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Preparation of (-)-5,6,7,8-tetrahydrofolate using immobilized dihydrofolate reductase.

作者信息

Ahmed F, Dunlap R B

出版信息

Anal Biochem. 1984 Aug 15;141(1):149-54. doi: 10.1016/0003-2697(84)90438-x.

DOI:10.1016/0003-2697(84)90438-x
PMID:6496924
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.

摘要

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