That enzymes bind with chemical reactants called substrates. There may be one or more substrates for each type of enzyme, depending on the particular chemical reaction. . In this experiment, different amount of enzyme and substrate were put in a test tube,then it differ in speed capacity. The faster the velocity of the reaction, the sooner half of Vmax is reached the smaller the value of Km. If the enzyme have high substrate affinity, it will bind its target even at low concentration(low Km).
In experiment; we measure the enzyme activity, in this experiment enzyme that we used is LDH (H4) that catalyzes the conversion of L-lactate and NAD+ to become pyruvate and NADH. The process of reaction we can observed by the spectrometer with the value recorded every 10 seconds for 3 minutes. The results can be used to draw a linear graph of OD340nm(Y-axis) and mol of NADH(x-axis) and calculate the value of Km and Vmax from the interception point in the graph. In the first experiment It was found that as the concentration of enzyme was increased, the speed of reaction increased. Likewise, it was found that as the concentration of substrate was increased, the speed of reaction increased;minor errors in the experiments are caused by incorrect pipetting of the solutions. In second experiment we assay the enzymatic activities of LDH undee excessive(saturated) amount of substrates and products. In Third experiment we used 4 tubes with different concentration of sodium lactate (30mM, 20mM, 10mM and 5mM) the result are the aame with previous experiments.slight error in the experiments are caused by forgot to kept the enzyme solution on ice,which would become worse.