Lipoxygenase of Thermoactinomyces vulgaris,purification and characterization of reaction products |
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| Institution: | 1. Institute of Biochemistry, University Medicine Berlin — Charite, Chariteplatz 1, CCO-Building, Virchowweg 6, D-10117 Berlin, Germany;2. Neuroprotection Research Laboratory, Department of Radiology, Massachusetts Genrel Hospital and Harvard Medical School, Charlestown, MA, USA |
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| Abstract: | - 1.1. A lipoxygenase preparation was obtained from Thermoactinomyces vulgaris and was purified by affinity chromatography on a linoleyl aminoethyl sepharose column.
- 2.2. Two active fractions were obtained.
- 3.3. The fraction obtained by elution with 100 mM borate buffer pH 9.0 was used in the subsequent work.
- 4.4. Th. vulgaris lipoxygenase oxidized linoleic acid into two products: 13-HPOD and 9-HPOD at a ratio of 44 to 56, respectively.
- 5.5. The identification and characterization of the isomers was done by HPLC, I.R. and mass spectrometry.
- 6.6. When arachidonic acid was used as substrate, 15-HPETE and 15-HETE were found to be the main enzymatic products.
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