Biologically Important Molecules: Enzymes
Introduction
The purpose of this experiment is to determine how outside factors—pH, temperature, and incubation time—affect an enzyme’s ability to catalyze, or speed up, a chemical reaction and generate the maximum amount of product.
The incubation medium’s pH level, for example, may directly influence the binding of the substrate to the enzyme’s active site. This results from the correlation between pH and the ionization of the substrate and amino acid side chains at the binding site of the enzyme. Extreme pH levels may also distort, or even denature, the enzyme (Bender, 2016).
Temperature is another factor that may affect the amount of product. Heat provides energy for the reaction, thereby increasing
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The longer an enzyme incubates with its substrate, the more product is formed. However, if the incubation is too long, the enzyme will denature, thus decreasing the amount of product (Bender, 2016).
Therefore, I hypothesize that if an enzyme is to function at its highest capacity, then it must be found in its optimal levels of pH, temperature, and incubation time. This experiment will examine five different enzymes to try to determine these optimal conditions needed to generate the most product. It will test each independent variable—pH, temperature, and incubation time—for all five enzymes to determine the most favorable conditions for each to produce the greatest amount of catalytic
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Under these conditions, the enzyme generated 93.70 μmol of product.
Figure 5
Optimal Incubation Conditions for Enzyme 3 Figure 6
Table of Optimal Values for Enzyme 3 The optimal conditions for Enzyme 4 (shown by the tables in Figure 7 and Figure 8) are a pH range of 8.80 to 9.30, with a peak at 9.050; a temperature range of 30.0°C to 50.0°C, with a peak at 42.0°C; and an incubation time range of 50.0 minutes to 60.0 minutes, with a peak at 59.0 minutes. Under these conditions, the enzyme generated 101.090 μmol of product.
Figure 7
Optimal Incubation Conditions for Enzyme 4 Figure 8
Table of Optimal Values for Enzyme 4 The optimal conditions for Enzyme 5 (shown by the tables in Figure 9 and Figure 10) are a pH range of 2.20 to 2.80, with a peak at 2.680; a temperature range of 30.0°C to 50.0°C, with a peak at 38.0°C; and an incubation time range of 50.0 minutes to 60.0 minutes, with a peak at 60.0 minutes. Under these conditions, the enzyme generated 99.880 μmol of
As previously stated, temperature is another variable that can affect the enzyme reaction rate. At low temperatures there will be little to no activity but as the temperature is increased so the rate of activity will increase. However, just like pH, once the optimal temperature is reached and surpassed, the enzyme will begin to break apart. The enzyme will be experimentally tested using temperatures around room temperature (27℃). Due to the fact that there is a human application the enzyme will continue to be tested until temperatures of human death (45℃). After the testing is completed it can be deduced what temperatures create an optimum environment for the normal functioning of the enzyme. One
Background and Introduction: Enzymes are proteins that process substrates, which is the chemical molecule that enzymes work on to make products. Enzyme purpose is to increase the rate of activity and speed up chemical reaction in a form of biological catalysts. The enzymes specialize in lowering the activation energy to start the process. Enzymes are very specific in their process, each substrate is designed to fit with a specific substrate and the enzyme and substrate link at the active site. The binding of a substrate to the active site of an enzyme is a very specific interaction. Active sites are clefts or grooves on the surface of an enzyme, usually composed of amino acids from different parts of the polypeptide chain that are brought together in the tertiary structure of the folded protein. Substrates initially bind to the active site by noncovalent interactions, including hydrogen bonds, ionic bonds, and hydrophobic interactions. Once a substrate is bound to the active site of an enzyme, multiple mechanisms can accelerate its conversion to the product of the reaction. But sometimes, these enzymes fail or succeed to increase the rate of action because of various factors that limit the action. These factors can be known as temperature, acidity levels (pH), enzyme and/or substrate concentration, etc. In this experiment, it will be tested how much of an effect
Using an O2 sensor to measure how much additional oxygen is added into a sealed Nalgene bottle, we could determine the enzymes effectiveness. Three variations of temperature were tested: cold, hot and room temperature. This was done by taking test tubes with the three different samples and placing them in a Nalgene Bottle. It was determined that the enzyme is most effective when at room temperature although still functions when it is chilled. The heated sample was drastically less effective.
These results show how temperature of extreme high, or low affects enzyme activity. The highest rate of enzyme activity occurred at 37 Cº. Anything that was hotter or cold than 37 Cº slowed the reaction rate. As I thought, 100 degrees would denature the enzyme, and that was the case. The data provided shows exactly what temperatures enzymes work best, and worst. The objective was achieved as we discovered the different reaction rates under different temperatures. The results are reliable, as we know enzymes do not work well when under extreme heat or denaturation occurs. What I learned in this experiment was that enzymes don’t work well under cold temperatures because they tend to move slower. My hypothesis did not quite match, because I thought they work best at lower temperatures.
As stated in the introduction, three conditions that may affect enzyme activity are salinity, temperature, and pH. In experiment two, we explored how temperature can affect enzymatic activity. Since most enzymes function best at their optimum temperature or room temperature, it was expected that the best reaction is in this environment. The higher the temperature that faster the reaction unless the enzyme is denatured because it is too hot. Similarly, pH and salinity can affect enzyme activity.
Introduction: Enzymes are protein catalysts facilitating the conversion of substrates into products (Alexander and Peters, 2011). They go through a whole chemical reaction which starts off with the substrate and then ends up with a product. The only way this reaction can be adjusted or not even work is if they end up going through some sort of affect which only temperature and pH levels can do determining the environment. When enzymes are in an environment that is too acidic or alkaline, their chemical properties, sizes and shapes can become altered (Magher, 2015) Chemical modification of proteins is widely used as a too; to maintain a native conformation, improving stability (Rodriguez-Cabrera, Regalado, and Garcia-Almendarez, 2011) In this experiment, four trials were conducted and recorded every 15 seconds for 5 minutes in order to calculate the optimum levels and IRV.
pH along with temperature and concentration is a limiting factor of enzyme activity. Enzymes, which are proteins, have a distinct pH range in which they work most effectively. Beyond or below this specific pH, enzyme activity begins to diminish and eventually the enzyme is rendered useless. The optimum pH of an enzyme is the pH at which it is most effective and causes the fastest rate of reaction(source). This decrease in enzyme activity beyond the optimum pH is attributed to the change in shape of the active site(explained earlier) of the enzyme. This change in shape can be brought upon when the surrounding pH moves away from the optimum pH, and it can prevent the enzyme from binding with the substrate.
The motive of this lab is to attain a better understanding of enzyme activity by timing chemical reactions in certain temperatures and pH levels. Enzymes act as catalysts that help speed up reactions. Without these enzymes chemical reactions in metabolism would be backed up. There are two factors that affect an enzyme’s reaction rate: temperature and pH levels. In this label we will be testing different pH levels and temperatures to see which ones cause the most reactions.
Corporation, W. B. (2013). Introduction to Enzymes. Retrieved 2013 йил 27-2 from Worthington Biochemical Corporation: https://gateway.emmanuel.qld.edu.au/cvpn/aHR0cDovL3d3dy53b3J0aGluZ3Rvbi1iaW9jaGVtLmNvbQ/introbiochem/effectsph.html
Enzymes have an ideal range of values for any of the variables, or optimal conditions, in this experiment. When these optimal conditions are
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
The independent variable in this investigation is pH. Each individual enzyme has it’s own pH characteristic. This is because the hydrogen and ionic bonds between –NH2 and –COOH groups of the polypeptides that make up the enzyme, fix the exact arrangement of the active site of an enzyme. It is crucial to be aware of how even small changes in the
The purpose of this experiment was to record catalase enzyme activity with different temperatures and substrate concentrations. It was hypothesized that, until all active sites were bound, as the substrate concentration increased, the reaction rate would increase. The first experiment consisted of five different substrate concentrations, 0.8%, 0.4%, 0.2%, 0.1%, and 0% H2O2. The second experiment was completed using 0.8% substrate concentration and four different temperatures of enzymes ranging from cold to boiled. It was hypothesized that as the temperature increased, the reaction rate would increase. This would occur until the enzyme was denatured. The results from the two experiments show that the more substrate concentration,
Enzyme catalysis is dependant upon factors such as concentration of enzyme and substrate, temperature and pH. These factors determine the rate of reaction, and an increase in temperature or pH above the optimum will
To study the effects of temperature, pH, enzyme concentration, and substrate concentration there were certain steps that were followed in order to conduct this experiment. Each factor had a separate procedure to follow to find how each had a different effect on the enzyme.