Introduction: Biochemical pathways are essentially linked series of enzymatic reactions that are interdependent. Their activity is coordinated by sensitive means of communication. This laboratory experiment examines the kinetic parameters, Vmax and Km, of three enzymes including creatine phosphokinase, pyruvate kinase and lactate dehydrogenase. Two of the enzymes, pyruvate kinase and lactate dehydrogenase, make up the metabolic pathway of glycolysis, a sequence of reactions that metabolizes glucose into pyruvate with a net production of ATP. Each enzyme has two substrates; kinetic analysis was preformed for each substrate. The kinetic parameters for each enzyme were examined by monitoring the disappearance of NADH at 340 nm as the enzyme converted pyruvate to lactate. . NADH has maximum absorbance at 340nm and when oxidized (NAD+) there is a dramatic difference in the absorbance spectra. The kinetic analysis involved in this experiment is solely based on this difference in NADH absorbance over time. Although each enzyme has a different pathway leading to the production lactate, N......

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