摘要
A competitive inhibitor (K i = 10 -4 -10 -5 m) for lactic acid dehydrogenase (LDH), N-(6-aminohexyl)oxamate, was synthesized from diethyl oxalate and 1,6-diaminohexane. The affinity column prepared by attachment of this compound to CNBr-activated Sepharose 4B enabled efficient purification of LDH from the marine teleost, Fundulus heteroclitus (Lin.). A similar affinity absorbent prepared by the method of O'Carra and Barry (FEBS Lett. (1972) 21, 281) gave a less satisfactory purification because the LDH was contaminated with phosphorylase a. The reason for this contamination was traced to an incomplete acylation of amino groups on the modified Sepharose beads by the carbodiimide-mediated reaction. Immobilization of the presynthesized affinant, N-(6-aminohexyl)oxamate, assured complete absence of these interfering groups. An alternate synthetic scheme, involving an active ester of ethyl oxalate, was also found to eliminate this problem. © 1977.