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Isolation of Indole-3-ethanol Oxidase from Cucumber Seedlings

Larry E. Vickery, William K. Purves
Larry E. Vickery
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William K. Purves
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Published May 1972. DOI: https://doi.org/10.1104/pp.49.5.716

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Abstract

Previous work in this laboratory has shown that cucumber (Cucumis sativus L.) seedlings contain large amounts, relative to other indolic compounds, of extractable indole-3-ethanol (IEt); tracer studies have established that IEt is metabolized to IAA. We have now succeeded in isolating an enzyme from these seedlings which catalyzes the oxidation of IEt to indole-3-acetaldehyde (IAAld). The identification of the product as IAAld was based on solvent partitioning of the free aldehyde and its bisulfite adduct and radiochromatography following incubation of enzyme with 14C-IEt. A novel, quantitative colorimetric test for IAAld was also developed utilizing the Salkowski reagent. Partial purification of the enzyme was achieved by salt gradient chromatography on Bio-Rex 70, heating the preparation to 70 C, and chromatography on Sephadex G-150. This purification procedure yielded an enzyme activity purified in excess of 3000-fold, and studies on a standardized Sephadex column suggest a molecular weight of the enzyme of approximately 105,000. The reaction was found to proceed only aerobically; and, in the absence of other electron acceptors, O2 appears to be reduced to H2O2. The enzyme has nearly maximum activity from pH 8 to 11.

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Isolation of Indole-3-ethanol Oxidase from Cucumber Seedlings
Larry E. Vickery, William K. Purves
Plant Physiology May 1972, 49 (5) 716-721; DOI: 10.1104/pp.49.5.716

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Isolation of Indole-3-ethanol Oxidase from Cucumber Seedlings
Larry E. Vickery, William K. Purves
Plant Physiology May 1972, 49 (5) 716-721; DOI: 10.1104/pp.49.5.716
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Plant Physiology
Vol. 49, Issue 5
May 1972
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