D-[14C]核酮糖1,5-二磷酸的制备规模酶促合成。
Preparative-scale enzymic synthesis of D-[14C]ribulose 1,5-bisphosphate.
作者信息
Kuehn G D, Hsu T C
出版信息
Biochem J. 1978 Dec 1;175(3):909-12. doi: 10.1042/bj1750909.
Abstract
A procedure is described to prepare uniformly labelled D-[14C]ribulose 1,5-bisphosphate enzymically from uniformly labelled D-[14C]glucose through the coupled reactions catalysed by hexokinase (EC 2.7.1.1), glucose 6-phosphate dehydrogenase (EC 1.1.1.49), 6-phosphogluconate dehydrogenase (EC 1.1.1.44) and 5-phosphoribulokinase (EC 2.7.1.19). All reagents utilized in the method are commercially available. The procedure is a reliable preparative-scale method for synthesizing the dibarium salt of D-[14C]ribulose 1,5-biphosphate with a specific radioactivity up to 7 mCi/mmol and a purity near 90%. The final product was free of other 14C-labelled sugars, sugar phosphate esters, Pi and nucleotides.
摘要
描述了一种通过己糖激酶（EC 2.7.1.1）、葡萄糖6-磷酸脱氢酶（EC 1.1.1.49）、6-磷酸葡萄糖酸脱氢酶（EC 1.1.1.44）和5-磷酸核酮糖激酶（EC 2.7.1.19）催化的偶联反应，从均匀标记的D-[14C]葡萄糖酶法制备均匀标记的D-[14C]核酮糖1,5-二磷酸的方法。该方法中使用的所有试剂均可商购获得。该方法是一种可靠的制备规模方法，用于合成比活度高达7 mCi/mmol且纯度接近90%的D-[14C]核酮糖1,5-二磷酸钡盐。最终产物不含其他14C标记的糖、糖磷酸酯、无机磷酸和核苷酸。
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