenase-like enzyme was purified 17-fold in a yield of 15% from crude enzyme extract of a red alga Porphyra yezoensis to near homogeneity by heat treatment, stepwise elution with DE-52 cellu lose and a Sephadex G-75 gel filtration column chromatography. The final preparation showed a single band with polyacrylamide gel electrophoresis, and its specific staining for lipoxygenase activity existed on the same site. The estimated specific activity of the purified enzyme was 34.4 ƒÊmol/min per mg pro tein, using linoleic acid as a substrate. The purified enzyme had a pH optimum of pH 9 and was found to be highly resistant to heat treatment. The activity of the purified enzyme was inhibited by hydrogen peroxide, but not by cyanide and EDTA. The purified enzyme had a higher specificity for linoleic acid. It was assumed from the results that the enzyme purified from Porphyra yezoensis is closer to a lipox ygenase and not a hematin compound.

[1]  W. Gerwick,et al.  Novel oxylipins from the temperate red alga polyneura latissima: Evidence for an arachidonate 9(S)-lipoxygenase , 1997, Lipids.

[2]  S. Rosahl.  Lipoxygenases in Plants -Their Role in Development and Stress Response , 1996, Zeitschrift fur Naturforschung. C, Journal of biosciences.

[3]  Liu Qy,et al.  Isolation of a gametophyte-specific cDNA encoding a lipoxygenase from the red alga Porphyra purpurea. , 1994 .

[4]  M. Reith,et al.  Isolation of a gametophyte-specific cDNA encoding a lipoxygenase from the red alga Porphyra purpurea. , 1994, Molecular marine biology and biotechnology.

[5]  M. Hamberg,et al.  Biosynthesis of vicinal dihydroxy fatty acids in the red alga Gracilariopsis lemaneiformis: identification of a sodium-dependent 12-lipoxygenase and a hydroperoxide isomerase. , 1993, Archives of biochemistry and biophysics.

[6]  W. Gerwick,et al.  12-Lipoxygenase activity in the red marine alga Gracilariopsis lemaneiformis , 1990 .

[7]  J. Beneytout,et al.  Properties of a Lipoxygenase in Green Algae (Oscillatoria sp.). , 1989, Plant physiology.

[8]  S. Grossman,et al.  Determination of the activity of lipoxygenase (lipoxidase). , 2006, Methods of biochemical analysis.

[9]  J. Bonnet,et al.  LIPOXYGENASE FROM TOMATO FRUIT: PARTIAL PURIFICATION AND STUDY OF SOME PROPERTIES , 1977 .

[10]  Y. Matsuda,et al.  Purification and Properties of Co2+ Requiring Heme Protein Having Lipoxygenase Activity from Fusarium oxysporum , 1976 .

[11]  M. Takashio,et al.  Isolation of Lipoxygenase-like Enzyme from Fusarium oxysporum , 1976 .

[12]  B. Vick,et al.  Lipoxygenase in Chlorella pyrenoidosa. , 1973, Lipids.

[13]  S. Svensson,et al.  Denatured hemoproteins as catalysts in lipid oxidation , 1971, Journal of the American Oil Chemists' Society.

[14]  P. Budowski,et al.  Linoleate oxidation induced by lipoxygenase and heme proteins: a direct spectrophotometric assay. , 1970, Analytical biochemistry.

[15]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.