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Preparation of levo 5 6 7 8 tetrahydrofolate using immobilized dihydrofolate reductase ec

, : Preparation of levo 5 6 7 8 tetrahydrofolate using immobilized dihydrofolate reductase ec Analytical Biochemistry 141(1): 149-154

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 UV and circular dichroic spectra and its reactivity as a cofactor in the thymidylate synthetase reaction.

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