METABOLIC FLUX ANALYSIS OF PICHIA STIPITIS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 我们的目标是测量稳定状态下的细胞内代谢通量，这将提供关于细胞新陈代谢的定量信息，并使我们能够确定可能的基因靶点以进行菌株改进。将~(13)C标记的葡萄糖喂入细胞，酵母细胞用6M盐酸水解液冷冻干燥。氨基酸的同位异构体组成将使用HSQC进行测量，其中将使用最强的碳信号进行积分。在输入碳源的基础上，通过最小化测量数据和模拟数据来模拟碳代谢通量的分布。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Our objective is to measure the intracellular metabolic flux under steady state; this will provide quantitative information about cell metabolism and allows us to identify possible gene targets for strain improvement. 13C labeled glucose is fed to the cell; yeast cell will be hydrolyzed using 6M HCl and lyophilized. Isotopmer composition of amino acid will be measured using HSQC, in which a slice of strongest carbon signal will be used for integration. The carbon metabolic flux distribution will be simulated by minimizing measured data and simulated data based on input carbon source.