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. 1992 Dec;100(4):1802–1807. doi: 10.1104/pp.100.4.1802

Purification and Partial Characterization of a Membrane-Associated Lipoxygenase in Tomato Fruit 1

Caroline G Bowsher 1,2,3,2, Bonita J M Ferrie 1,2,3, Sibdas Ghosh 1,2,3, James Todd 1,2,3,3, John E Thompson 1,2,3, Steven J Rothstein 1,2,3
PMCID: PMC1075867  PMID: 16653200

Abstract

Membrane-associated lipoxygenase from green tomato (Lycopersicon esculentum L. cv Caruso) fruit has been purified 49-fold to a specific activity of 8.3 μmol·min−1·mg−1 of protein by solubilization of microsomal membranes with Triton X-100, followed by anion- exchange and size-exclusion chromatography. The apparent molecular mass of the enzyme was estimated to be 97 and 102 kD by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and size-exclusion chromatography, respectively. The purified membrane lipoxygenase preparation consisted of a single major band following sodium dodecyl sulfate-polyacrylamide gel electrophoresis, which cross-reacts with immunoserum raised against soluble soybean lipoxygenase 1. It has a pH optimum of 6.5, an apparent Km of 6.2 μm, and Vmax of 103. μmol·min−1·mg−1 of protein with linoleic acid as substrate. Corresponding values for the partially purified soluble lipoxygenase from tomato are 3.8 μm and 1.3 μmol·min−1·mg−1 of protein, respectively. Thus, the membrane-associated enzyme is kinetically distinguishable from its soluble counterpart. Sucrose density gradient fractionation of the isolated membranes indicated that the membrane-associated lipoxygenase sediments with thylakoids. A lipoxygenase band with a corresponding apparent mol wt of 97,000 was identified immunologically in sodium dodecyl sulfate-polyacrylamide gel electrophoresis-resolved proteins of purified thylakoids prepared from intact chloroplasts isolated from tomato leaves and fruit.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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