Preparation of (-)-5,6,7,8-tetrahydrofolate using immobilized dihydrofolate reductase |
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Authors: | F Ahmed R B Dunlap |
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Affiliation: | Department of Chemistry, University of South Carolina, Columbia, South Carolina 29208 USA |
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Abstract: | Dihydrofolate reductase from methotrexate-resistant Lactobacillus casei was immobilized on carbodiimide-activated CH-Sepharose. The immobilized enzyme was utilized in the synthesis of (-)-5,6,7,8-tetrahydrofolate from dihydrofolate and NADPH in a batchwise reaction system. The products of the reaction, (-)-tetrahydrofolate and NADP+, were separated on a Sephadex G-10 column equilibrated with 50 mM NH4HCO3 containing beta-mercaptoethanol and ethanol. The tetrahydrofolate was then characterized by ultraviolet and circular dichroic spectra and its reactivity as a cofactor in the thymidylate synthetase reaction. |
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Keywords: | tetrahydrofolate synthesis dihydrofolate reductase enzyme immobilization |
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