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Comparison of wild‐type and Reg 1 mutant saccharomyces cerevisiae metabolic levels during glucose and galactose metabolism using 31 P NMR
Author(s) -
Shanks Jacqueline Vanni,
Bailey James E.
Publication year - 1990
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260350407
Subject(s) - fructose , biochemistry , glucose 6 phosphate , mutant , hexokinase , glycolysis , galactose , saccharomyces cerevisiae , metabolism , nuclear magnetic resonance spectroscopy , chemistry , wild type , strain (injury) , carbohydrate metabolism , biology , enzyme , gene , stereochemistry , anatomy
The reg 1 mutation will allow the expression of a cloned gene on a plasmid under the control of a GAL promoter in the presence of glucose. The metabolism of wild‐type and reg l mutant strains was examined by in vivo 31 P nuclear magnetic resonance (NMR) spectroscopy. Transient profiles of glucose 6‐phosphate, fructose 6‐phosphate, fructose 1, 6‐diphosphate, and 3‐phosphoglycerate indicated that glucose was processed differently for the reg 1 strain despite similar cytoplasrnic pH values and ATP levels. Intracellular phosphate became depleted in the transition to quasi‐steady state and limited glycolysis in the reg 1 strain. The glucose uptake step or hexokinase step appears to be altered in the reg 1 strain. The reg 1 strain utilized galactose faster than the wild‐type strain under the conditions used for NMR analysis. These results are consistent with the hypothesis that the REG 1 product operates early in the regulatory circuitry for glucose repression. This study illustrates the usefulness of transient information provided by NMR in understanding changes in the metabolism of genetically manipulated organisms.