Methods for the enzymic synthesis, isolation, purification and analysis of the 14C labelled intermediates that are characteristic of the L-type pentose phosphate pathway are described. These are D-glycero D-ido octulose 1, 8-bisphosphate and 8-monophosphate; D-glycero D-altro octulose mono- and bisphosphates; sedoheptulose 1,7-bisphosphate and D-arabinose 5-phosphate. The procedures for the preparation of octulose- and sedoheptulose bisphosphates are based on the aldolase-catalyzed condensation of dihydroxyacetone phosphate with appropriately 14C-labelled aldo pentose 5-phosphates and erythrose 4-phosphate respectively. D-Glycero D-ido octulose 8-phosphate and its altro epimer were prepared in transketolase catalyzed reactions involving β-hydroxypyruvate and appropriately 14C-labelled aldohexose 6-phosphates. [U-14C] D-arabinose 5-phosphate was synthesized by reacting [U-14C] glucosamine 6-phosphate with ninhydrin. The authenticity of each of the 14C-labelled sugar phosphates was confirmed using a variety of chromatographic methods, enzymatic analytic methods and NMR spectroscopy. The 14C labelled compounds are used in investigations involving the search for the identity of the pentose pathway in tissues invitro, for the measurement of L-type pathway enzyme reactions and for testing the mechanistic predictions of the L-type pathway in vitro.

中文翻译：

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L 型戊糖途径的 14C 标记中间体的高产合成：单磷酸和双磷酸辛酮糖、1,7-双磷酸景天酮糖和 D-阿拉伯糖 5-磷酸

描述了作为 L 型磷酸戊糖途径特征的 14C 标记中间体的酶合成、分离、纯化和分析方法。它们是 D-甘油 D-碘辛酮糖 1、8-二磷酸和 8-单磷酸；D-甘油 D-altro octulose 单磷酸和双磷酸；景天庚酮糖 1,7-二磷酸和 D-阿拉伯糖 5-磷酸。制备二磷酸八酮糖和景天庚酮糖的程序基于二羟基丙酮磷酸分别与适当的 14C 标记的醛戊糖 5-磷酸和赤藓糖 4-磷酸的醛缩酶催化缩合。D-Glycero D-ido octulose 8-phosphate 及其异构差向异构体是在转酮醇酶催化的反应中制备的，该反应涉及 β-羟基丙酮酸和适当的 14C 标记的 6-磷酸己醛糖。[U-14C] D-阿拉伯糖 5-磷酸是通过 [U-14C] 氨基葡萄糖 6-磷酸与茚三酮反应合成的。使用各种色谱方法、酶分析方法和核磁共振光谱法证实了每种 14C 标记的糖磷酸盐的真实性。14C 标记的化合物用于研究，包括在体外组织中寻找戊糖途径的身份，用于测量 L 型途径酶反应以及在体外测试 L 型途径的机械预测。