Illarionov Boris, Zhu Feng, Eisenreich Wolfgang, Bacher Adelbert, Weber Stefan, Fischer Markus
Hamburg School of Food Science, University of Hamburg , Grindelallee 117, 20146 Hamburg, Germany.
J Org Chem. 2015 Mar 6;80(5):2539-44. doi: 10.1021/jo502480w. Epub 2015 Feb 24.
Isotope-labeled flavins are crucial reporters for many biophysical studies of flavoproteins. A purine-deficient Escherichia coli strain engineered for expression of the ribAGH genes of Bacillus subtilis converts isotope-labeled purine supplements into the riboflavin precursor, 6,7-dimethyl-8-ribityllumazine, with yields up to 40%. The fermentation products can subsequently be converted into isotope-labeled riboflavin and the cognate flavocoenzymes, FMN and FAD, by in vitro biotransformation with better than 90% yield. Using this approach, more than 100 single or multiple (13)C-, (15)N-, (17)O-, and (18)O-labeled isotopologues of these cofactors and ligands become easily accessible, enabling advanced ligand-based spectroscopy of flavoproteins and lumazine receptor proteins at unprecedented resolution.
同位素标记的黄素是许多黄素蛋白生物物理研究的关键报告分子。一种经过工程改造用于表达枯草芽孢杆菌ribAGH基因的嘌呤缺陷型大肠杆菌菌株，能将同位素标记的嘌呤补充物转化为核黄素前体6,7-二甲基-8-核糖基卢马嗪，产率高达40%。随后，通过体外生物转化，发酵产物可转化为同位素标记的核黄素以及同源黄素辅酶FMN和FAD，产率高于90%。利用这种方法，这些辅因子和配体的100多种单标记或多标记（13）C-、（15）N-、（17）O-和（18）O-同位素类似物变得易于获取，从而能够以前所未有的分辨率对黄素蛋白和卢马嗪受体蛋白进行先进的基于配体的光谱分析。
