Zappelli P, Rossodivita A, Re L
Eur J Biochem. 1975 Jun;54(2):475-82. doi: 10.1111/j.1432-1033.1975.tb04159.x.
Alkylation at N-1 of the NAD+ adenine ring with 3,4-epoxybutanoic acid, followed by chemical reduction to the alkali-stable NADH form and alkaline Dimroth rearrangement, gave the NADH derivative alkylated at the exocyclic adenine amino group. Enzymic reoxidation of the latter derivative gave nicotinamide-6-(2-hydroxy-3-carboxypropylamino)purine dinucleotide, a functionalized NAD+ analogue carrying an omega-carboxyalkyl side-chain at the exocyclic adenine amino group. Carbodiimide coupling of the latter derivative to high-molecular-weight water-soluble (polyethyleneimine, polylysine) and insoluble (aminohexyl-Sepharose) polymers gave the corresponding macromolecularized NAD+ analogues. These derivatives have been shown to be enzymically reducible. The polyethyleneimine and polylysine analogues showed a substantial degree of efficiency relative to free NAD+ with rabbit muscle lactate dehydrogenase (60 and 25% respectively) but a lower one with yeast alcohol dehydrogenase and Bacillus subtilis alanine dehydrogenase (2-7%). The polyethyleneimine derivative entrapped in cellulose triacetate fibres together with the lactate dehydrogenase was operationally stable during repetitive use.
用3,4-环氧丁酸对NAD⁺腺嘌呤环的N-1位进行烷基化，随后化学还原为碱稳定的NADH形式并进行碱性迪姆罗思重排，得到在外环腺嘌呤氨基处烷基化的NADH衍生物。后一种衍生物的酶促再氧化得到烟酰胺-6-(2-羟基-3-羧丙基氨基)嘌呤二核苷酸，这是一种在外环腺嘌呤氨基处带有ω-羧基烷基侧链的功能化NAD⁺类似物。后一种衍生物与高分子量水溶性（聚乙烯亚胺、聚赖氨酸）和不溶性（氨基己基-琼脂糖）聚合物进行碳二亚胺偶联，得到相应的大分子化NAD⁺类似物。这些衍生物已被证明可被酶还原。聚乙烯亚胺和聚赖氨酸类似物相对于游离NAD⁺对兔肌肉乳酸脱氢酶显示出相当程度的效率（分别为60%和25%），但对酵母醇脱氢酶和枯草芽孢杆菌丙氨酸脱氢酶的效率较低（2-7%）。与乳酸脱氢酶一起包埋在三醋酸纤维素纤维中的聚乙烯亚胺衍生物在重复使用过程中操作稳定。
