José Manuel Horcas Nieto

161 5 Establishing a peroxisomal β-oxidation computational kinetic model to understand the effects of amino-acid restriction Volume of peroxisome 1.8·10-6 L·mgProtein 3 NAD 250 µM 3 NADH 25 µM 3 AcetylCoA 700 µM 3 FreeCoA 700 µM 14 Carnitine 2000 µM 7 Oxygen 260 µM / Assumptions and calculations The Km of ACOX1 for H2O2 was estimated to be higher than that of oxygen as the enzyme runs in the forward direction and it is not expected to have high affinity for the product. No Km value was found in the literature. The value 10 is theoretical and not expected to affect the reaction, given the high Keq. The Vmax of CRAT used was the same as found for CROT as no studies reporting the absolute value were found. Km_CoA of CRAT was estimated by averaging the values of Km for Coa in the presence of C6, C8 and C10 of CROT. The Vmax of Catalase was calculated by averaging the Vmax of different papers isolated from different organisms. The average obtained was 46249.99, therefore we approximated it to 46000. The concentration of oxygen was estimated to be the solubility of oxygen in water at 37 °C. When expressed per peroxisomal protein this volumes was assumed to be similar to that of mitochondria. This assumption is not so strange as it seems, since the volume per protein is similar for mitochondria and cytosol (value X versus value Y), suggesting that its order of magnitude is not very different for different cellular compartments.

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