Measurement of (carbon) kinetic isotope effect by Rayleigh fractionation using membrane inlet mass spectrometry for CO2 -consuming reactions

Functional Plant Biology : FPB
Dennis B McNevinGraham D Farquhar

Abstract

Methods for determining carbon isotope discrimination, Δ, or kinetic isotope effects, α, for CO2-consuming enzymes have traditionally been cumbersome and time-consuming, requiring careful isolation of substrates and products and conversion of these to CO2 for measurement of isotope ratio by mass spectrometry (MS). An equation originally derived by Rayleigh in 1896 has been used more recently to good effect as it only requires measurement of substrate concentrations and isotope ratios. For carboxylation reactions such as those catalysed by d-ribulose-1,5-bisphosphate carboxylase / oxygenase (RuBisCO, EC 4.1.1.39) and PEP carboxylase (PEPC, EC 4.1.1.31), this has still required sampling of reactions at various states of completion and conversion of all inorganic carbon to CO2, as well as determining the amount of substrate consumed. We introduce a new method of membrane inlet MS which can be used to continuously monitor individual CO2 isotope concentrations, rather than isotope ratio. This enables the use of a simplified, new formula for calculating kinetic isotope effects, based on the assumptions underlying the original Rayleigh fractionation equation and given by.

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May 1, 2005·Functional Plant Biology : FPB·Guillaume Tcherkez, Graham D Farquhar

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Citations

Jul 16, 2009·Rapid Communications in Mass Spectrometry : RCM·Christiane WernerPierrick Priault
Jun 28, 2012·Plant, Cell & Environment·Florian A BuschRowan F Sage
Aug 26, 2011·Journal of Molecular Modeling·Jan Philipp Götze, Peter Saalfrank
Aug 5, 2009·Photosynthesis Research·Katrin BeckmannWarwick Hillier
Nov 23, 2020·The New Phytologist·Pauliina Schiestl-AaltoAnnikki Mäkelä
Oct 17, 2008·Plant Physiology·Gary J LaniganUlli Seibt

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