Atpase Lab Report

In this lab or experiment, the aim was to determine the following factors of enzymes: (1) the effects of enzymes concentration the catalytic rate or the rate of the reaction, (2) the effects of pH on a particular enzyme, an enzyme known and referred throughout this experiment as ALP (alkaline phosphate enzyme) and lastly (3) the effects of various temperatures on the reaction or catalytic rate. Throughout the experiment 8 separate cuvettes and tubes are mixed with various solutions (labeled as tables 1,3 & 4 in the apparatus/materials sections of the lab) and tested for the effects of the factors mentioned above (concentration, pH and temperature). The tubes labeled 1-4 are tested for pH with pH paper and by spectrophotometer, cuvettes 1a-4a was tested for concentration and cuvettes labeled 1b-4b was tested for temperature in four different atmospheric conditions (4ºC, 23ºC, 32ºC and 60ºC) to see how the enzyme solution was affected by the various conditions. After carrying out the procedures the results showed that the experiment followed the theory for the most part, which is that all the factors work best at its optimum level. So, the optimum pH that the enzymes reacted at was a pH of 7 (neutral), the optimum temperature that the reactions occurs with the enzymes is a temperature of 4ºC or

In the experiment we used Turnip, Hydrogen Peroxide, Distilled Water, and Guaiacol as my substances. On the first activity, Effect of Enzyme concentration of Reaction Rate for low enzyme concentration, we tested three concentrations of the turnip extract, and hydrogen peroxide. For the Turnip Extract I used 0.5 ml, 1.0 ml, and 2.0 ml. For hydrogen peroxide we used 0.1 ml, 0.2 ml, and 0.4 ml. We used a control to see the standard, and used a control for each enzyme concentration used. The control contains turnip extract and the color reagent, Guaiacol. We prepared my substrate tubes separately from the enzyme tubes. My substrate tube

From Figure 5, Vmax = 0.0172 and Km = 0.3144. At ½ Vmax (Figure 4), the rate of reaction increases steadily with an increase in substrate concentration. At this point, the rate of product formation is partially limited by substrate availability. As the curve approaches Vmax, the enzyme catalyzed saturation increases. In effect, the enzyme activity increases the rate of product formation up to a point whereby the reaction rate remains constant as more substrate is added (saturation).

Gammarus setosus is a marine amphipod that is found in the intertidal waters of British Columbia. A study of temperature on the rate of movement of Gammarus setosus was undertaken to find whether temperature would positively or negatively affect the activity of Gammarus setosus. Specimens of Gammarus setosus was obtained and tested. 30 ppt salt water of 5°C, 23°C, and 30°C was used to perform the experiment. The specimens were allowed to acclimate for one minute. Then, the distance traveled by the specimens was collected and analyzed. A total of 13-15 replicates were tested per trial. It was found that the specimens were most active in the 20°C trial. The

Enzymes are catalysts that function to speed up reactions; for example, the enzyme sucrose speeds up the hydrolysis of sucrose, which breaks down into glucose and fructose. They speed up reactions but are not consumed by the reaction that is taking place. The most important of the enzyme is the shape as it determines which type of reaction the enzyme speeds up. Enzymes work by passing/lowering and energy barrier and in doing so; they need to bind to substrates via the active. Once they do, the reaction speeds up so much more quickly than it would without the enzyme. Coenzymes and cofactors aid the enzyme when it comes to binding with the substrate. They change the shape of the active site so the substrate can bind properly and perform its function.

2. We measured 1 mL of turnip peroxidase (the enzyme) and 3 mL of neutral buffer (pH corresponding to the test tube number i.e. pH 5 in test tube 5) with a syringe and disposed it into tubes 3, 5, 6, 7, 8, and 10

350 is the maximal substrate concentration. The max initial reaction rate cannot be reached at low lactose concentrations because there would need to be more substrate added.

The hypothesis tested in this experiment was, if the temperature of enzyme catalysis were increased, then the reaction rate would increase, because enzyme-catalysis reacts by randomly colliding with substrate molecules, and the increase in temperature increases the speed of collision or reaction rate. The final data collected for the experiment was positive with my hypothesis. The coffee filter, covered in potato solution, sank and rose at a faster pace in the hydrogen peroxide when the temperatures were raised.

Lactose is a sugar that can be put into smaller molecules, glucose and galactose. Lactose is when you are not able to digest milk and dairy meaning that the enzyme lactase that breaks down lactose is not functioning properly. ONPG was used as a substitute for lactase because even though it is colorless it helps show enzyme activity by turning yellow. This experiment measured the absorbance ONPG when exposed to lactase within an environment of different salinity’s. The enzyme, lactase, was obtained by crushing a lactaid pill and then was added into four cuvettes. ONPG and salt solution of different concentrations were added and their levels of absorption was measured by a spectrophotometer. The results showed that higher salt concentrations have a lower level of absorption. There were 4 cuvettes and within those cuvettes that solutions within them were being tested and the results showed the more salt solution added with the lactase the lower the absorbance. The less salt solution there was a higher rate of absorbance. The data supported the hypothesis that with increasing NaCl concentration there would be a decrease in enzyme activity.

J. Moldovan & B. Nilson, (2010), Lab 4 – Enzyme Kinetics, UBCO BIOL/BIOC 393, UBC Vista accessed Monday, November 8th, 2010.

Morgan Calafati #1 MAKE OBSERVATIONS Even in middle school biology I learned that the body produced ATP and used it for energy, and I never thought about what the abbreviation meant. When I learned that ATP was adenosine triphosphate, which is composed of a modified form of adenine and phosphates, it made me wonder why there is not chemical energy using the other bases. #2 ASK QUESTIONS Why is ATP the most common form of chemical energy rather than CTP, TTP, or GTP?

Describe an experiment (assay) that could be used to determine the kinetic parameters of your enzyme.

Organisms cannot depend solely on spontaneous reactions for the production of materials because they occur slowly and are not responsive to the organism's needs (Martineau, Dean, et al, Laboratory Manual, 43). In order to speed up the reaction process, cells use enzymes as biological catalysts. Enzymes are able to speed up the reaction through lowering activation energy. Additionally, enzymes facilitate reactions without being consumed (manual,43). Each enzyme acts on a specific molecule or set of molecules referred to as the enzyme's substrate and the results of this reaction are called products (manual 43). As a result, enzymes promote a reaction so that substrates are converted into products on a faster pace (manual 43). Most enzymes are proteins whose structure is determined by its sequence of its amino acids. Enzymes are designed to function the best under physiological conditions of PH and temperature. Any change of these variables that change the conformation of the enzyme will destroy or enhance enzyme activity(manual, 43).

The process that generates the most ATPs is fatty acid oxidation. This is because β-oxidation of palmitate which is a common saturated fatty acids in most living organisms which could produce 106 net yield of ATPs.palmitate with its 16 carbon give below overall reaction: C 16:0-CoA + 7 FAD + 7 NAD+ + 7 H20 + 7 CoA → 8 acetyl-CoA + 7 FADH2 + 7 NADH + 7 H+ 2.5 ATPs per NADH, 1.5 ATPs per FADH2 and 10 ATPs per acetyl CoA produced while 2 ATPs consumed during the activation of palmitate to acyl-CoA gives net yield of 106 ATPs. As for citric acid cycle, oxidation of 1 acetyl-CoA can give a total of 10 ATPs and it is based on the below equation: Acetyl-CoA + 3 NAD+ + Q + GDP + Pi +2 H20

The hypothesis is as the substrate concentration has an increase so will the reaction of velocity if the amount of enzyme is kept constant.