Purification and properties of Pinus taeda arginase from germinated seedlings |
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Authors: | Pinus taeda |
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Affiliation: | a Department of Biological Sciences, University of Alberta, Edmonton, Alberta, T6G-2E9, Canada;b Institute of Food and Agricultural Sciences, School of Forest Resources and Conservation, University of Florida, Gainesville, Florida, 32611, USA |
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Abstract: | In germinated loblolly pine (Pinus taeda L.) seeds arginine accumulates in the seedling during its growth immediately following germination. The enzyme arginase (L-arginine amidinohydrolase, EC 3.5.3.1) is responsible for hydrolyzing this arginine into ornithine and urea. Loblolly pine arginase was purified to homogeneity from seedling cotyledons by chromatographic separation on DE-52 cellulose, Matrex Green and arginine-linked Sepharose 4B. The enzyme was purified 148-fold and a single polypeptide band was identified as arginase. The molecular mass was determined to be 140 kDa by FPLC, while the subunit size was shown to be 37 kDa by SDS-PAGE, predicting a homotetramer holoprotein. Removal of manganese from the enzyme abolishes catalytic activity, which can be restored by incubating the protein with Mn2+. Antibodies, raised against the arginase subunit, are able to immunotitrate arginase activity and are monospecific for arginase on immunoblots. |
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Keywords: | arginase conifers germination protein purification Pinus taeda reserve mobilization |
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