Römisch Werner, Eisenreich Wolfgang, Richter Gerald, Bacher Adelbert
Lehrstuhl für Organische Chemie und Biochemie, Technische Universität München, Lichtenbergstr. 4, D-85747 Garching, Germany.
J Org Chem. 2002 Dec 13;67(25):8890-4. doi: 10.1021/jo026105x.
Flavocoenzymes labeled with stable isotopes are important reagents for the study of flavoproteins using isotope-sensitive methods such as NMR, ENDOR, infrared, and Raman spectroscopy. We describe highly versatile one-pot methods for the preparation of riboflavin isotopomers labeled with (13)C in every desired position of the xylene moiety. The starting materials are commercially available (13)C-labeled glucose samples, which are converted into riboflavin using enzymes of the oxidative pentose phosphate pathway in combination with recombinant enzymes of the riboflavin biosynthetic pathway. The overall reaction comprises six enzyme-catalyzed reaction steps for the synthesis of the vitamin and two auxiliary enzymes for in situ recycling of cofactors. The overall yields of riboflavin based on isotope-labeled glucose are 35-50%.
用稳定同位素标记的黄素辅酶是使用诸如核磁共振（NMR）、电子核双共振（ENDOR）、红外和拉曼光谱等对同位素敏感的方法研究黄素蛋白的重要试剂。我们描述了高度通用的一锅法，用于制备在二甲苯部分的每个所需位置都用¹³C标记的核黄素同位素异构体。起始原料是市售的¹³C标记的葡萄糖样品，其使用氧化戊糖磷酸途径的酶与核黄素生物合成途径的重组酶结合转化为核黄素。整个反应包括合成维生素的六个酶催化反应步骤和用于辅因子原位循环的两种辅助酶。基于同位素标记葡萄糖的核黄素总产率为35 - 50%。
