Mathematical modelling The form of the Hill equation was: = [S]n/(Kbn+[S]n) (1) v Vmax where is the maximal velocity, [S] the substrate concentration, the apparent Vmax Kb dissociation binding constant, the Hill coefficient (measure of enzyme n cooperativity) and the velocity (i.e., rate of reaction) at the given substrate v concentration [S]. The fitting was initially implemented in Microsoft Excel 2010 and, in a more refined form, in Wolfram using the inbuilt Mathematica routine. This delivered the best-fit values for kinetic parameters ‘NonlinearModelFit’ along with their standard errors. NMR spectroscopy 3.56 mL aliquots of 10 mM [1,1,1-13C]triolein emulsions were mixed with 360 of μL D2O (for field frequency locking). 40 of 1 M sodium [2-13C]acetate in D2O and μL 100 of 180 mM NaHPO4 were added as internal references for 13C-NMR and μL 31P-NMR spectra, respectively. The samples were re-sonicated and 3.5 mL of each emulsion was transferred into a 10-mm glass NMR tube (Wilmad, Buena, NJ, USA). The pH of the NMR samples was maintained at 8.0 ± 0.1 and the temperature of the samples was set to 35°C. 31P-NMR and 13C-NMR spectra were recorded from the same emulsions using a 9.4 T vertical-bore magnet (Oxford Instruments, Oxford, UK), equipped with Avance III 400 console (Bruker, Karlsruhe, Germany) and a Bruker broadband observe (BBO) probe. The excitation-pulse angle was ~30˚ and inverse-gated proton WALTZ-16 decoupling was used during acquisition to eliminate splitting due to 1H-31P or 1H-13C with the supplied Bruker J-couplings,
Mathematical modelling The form of the Hill equation was: v = Vmax [S]n/(Kbn+[S]n) (1) where Vmax is the maximal velocity, [S] the substrate concentration, Kb the apparent dissociation binding constant, n the Hill coefficient (measure of enzyme cooperativity) and v the velocity (i.e., rate of reaction) at the given substrate concentration [S]. The fitting was initially implemented in Microsoft Excel 2010 and, in a more refined form, in Wolfram Mathematica using the inbuilt ‘NonlinearModelFit’ routine. This delivered the best-fit values for kinetic parameters along with their standard errors. NMR spectroscopy 3.56 mL aliquots of 10 mM [1,1,1-13C]triolein emulsions were mixed with 360 μL of D2O (for field frequency locking). 40 μL of 1 M sodium [2-13C]acetate in D2O and 100 μL of 180 mM NaHPO4 were added as internal references for 13C-NMR and 31P-NMR spectra, respectively. The samples were re-sonicated and 3.5 mL of each emulsion was transferred into a 10-mm glass NMR tube (Wilmad, Buena, NJ, USA). The pH of the NMR samples was maintained at 8.0 ± 0.1 and the temperature of the samples was set to 35°C. 31P-NMR and 13C-NMR spectra were recorded from the same emulsions using a 9.4 T vertical-bore magnet (Oxford Instruments, Oxford, UK), equipped with Avance III 400 console (Bruker, Karlsruhe, Germany) and a Bruker broadband observe (BBO) probe. The excitation-pulse angle was ~30˚ and inverse-gated proton WALTZ-16 decoupling was used during acquisition to eliminate splitting due to 1H-31P or 1H-13C J-couplings, with the supplied Bruker