31P nuclear magnetic resonance studies of the fermentation of glucose to ethanol by Zymomonas mobilis

K. D. Barrow, J. G. Collins, R. S. Norton

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Abstract

High resolution 31P nuclear magnetic resonance spectroscopy has been employed to study the fermentation of glucose to ethanol by Zymomonas mobilis, strain ZM4, a bacterium which uses the Entner-Doudoroff pathway. The levels of nucleoside triphosphates, sugar phosphates, UDP-sugars and P(i) in intact fermenting cells have been studied with a time resolution of 1 min. It is suggested that a pH gradient is established across the cell membrane during fermentation and that the intracellular pH does not rise above approximately 6.4. 31P resonances from most phosphorus-containing intermediates in the Entner-Doudoroff pathway, as well as adenosine and uridine nucleotides and a number of other intracellular metabolites, have been assigned in perchloric extracts of fermenting cells by means of a number of techniques, including two-dimensional homonuclear J-resolved and two-dimensional homonuclear shift-correlated spectroscopy. Quantification of these metabolites in spectra of extracts of fermenting cells indicates that the rate-limiting steps in the Entner-Doudoroff pathway in Z. mobilis are the conversions of glucose 6-phosphate to 6-phosphogluconate and of 3-phosphoglycerate to 2-phosphoglycerate.

Original languageEnglish
Pages (from-to)5711-5716
Number of pages6
JournalThe Journal of Biological Chemistry
Volume259
Issue number9
Publication statusPublished - 1984
Externally publishedYes

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