This experiment is to study and measure the enzyme activity of β-galactosidase in the different concentrations of o-Nitrophenylgalactoside (ONPG) using a spectrophotometer. The spectrophotometer was also set at 420nm, a wavelength which is best for recording the absorbance values for the experiment. From the results, 0.9mM ONPG solution has the highest absorbance and 0.1mM ONPG solution has the least. Also, 0.5mM ONPG solution has the highest rate of enzyme activity and it is the most efficient as the enzyme activity of the ONPG solution continues even though the other concentrations of ONPG solution has already stopped the enzymatic reactions as the substrate is already used up.
Of the many functions of proteins, catalysis is by far the most vital. When catalysis is not present, most reactions in the biological systems take place very slowly to produce at an adequate pace for metabolising organism. The catalysts that take this role are called enzymes. Enzymes are the most efficient catalysts; they can enhance rate of reaction by up to 1020 over uncatalysed reactions. (Campbell et al, 2012).
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
Lab six requires students to observe the effects of pH and enzyme concentration on catecholase activity. Enzymes are organic catalysts that can affect the rate of a chemical reaction depending on the pH level and the concentration of the enzyme. As pH comes closer to a neutral pH the enzyme is at its greatest effectiveness. Also at the absorbance of a slope of 0.0122 the enzyme is affected greatly. The pH effect on enzymes can be tested by trying each pH level with a pH buffer of the same pH as labeled as the test tube and 1mL of potato juice, water, and catechol. This is all mixed together and put in the spectrophotometer to test how much is being absorbed at 420nm. As the effect on enzyme concentration can be tested almost the same way. This part of the exercise uses different amounts of pH 7-phosphate buffer and potato juice, and 1mL of catechol mixed together in a test tube. Each substance is put in the spectrophotometer at a wavelength set tot 420nm. The results are put down for every minute up to six minutes to see how enzyme concentration affects reaction rate. The results show that the pH 8 (0.494) affects the enzyme more than a pH of 4 (0.249), 6 (0.371), 7 (0.456), and 10 (0.126). Also the absorbance is greatest at a slope of 0.0122 with test tube C that has more effect on the reaction rate, than test tube A, B, and D.
These results shown from this experiment led us to conclude that enzymes work best at certain pH rates. For this particular enzyme, pH 7 worked best. When compared to high levels of pH, the lower levels worked better. The wrong level of pH can denature enzymes; therefore finding the right level is essential. The independent variable was the amount of pH, and the dependent being the rate of oxygen. The results are reliable as they are reinforced by the fact that enzymes typically work best at neutral pH
Enzymes are an organic substance that are made up of polymers of amino acids that help the digestive system and metabolic processes in living organisms (Funk and Wagnalls, 2016). Enzymes are able
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,
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
Enzymes are a key aspect in our everyday life and are a key to sustaining life. They are biological catalysts that help speed up the rate of reactions. They do this by lowering the activation energy of chemical reactions (Biology Department, 2011).
Introduction:Enzymes are made up of proteins which are produced within living cells and act as catalysts which speed up chemical reactions. They are made up of long chains of amino acids containing carbon, hydrogen, oxygen and nitrogen. Enzymes are structured to be
To begin the lab, part A was performed to determine the amount of enzyme that would produce a reaction rate that did not proceed too slow or too rapidly. As seen on page 13 of the Lab Handout, varying amounts of tyrosinase and phosphate buffer were added to a cuvette while the amount of the L-DOPA was constant. After all reagents were added to the cuvette, the cuvette was inserted into spectrophotometer and absorbance of product formation at 475 nm was recorded for two minutes at fifteen seconds interval. After absorbance of product formation was measured and recorded for each cuvette, as shown on page 14 of the Lab Handout, graphs of rate of production formation versus time, were made with the data of each cuvettes and the
Enzyme is the fastest known catalyst. They increase the velocity and rate of reactions in living cells. Diverse range of proteins works as enzyme with their specific temperature point, pH etc. Enzymes react with the molecules known as substrates. Binding site of an enzyme and a substrate is known as active site. The amount of enzyme present in a reaction is measured by the activity it catalyzes. The relationship between activity and concentration is affected by many factors such as temperature, pH, etc. Though enzymes are important for biological reactions, certain factors play an important role in the
Enzymes are proteins that are essential to our body because it acts as a catalase to speed up chemical reactions within the body using less energy. Enzyme is very specific, it works like a key and lock method by having the exact substrate binding to the enzyme’s active site. Even though enzyme can be reuse to a point that they are denatured. When something is denatured there shape become deform and no longer works. When enzyme doesn’t work our body reactions slows down to a point where it all stops together. All Enzymes reaches there optimal level before it slows down, enzyme also have a specific range of pH and temperature that they can function through. For instance the enzyme that is presented in the mouth can endure a pH around 7 while those in the stomach can endure a pH of 2.0 if the enzyme were to go to the wrong pace than they would become denatured because they can handle the accidences or the bases produced by the organs. Enzyme can also be denatured if they are exposing to extreme heat. As the temperature gets colder the rate of the enzyme will become less efficient and when it’s hotter the heat generate more kinetic energy for the enzyme and substrate making
What is really interesting and important to know about enzymes is the fact that they can continuously cycle, which means that they can go through various chemical reactions and remain unchanged. Although enzymes can do all of these important things in our body, they can still be destroyed or better yet “denatured” since they are proteins. Denaturation is a process where a protein loses its structure and function. This can be caused by increases in temperature and changes pH levels (Vodopich and Moore 2).