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Characterization and Immobilization of Marine Algal 11-Lipoxygenase from Ulva fasciata

  • Original Paper
  • Published:
Journal of the American Oil Chemists' Society

Abstract

The lipoxygenase (LOX) of the marine green alga Ulva fasciata was purified and immobilized in order to improve the stability and reusability. The algal LOX was partially purified by fractionation with 35–55% saturation of ammonium sulfate and MacroPrep high Q anion exchange chromatography. The LOX was purified ten times using linoleic acid (C18:2) or arachidonic acid (C20:4) as substrate, the Michaelis constant (K m) of LOX was 117.6, 31.3 μM, and maximum velocity (V max) was 12.8, 23.3 μmol hydroperoxy fatty acid/min-mg protein, respectively. The algal LOX showed the highest activity towards C18:4 followed by C20:4, C18:2 and methyl ester of C18:4. LOX activity increased up to 10.5 times with increased concentration of Triton X-100 in the extraction medium reaching an optimum at 0.05%. Calcium chloride, glutathione and phenylmethylsulphonyl fluoride were found effective protectants to LOX during purification. Hydroperoxyeicosatetraenoic acid (HpETE) formed from arachidonic acid catalyzed by this purified algal LOX was reduced and identified as 11-hydroxy-5,8,12,14-eicosatetraenoic acid (11-HETE) by NP-HPLC and GC–MS. This algal 11-LOX was immobilized in alginate beads. The stability was sevenfold greater than that of the unbound lipoxygenase at 4 °C in 0.05 M Tris–HCl buffer (pH 7.5). This is the first report on immobilization of a marine algal lipoxygenase with a view to its potential role in seafood flavor formation.

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Acknowledgments

The taxonomy of the marine algae was identified by Dr. Su-fang Huang of the Department of Botany, National Taiwan Museum. The study was funded by the Fisheries Agency, Council of Agriculture, Taipei, Taiwan, ROC.

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Authors and Affiliations

  1. Department of Food Science, National Taiwan Ocean University, 2 Pei-Ning Road, Keelung, 202, Taiwan, ROC

    Cheng Jung Tsai, Wu Feng Li & Bonnie Sun Pan

  2. Department of Food Science, Ching Kuo Institute of Management and Health, Keelung, Taiwan, ROC

    Cheng Jung Tsai

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  1. Cheng Jung Tsai
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  2. Wu Feng Li
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Correspondence to Bonnie Sun Pan.

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Tsai, C.J., Li, W.F. & Pan, B.S. Characterization and Immobilization of Marine Algal 11-Lipoxygenase from Ulva fasciata . J Am Oil Chem Soc 85, 731–737 (2008). https://doi.org/10.1007/s11746-008-1262-6

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  • DOI: https://doi.org/10.1007/s11746-008-1262-6

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