Preparation of high-specific-activity D-[3-3H]pantothenic acid |
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Authors: | D S Vallari C O Rock |
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Affiliation: | 1. Department of Biochemistry, St. Jude Children''s Research Hospital, Memphis, Tennessee 38101 USA;2. The University of Tennessee, Memphis, Tennessee 38101 USA |
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Abstract: | High-specific-activity D-[3-3H]pantothenic acid (5 Ci/mmol) was prepared from commercially available beta-[3-3H]alanine employing Escherichia coli strain DV1 (panD2 pan F1). This strain is defective in beta-alanine synthesis and pantothenate uptake, and under appropriate growth conditions converted 85 to 90% of the input beta-[3-3H]alanine to extracellular D-[3-3H]pantothenate. The radiolabeled vitamin was purified from the medium by thin-layer chromatography followed by reverse-phase high-performance liquid chromatography. The overall yield of D-[3-3H]pantothenic acid was 30% and radiochemical purity was greater than 99%. |
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