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An enzyme is a protein molecule that can create chemical reactions in a much quicker rate inside cells. People create enzymes in their body which is called proteins. The structure of an enzyme is a connection of amino acids. These amino acids have a very specific sequence that allows them to connect together to make a shape. The cell’s nucleus determines the sequences because of its genes. Enzymes have an active site that allows substances to connect to it to create a chemical reaction. This creates a new shape with the enzyme and the outcome is the product which leaves to interact with other substances. For enzymes to function they need cofactors or coenzymes. Cofactors can be metal and coenzymes can be vitamins. The reason for enzymes existence is to create chemical reactions to change things. For example, if a person wants to lose weight, they have to put in the work to get there just like enzymes; they put in the work to create a change.
Furthermore, there are items that can affect how an enzyme functions. An example of this is temperature. Humans work differently at different temperatures whether outside temperature or body temperature. For example, if it is cold outside, people could work slower due them shivering, but if it’s hot, they might work more tiresome due to heat. Same goes for enzymes because the hotter it gets, the less it can keep forming, but the colder it is, the less work it does. Enzymes have a specific temperature that it works well at. The proper term for this is denature, which means that at a certain point, the enzyme will no longer keep forming and therefore stay constant.
The experiments were used to measure the enzyme reaction at different dilutions and temperatures. Horseradish was used as the peroxidase because it would calculate the oxidation by hydrogen peroxide. Also, guaiacol was used to measure the reaction by the color of the reaction mixture. The color would be a brownish color and the spectrophotometer would measure it precisely. The effect of enzyme concentration on the rate of reaction is that when the enzyme concentration doubles, the rate of reaction doubles as well. If true, then the enzyme concentration reaction will keep increasing with each test.
The temperature on rate of reaction experiment involved 4 different temperatures. They were 5 degrees celsius, 21 degrees celsius, 34 degrees celsius, and 60 degree celsius. These are cold temperature, room temperature, warm temperature, and boiling hot temperature. As the temperature goes from cold to boiling temperature, the absorbance will increase drastically and therefore, the rate will be higher too. If true, then as the temperature increases with each test, the absorbance will too.
Material and Method:
Students in General Biology 1 did a lab based on the effects of enzyme concentration on absorbance. The students began by having to make an enzyme extract which was a mixture of a horseradish that weighed 1 gram, 100 milliliters of citrate-phosphate buffer, and it was blended for 15 seconds. They used a cheesecloth to cover a 100 milliliter beaker so the leftovers of the horseradish did not enter it, then poured the mixture into the beaker, and labeled it ‘enzyme extract.’
Next, another 100 milliliter beaker was labeled ‘buffer.’ This beaker contained 25 milliliters of citrate-phosphate ph 5.0, and two dispensers of H202 and guaiacol were placed on a table. With the solutions, buffers, and enzyme extract, they filled nine test tubes using the chart describing how much of each to put into the test tubes.
Then, students prepared a spectrophotometer, which is a machine to measure absorbance. The spectrophotometer was set at 500 wavelengths and was blanked with test tube number one. The students mixed test tubes 2 and 3 together, 4 and 5 together, 6 and 7 together, 8 and 9 together because they were the different dilutions. Each mixture was mixed and placed into the spectrophotometer and recorded at every 20 second interval up to 120 seconds. Then, the spectrophotometer was blanked and the next mixture would be absorbed.
Moreover, in the second experiment, students learned the effects of different temperatures on the rate of reaction. Students used the same enzyme extract from the previous experiment and then students choose the dilution with the highest initial increase without it denaturing. A group choose the 1.0 milliliter dilution and began the experiment.
Additionally, students obtained nine test tubes to place the buffer, enzyme extract, and dilution in. Test tubes 3, 5, 7, and 9 had the same volumes of H2O2 and guaiacol. Test tubes 2, 4, 6, and 8 had the same volumes of buffer and enzyme extract. Test tube 1 contained buffer, H2O2, and guaiacol. This was used as the blank for the spectrophotometer. Test tubes 2 and 3 were placed in ice water at 5 degree celsius. Test tubes 4 and 5 were kept at room temperature which was 21 degrees celsius. Test tubes 6 and 7 were placed in a 34 degree celsius incubator. Test tubes 8 and 9 were placed in a 60 celsius incubator. All the test tube were in those temperatures for 10 minutes but each couplet were put one at a time.
Then, the spectrophotometer was calibrated to 500 wavelengths and was blanket by test tube one. After each couplet were finished with the 10 minute period, they were mixed and were placed in the spectrophotometer. The students recorded the absorbance for 20 second interval for 120 seconds.

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