Dihydroflavonol 4-reductase (DFR), a member of the short-chain dehydrogenase family, catalyzes the last common step in the biosynthesis of flavan-3-ols and condensed tannins. Initial rates of DFR were measured by moni...Dihydroflavonol 4-reductase (DFR), a member of the short-chain dehydrogenase family, catalyzes the last common step in the biosynthesis of flavan-3-ols and condensed tannins. Initial rates of DFR were measured by monitoring the 340-nm absorbance decrease resulting from the joint consumption of dihydroquercetin (DHQ) and NADPH, as a function of pH, temperature and ionic strength. At pH 6.5 and 30o C, substrate inhibition was observed above 30 μM DHQ. At lower/non-inhibitory DHQ concentrations, NADP+ behaves as a competitive inhibitor with respect to NADPH and as a mixed inhibitor with respect to DHQ, which supports a sequential ordered mechanism, with NADPH binding first and NADP+ released last. Binding-equilib-rium data obtained by means of the chromatographic method of Hummel and Dreyer at pH 7.5 and by isothermal calorimetric titration at pH 6.5 led to the conclusion that ligands of the apoenzyme included NADPH, NADP+ and DHQ. The mechanism which best accounts for substrate inhibition at pH 6.5 in the absence of product involves the formation of a binary non-productive E.DHQ complex. Thus, a productive ternary complex cannot be formed when DHQ binds first. This mechanism of inhibition may prevent the accumulation of unstable leucoanthocyanidins within cells.展开更多
A new weak boundary procedure for hyperbolic problems is presented.We consider high order finite difference operators of summation-by-parts form with weak boundary conditions and generalize that technique.The new boun...A new weak boundary procedure for hyperbolic problems is presented.We consider high order finite difference operators of summation-by-parts form with weak boundary conditions and generalize that technique.The new boundary procedure is applied near boundaries in an extended domain where data is known.We show how to raise the order of accuracy of the scheme,how to modify the spectrum of the resulting operator and how to construct non-reflecting properties at the boundaries.The new boundary procedure is cheap,easy to implement and suitable for all numerical methods,not only finite difference methods,that employ weak boundary conditions.Numerical results that corroborate the analysis are presented.展开更多
文摘Dihydroflavonol 4-reductase (DFR), a member of the short-chain dehydrogenase family, catalyzes the last common step in the biosynthesis of flavan-3-ols and condensed tannins. Initial rates of DFR were measured by monitoring the 340-nm absorbance decrease resulting from the joint consumption of dihydroquercetin (DHQ) and NADPH, as a function of pH, temperature and ionic strength. At pH 6.5 and 30o C, substrate inhibition was observed above 30 μM DHQ. At lower/non-inhibitory DHQ concentrations, NADP+ behaves as a competitive inhibitor with respect to NADPH and as a mixed inhibitor with respect to DHQ, which supports a sequential ordered mechanism, with NADPH binding first and NADP+ released last. Binding-equilib-rium data obtained by means of the chromatographic method of Hummel and Dreyer at pH 7.5 and by isothermal calorimetric titration at pH 6.5 led to the conclusion that ligands of the apoenzyme included NADPH, NADP+ and DHQ. The mechanism which best accounts for substrate inhibition at pH 6.5 in the absence of product involves the formation of a binary non-productive E.DHQ complex. Thus, a productive ternary complex cannot be formed when DHQ binds first. This mechanism of inhibition may prevent the accumulation of unstable leucoanthocyanidins within cells.
基金supported by the National Science Foundation under Award No.0948304 and by the Southern California Earthquake Center.SCEC is funded by NSF Cooperative Agreement EAR-0529922 and USGS Cooperative Agreement 07HQAG0008(SCEC contribution number 1806).The work by the last author was carried out within the Swedish e-science Research Centre(SeRC)and supported by the Swedish Research Council(VR).
文摘A new weak boundary procedure for hyperbolic problems is presented.We consider high order finite difference operators of summation-by-parts form with weak boundary conditions and generalize that technique.The new boundary procedure is applied near boundaries in an extended domain where data is known.We show how to raise the order of accuracy of the scheme,how to modify the spectrum of the resulting operator and how to construct non-reflecting properties at the boundaries.The new boundary procedure is cheap,easy to implement and suitable for all numerical methods,not only finite difference methods,that employ weak boundary conditions.Numerical results that corroborate the analysis are presented.
