We first set up a blank to zero out the spectrophotometer. In tubes 1-4 we added 200 ul of DPIP and 200 ul of the bean extract enzyme, but in tube two, the bean extract was boiled. We then added the appropriated amount of phosphate buffer to the tubes. 200 ul of the inhibitor, malonate, was added to tube three and four. 66 ul of the substrate, succinate, was added to test tube one, two, and three but 200 ul of succinate was added to tube four. Tube one was our positive control. The enzyme was fully efficient and there was no inhibitor. It is expected that tube one will be catalyzed the quickest. Tube two was our negative control. We needed to observe what would occur in the spectrophotometer if the enzyme couldn’t catalyze the reaction and so we added a denatured enzyme to view the effects of no reaction. I expect that the reaction will occur the quickest for tube one, the slowest for tube two, and I expect that tube four will be quicker than tube
We first set up a blank to zero out the spectrophotometer. In tubes 1-4 we added 200 ul of DPIP and 200 ul of the bean extract enzyme, but in tube two, the bean extract was boiled. We then added the appropriated amount of phosphate buffer to the tubes. 200 ul of the inhibitor, malonate, was added to tube three and four. 66 ul of the substrate, succinate, was added to test tube one, two, and three but 200 ul of succinate was added to tube four. Tube one was our positive control. The enzyme was fully efficient and there was no inhibitor. It is expected that tube one will be catalyzed the quickest. Tube two was our negative control. We needed to observe what would occur in the spectrophotometer if the enzyme couldn’t catalyze the reaction and so we added a denatured enzyme to view the effects of no reaction. I expect that the reaction will occur the quickest for tube one, the slowest for tube two, and I expect that tube four will be quicker than tube