Biology 1010 Enzyme
The presence of quinones prevents infection of microorganisms because it is toxic to them (Danyk, H, 2013). Quinones are produced by the oxidation of phenolic compound of catechol. Enzymes are used to speed up chemical reactions in cells (Danyk, H, 2013). The enzyme catechol oxidase is used to speed up the production of benzoquinone which is to help in infection prevention. In this study catechol oxidase was combined with potato juice extract and water. To provide more results of the enzymes productivity patterns the solutions; temperatures were changed, alterations were made in pH, and the amount of concentration of catechol was altered.
The solutions absorption was measured by a spectrophotmeter, this determined what conditions the enzyme produced benzoquinone the most. This is done to prove that the more basic the pH is, at room temperature, and with higher concentration of catechol oxidase it will produce more benzoquinone. This will happen until the present amount of catechol oxidase becomes limited due to its participation in the reaction. Methods and Materials There were six test tubes needed in the first procedure where the proper amount of mL was placed within; 1, 2, 4, 8, 16 and 24.
Biology 1010 Enzyme Essay Example
One at a time 5mL of the neutral pH 7 was added to the tubes where the equal amounts of catechol (corresponding the number on the tube) were also dropped in respectfully. To make sure that every test tube was equal in there amount of volume, more of the pH 7 were then added drop by drop, starting at the beginning; 23, 22, 20, 16, and 8. All consisting of equal parts in volume the test tubes were combined with 30 drops of potato juice extract from an eyedropper and shaken so the entire mixture could become equally distributed throughout the enclosed area.
Keeping the tubes at room temperature while mixing at one-minute intervals would continue to help the substrate availability have an effect on the enzyme activity. A 3-degree water bath would then set this activity so that the yield of enzymes catalyzed could be determined. In the occurrence of a pH change where any enzymes are present within the medium can cause the structures; both secondary and tertiary, to become disturbed which will cause these weak hydrogen bonds to become denatured. The extent of the denaturation of the bonded enzymes is affected by the emphasis of the pH change.
In cases where the pH change is very minimal there is the possibility that the correct structure can become regenerated completely from its optimum pH becoming reached again. Ideally this procedure will result in substances that are more basic and acidic to produce less benzoquinone and substances that have a pH of seven, or close to seven, will produce a higher concentration of benzoquinone because of the preferred neutrality that a pH of seven has. When preforming this procedure one must begin with five test tubes that are occupied with the correct amount of substance that has the pH of 4, 6, 7, 8, and 10 which will be used.
Once this buffer is created for each tube, 10 drops of potato juice as well as catechol oxidase containing the enzymes must be added and shaken in order to equally distribute all the components so that a possible change in pH can occur. Once placed in a 3 degree water bath the variety of colors in the substances due to the new pH change would have had a chance to completely set, making it possible to determine which pH is ideal for the most amount of benzoquinone to be produced. The amount of product is related to the temperature. The higher the temperature, the greater amount of collisions that will occur between the reactants.
This will then result in a higher amount of product. When the temperature reaches a specific point where it can no longer cause any further reactions, the weakest bonds will break causing the enzymes to undergo denaturation and lose their catalytic function. In order to complete this procedure six test tubes were retrieved where filled one by one with 3 mL each of pH 7. The test tubes were placed in their respective water baths (one each in a 3, 12, 20, 35, 50 and 70 degree water bath) while 10 drops of potato juice extract and catechol were added from an eyedropper, the tubes must stay sitting in their water baths during this step.
The tubes full of this mixture would be left for 15 minutes, stirring at one-minute intervals, so that they could ably reach the highest point that temperature is capable of and prohibit any further reactions to occur due to denaturation. Once this point had been reached the tubes could then be moved to a 3-degree water bath to continue to set so that the absorbance amount could then be determined from the effect that different temperatures have on catechol oxidase. Results The results presented are a mean of the class data. The lab manual presented information explaining the effects of catechol oxidase producing benzoquinone.
Seen, through the collected data more presence is seen absorbed in the solution when the concentration is higher (figure 1). With very little catechol added there was very little presence of benzoquinone making the enzyme the limiting factor halting the production. Figure 2. looks at the pH change in distilled water added to the catechol oxidase and potato juice extract. By the data shown very acidic and very basic solutions were unable to produce as much benzoquinone. Bringing the enzyme to a more neutral state speeds up the production. The standard curve shows this by the peak at the concentration of pH 7.
Looking at the solutions placed in different temperatures the data in figure 3 shows most production at 20-degrees F and 35-degrees F. Being at these temperatures are closest to room temperature, which is 25-degreesF, the enzyme produced best stored at this temperature. Discussion Production Of Benzoquinone With Graph Data Through the group results, seen in figure1, figure 2, and figure 3, the mean brought very similar absorption readings. The catechol oxidase with potato juice extract seemed to vary in production in pH data collected, though there was still gradual increase in production.
Temperature solutions showed common production the 70- degrees F temperature and 3-degrees F temperature. As for the number of drops of catechol oxidase added to the solution proved that more of the enzyme present will lead to greater production of benzoquinone. Common Errors and Ways to Improve The amounts of drops that were put into the test tubes may not have been exactly the correct amount. This was exemplified through the test tube labeled 70 degrees. The absorption value was 90, most likely because more drops of potato juice extract or catechol were added to the test tube then needed.
These unexpected results could be tested by redoing the procedure and being critical about how many drops each test tube was given. Production of Benzoquinone The results of the three procedures; substrate amount, pH, and temperature were fairly close to the hypothesis. The hypothesis was when there is a greater presence of catechol; a greater amount of benzoquinone will be produced up until the present amount of catechol oxidase becomes limited due to its participation in the reactions. All three of the procedures supported this hypothesis. The reliability of the results that were obtained is slightly lower than what would be wanted.