Changes in pH change the shape of the substrate so that it cannot bind with the active site or be catcalled, thus a product is produced more quickly due to higher substrate concentration until the point of the enzyme saturation is reached (Freeman, et al. , 2011). These concepts related to this experiment because as higher concentrations of pH were added to solution, the rate of hydrolysis of PUMP increased as well. The hypothesis stated that if the concentration of pH was increased, the rate of hydrolysis would increase as well, but if the concentration of pH was decreased, the rate of hydrolysis should decrease.
When performing this lab, the concentration of Citrate buffer was increased from a pH of 4. 8 M to 6. 0 M. By doing this, the rate of hydrolysis increased, therefore the data did support the hypothesis. The hypothesis also stated that once the reaction has reached its optimum pH, the reaction rate should slowly start decreasing. This was proved by the data because in the first experiment, the pH of the Citrate buffer was 4. 8 M and the rate of hydrolysis continuously increased, reaching 5. Xx-7 M/minute.
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In experiment two, the pH of the buffer was increased to 6. 0 M and the data fluctuated in rate. Thus, the hypothesis goes not need to be revised because it was correct and it was supported by the data. Moreover, the experiment does not need to be revised either, because it was done correctly and the results were around where they were supposed to be. Since the results support the hypothesis, it can be assumed that the increase of pH has a significant relationship with the rate of hydrolysis in that the increase of one will increase the other, and vice versa.
This would also suggest that if the pH of the Citrate buffer was to continue increasing to a very high value, the enzyme would become denatured, losing its shape and function, and therefore come useless in the hydrolysis reaction. A basis for this reasoning is that when the pH was increased from 4. 8 M to 6. 0 M, the reaction had already reached a point at which it started slowing down, showing a decrease in reaction rate from having too high of a pH level. From what is stated in the Introduction, the results from this experiment agree with the literature.
With an increase of pH, more enzymes are available to fit with the substrate and form enzyme substrate complexes, forming more products (Freeman, et al. , 2011). This is seen in the results as more PUMP is seen being produced as pH gets increased. If the concentration keeps increasing, there will be a point where the rate then becomes constant (Freeman, et al. , 2011). This is also seen in the data between test tubes three and four when the rate of hydrolysis drops from 1. Xx – 6 M/ min to 1. Ex. -6 M/min. This is a result of the reaction reaching the optimum pH concentration and denaturing the enzyme. The importance of the results from this experiment being accurate certifies that the hypothesis was correct, and the experimenter followed the procedure well. It clarified the relationship between pH and the rate of hydrolysis of PUMP. It also allows the experimenter to realize where they went wrong and to understand the effect of PUMP concentration on the hydrolysis of PUMP.
There were no significant errors in the procedure of this experiment that changed the results drastically; however there were minor observational, systematic and instrumental errors. Throughout this experiment, many minor errors could have altered the results of the trials. To begin, the experiment included multiple trials of petting that could have been off due to instrumental and systematic errors. Since petting is not always accurate, there ay have been more or less substrate, Citrate buffer, enzyme or water added to each test tube.
Another form of instrumental error COUld be the absorbency read of the spectrophotometer. The suspects may not have been cleaned thoroughly, or the results may just be off. However, despite these errors, the hypothesis was still correct that the rate of hydrolysis increased as the pH increased, but the numbers may have been off. Throughout the experiment, the directions were followed well. In the future, being organized is an asset because petting will not be rushed, and the systematic errors will be reduced, giving more accurate results.