METABOLIC FLUX ANALYSIS OF PICHIA STIPITIS

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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.