Introduction: So what is an enzyme? An enzyme is “any protein that acts as a catalyst, increasing the rate at which a chemical reaction occurs. The human body probably contains about 10,000 different enzymes.”(Farlex) The active site or sites of every enzyme are composed of a particular array of amino acids. The active site exhibits specificity for the substrate of the enzyme. The enzyme should bind and catalyze the reaction for its specific substrates better than with any other substrates. Therefore, the following experiment is testing the ability of lactase to specifically bind and interact with lactose compared to maltose. Materials and Methods: Enzyme Specificity To begin with the experiment, two microfuge tubes were labeled. …show more content…
The EDTA tube required 0.5 M EDTA to be added until the solution reached 1.25 mL of EDTA. To the Control tube, distilled water had to be added. Once done, this tube now contained 1.25 mL of water. After these steps were done, each tube called for three drops of milk. The tubes must then be inverted and allowed to sit for one minute. Once one minute had gone by, each tube needed three drops of lactase solution. For now, both tubes had to be placed in the 40 °C water bath and left alone for ten minutes. After the ten minute timer went off, the two tubes were taken out of the water bath. A glucose strip had to be placed in each tube for one second each. Once that had been done, both glucose strips had to be set out on the lab bench for thirty seconds. At the end of thirty seconds, the strips and their new coloration had to be compared to the chart provided, which helps to determine the amount of glucose in mg/dL. Results: 8.3 The results for enzyme specificity were determined by the amount of glucose present in the lactose and maltose microfuge tubes. Inside the lactose labeled tube, the average glucose measurement was 1,700 mg/dl. (Table 1) On the other hand, the tube labeled Maltose had an average glucose measurement of 20 mg/dl. (Table 1) Substrate Lactose Maltose Glucose (mg/dL) 1,700 mg/dl 20
Data from Table 1. confirms the theory that as the concentration of glucose increases so will the absorbance of the solution when examined with the glucose oxidase/horseradish peroxidase assay. Glucose within the context of this assay is determined by the amount of ferricyanide, determined by absornace, which is produced in a one to one ratio.1 Furthermore when examining the glucose standards, a linear calibration curve was able to be produced (shown as Figure 1). Noted the R2 value of the y = 1.808x - 0.0125 trend line is 0.9958, which is statistically considered linear. From this calibration curve the absorbance values of unknowns samples can be compared, and the correlated glucose concentration can then be approximated.
We then took 1ml of the 0.1% solution from test tube 2 using the glucose pipette and added it to test tube 3, we then used the H2O pipette and added 9ml of H2O into test tube 3 creating 10ml of 0.01% solution.
3.) Divide your 30g of white substance into the 4 test tubes evenly. You should put 7.5g into each test tube along with the water.
These labels indicated the lactose solution that was be placed into the mini-microfuge tubes. The varying lactose ph solutions were obtained. The four miniature pipets were then used, (one per solution,) to add 1mL of the solution to the corresponding mini-microfuge tubes. When this step is completed there were two mini-microfuge tubes that matched the paper towel. Then, once all of the solutions contained their respective lactose solutions, 0.5mL of the lactase enzyme suspension was added to the first mini-microfuge tube labeled LPH4 on the paper towel, and 4 on the microfuge tube. As soon as the lactase enzyme suspension was added to the mini-microfuge tube, the timer was started in stopwatch mode (increasing.) When the timer reached 7 minutes and 30 seconds, the glucose test strip was dipped into the created solution in the mini-microfuge tube for 2 seconds (keep timer going, as the timer is also needed for the glucose strip. Once the two seconds had elapsed, the test strip was immediately removed, and the excess solution was wiped gently on the side of the mini-microfuge tube. The timer was continued for 30 addition seconds. Once the timer reached 7:32 (the extra two seconds accounting for the glucose dip), the test strip was then compared the glucose test strip color chart that is found on the side of the glucose test strip
The shape of the molecules is changing and so the enzyme molecules can no longer fit into the gaps in the substrate that they need to and therefore the enzymes have de – natured and can no longer function as they are supposed to and cannot do their job correctly. Changing the temperature: Five different temperatures could be investigated. Water baths were used to maintain a constant temperature. Water baths were set up at 40 degrees, 60 degrees and 80 degrees (Celsius). Room temperature investigations were also carried out (20 degrees).
The purpose of this experiment was to discover the specificity of the enzyme lactase to a spec...
Purpose: The purpose of this lab is to explore the different factors which effect enzyme activity and the rates of reaction, such as particle size and temperature.
6. Place the test tube in the beaker. Secure the test tube and thermometer to the retort stand using clamps. Begin heating the water bath gently.
The three-dimensional contour limits the number of substrates that can possibly react to only those substrates that can specifically fit the enzyme surface. Enzymes have an active site, which is the specific indent caused by the amino acid on the surface that fold inwards. The active site only allows a substrate of the exact unique shape to fit; this is where the substance combines to form an enzyme- substrate complex. Forming an enzyme-substrate complex makes it possible for substrate molecules to combine to form a product. In this experiment, the product is maltose.
...remain the same at 4ºC and 25ºC. The final result of this experiment was that glucose was more present in environments of higher temperatures. Our hypothesis and predictions were wrong because lower temperatures do not break down the enzymes because they become denatured. The enzyme activity decreases once the temperature decreases, as well. Enzyme activity increases when there is a rise in temperature, which is why lactose is broken down in much higher temperatures, resulting in a high presence of glucose.
2. In the large beaker, put water and boil it completely. After that, remove the beaker from heat. 3. Sample tubes (A-D) should be labeled and capped tightly.
In this experiment, researchers used different measurements of catechol and 1cm of potato extract. Researchers hypothesized that the increase in substrate would level out the enzyme activity by
tube. Add 6 mL of 0.1M HCl to the first test tube, then 0.1M KMnO4 and
Place the dialysis tubing into the cup it corresponds with 11. Fill the first cup with the solution indicated on the outside of it until the solution completely covers the model cell placed inside. 12. After 24 hours remove the dialysis tubing bag from the cup. Once again gently roll it back and forth on a paper towel to remove excess liquid.
Introduction Most mammals are initially born with the ability to break down the polysaccharide, lactose into smaller monosaccharides but at an early age, usually as the child starts to rely less on their mother for direct nurturance, this ability ceases. This inability to break down lactose is known as lactase non-persistence. Lactase Non-persistence is the wild type in the population surprisingly even though a vast majority of the population is lactase non persistent. Individuals with the ability to digest the lactose found in milk are considered lactase persistent or lactose tolerant.