NEW PATHWAY OF ISOPRENOID BIOSYNTHESIS

SIR提供了(2S)-N-(2‘-O-benzyl-glycoloyl)bomane-10-，2-sultam [11，2-13C]乙酸；20G最近，合成异戊烯的新途径 已发现焦磷酸盐来自碳水化合物 新陈代谢。探索这一新途径在自然界中的分布和 为了进一步阐明该途径中的中间产物，我们目前正在 1-脱氧-D-[2，4-13C2]木糖的合成。最初，一个小的 勘探工作的数量将从乙基溴化物-[I- I 3C]乙酸酯和[2-13C]丙酮，这是由UW开发的 一群人。在这项工作的过程中，很明显，这条路线 效率不是很高，因此不适合生产更大的 这种前体的数量。因此，第二条路线是 由SIR设计，它有效地利用了标记的合成子 他们已经发展起来了。如果在E. 与最初合成的前体样本进行比较，证明 这种方法的可行性，大量的化合物将 然后由先生来综合。这一先驱物将被馈送到 合成异戊二烯类化合物的各种生物系统和 产物将通过~(13)C-核磁共振进行分析，以确定是否含有~(13)C 以及13C_13C偶合的出现，这是由于 前体的重新排列，在适当的位置。这 将概述新途径的相对重要性 与经典的甲氧戊酸途径相比。接下来，我们将孵化 洗涤细胞和无细胞提取物的双标记底物 直接在核磁共振光谱仪中检测到 通过~1H-检测~(13)C-核磁共振定期孵化，特别是 对于双量子跃迁，对于新物种的出现 这两个13C原子核相互连接。因为虚拟的 没有其他代谢途径可以引起广泛的13C-13C 耦合，这将是一个非常特定的探针，用于中间体 类异戊二烯生物合成的新途径。时间进程的时间进程 新物种的出现和消失将表明他们的 在反应序列中的相对位置，以及标记 更多职位的前身将允许分配 这些中间体的结构。这些活体实验将 需要很高的灵敏度才能探测到来自 少量的暂态化合物。这可以在750上实现 大学的兆赫核磁共振波谱仪。沃什。

The SIR provided (2S)-N-(2'-O-benzyl-glycoloyl)bomane-10-,2-sultam 11,2- 13C]acetic acid; 20g Recently, a new pathway to isopentenyl pyrophosphate has been discovered which emanates from carbohydrate metabolism. To probe distribution of this new pathway in nature and to elucidate further intermediates in the pathway, we are presently synthesizing 1-deoxy-D-[2,4-13C2]xylulose. Initially, a small quantity for exploratory work will be prepared from ethyl bromo- [ I - I 3C] acetate and [2-13C]acetone, which has been developed by the UW group. In the course of this work it became evident that this route is not very efficient and thus unsuitable for the production of larger quantities of this precursor. A second route has therefore been designed by the SIR, which makes effective use of the labeled synthons they have developed. If experiments on ubiquinone formation in E. coli with the initially synthesized precursor sample demonstrate the feasibility of this approach, a larger quantity of the compound will then be synthsized by the SIR. This precursor will be fed to a variety of biological systems synthesizing isoprenoid compounds and the products will be analyzed by 13C-NMR for the incorporation of 13C and the appearance of 13C_13C coupling, resulting from the rearrangement of the precursor, in the appropriate positions. This will give an overview of the relative importance of the new pathway compared to the classical mevalonate pathway. Next, we will incubate the double labeled substrate with washed cells and cell-free extracts of E. coli directly in the NMR spectrometer and examine the incubations periodically by 1H-detected 13C-NMR, looking specifically for double-quantum transitions, for the appearance of new species in which the two 13C nuclei are connected to each other. Since virtually no other metabolic pathway can give rise to extensive 13C-13C coupling, this will be a very specific probe for the intermediates in the new pathway of isoprenoid biosynthesis. The time course of the appearance and disappearance of new species will indicate their relative position in the reaction sequence, and labeling of the precursor in additional positions will allow the assignment of structures to these intermediates. These in vivo experiments will require high sensitivity to detect multiquantum transitions from very small amounts of transient compounds. This can be achieved on the 750 MHz NMR spectrometer at the Univ. of Wash.