Metabolic engineering applications of in vivo sup 31 P and sup 13 C NMR studies of Saccharomyces cerevisiae
With intent to quantify NMR measurements as much as possible, analysis techniques of the in vivo {sup 31}P NMR spectrum are developed. A systematic procedure is formulated for estimating the relative intracellular concentrations of the sugar phosphates in S. cerevisiae from the {sup 31}P NMR spectrum. In addition, in vivo correlation of inorganic phosphate chemical shift with the chemical shifts of 3-phosphoglycerate, {beta}-fructose 1,6-diphosphate, fructose 6-phosphate, and glucose 6-phosphate are determined. Also, a method was developed for elucidation of the cytoplasmic and vacuolar components of inorganic phosphate in the {sup 31}P NMR spectrum of S. cerevisiae. An in vivo correlation relating the inorganic phosphate chemical shift of the vacuole with the chemical shift of the resonance for pyrophosphate and the terminal phosphate of polyphosphate (PP{sub 1}) is established. Transient measurements provided by {sup 31}P NMR are applied to reg1 mutant and standard strains. {sup 31}P and {sup 13}C NMR measurements are used to analyze the performance of recombinant strains in which the glucose phosphorylation step had been altered.
- Research Organization:
- California Inst. of Tech., Pasadena, CA (USA)
- OSTI ID:
- 7255399
- Resource Relation:
- Other Information: Thesis (Ph. D.)
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
GLUCOSE
PHOSPHORYLATION
PHOSPHATES
BIOSYNTHESIS
SACCHAROMYCES CEREVISIAE
NMR SPECTRA
CARBON 13
CHEMICAL SHIFT
IN VIVO
NUCLEAR MAGNETIC RESONANCE
PHOSPHORUS 31
TRACER TECHNIQUES
ALDEHYDES
CARBOHYDRATES
CARBON ISOTOPES
CHEMICAL REACTIONS
EUMYCOTA
EVEN-ODD NUCLEI
FUNGI
HEXOSES
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
MAGNETIC RESONANCE
MICROORGANISMS
MONOSACCHARIDES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
OXYGEN COMPOUNDS
PHOSPHORUS COMPOUNDS
PHOSPHORUS ISOTOPES
PLANTS
RESONANCE
SACCHARIDES
SACCHAROMYCES
SPECTRA
STABLE ISOTOPES
SYNTHESIS
YEASTS
550501* - Metabolism- Tracer Techniques