Effect of Varying Temperatures on Enzymatic Activity of Bacterial and Fungal Amylase and Hydrolysis of Starch

The Effects of Varied Temperatures, pH Values, Enzyme Concentrations, and Substrate Concentrations on the Enzymatic Activity of Catecholase

Amylase experiment # 2 was done to see how the pH affected the efficacy of the enzyme. First we collected all of the materials that were necessary to make this experiment. We needed five clean test tubes, the following standard solutions, 1% Starch Solution pH 3,1% Starch Solution pH 5,1% Starch Solution pH 7,1% Starch Solution pH 9,1% Starch Solution pH 11

Temperature controls the speed the enzymes work at. Higher temperatures increase the kinetic energy which increases the chance of collision therefore speeding up the rate of

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.

The purpose of this experiment was to determine (1) the reaction rate of an amylase enzyme in starch and (2) the environmental factors that can affect the enzymatic activity. The hypothesis, in relation to the enzymatic activity by variables such as the substrate concentrations, temperature, PH and chemical interactions on the rate of reaction, stated

Hydrolysis of starch for fungal amylase Aspergillus Oryzae and bacterial amylase Bacillus Licheniformis at different temperatures.

2. What effect did boiling and freezing have on enzyme activity? Why? How well did the results compare with your prediction?__Boiling caused amylase to be denatured, thus inactivating the enzyme. Freezing has no effect. The function of an enzyme is directly related to its environment, like temperature.__

During these experimental procedures, the implication of multiple different temperatures on fungal and bacterial amylase was studied. In order to conduct this experiment, there were four different temperatures used. The four temperatures used were the following: 0 degrees Celsius, 25 degrees Celsius, 55 degrees Celsius, and 80 degrees Celsius - Each temperature for one fungal and one bacterial amylase. Drops of iodine were then placed in order to measure the effectiveness of the enzyme. This method is produced as the starch test. The enzyme was tested over the course of ten minutes to determine if starch hydrolysis stemmed. An effective enzyme would indicate a color variation between blue/black to a more yellowish color towards the end of the time intervals, whereas a not so effective enzyme would produce little to no change in color variation. According to the experiment, both the fungal amylase and bacterial amylase exhibited a optimal temperature. This was discovered by observing during which temperature and time period produced a yellow-like color the quickest. Amylase shared a similar optimal temperature of 55 degrees Celsius. Most of the amylases underwent changes at different points, but some enzymes displayed no effectiveness at all. Both amylases displayed this inactivity at 0 degrees Celsius. At 80 Celsius both the enzymes became denatured due to the high temperatures. In culmination, both fungal and bacterial amylase presented a array of change during it’s

test the pH of the amylase a drop of the solution should be put on pH

Amylase is an enzyme that is located in human saliva. It is solely accountable for breaking down starch as a way to start the breakdown of food and is one of the first steps of digestion. The time at which the enzyme starts the chemical reaction with starch is called the reaction rate. In order to study how amylase works against starch, this experiment consisted of two tests; each testing a different condition of amylase. The first test was to simply study the reaction between saliva and amylase and note the reaction rates. The second test was to see if increasing the pH would decrease the reaction rate or halt it all together. Saliva was collected, diluted, and tested for reactions between starch and amylase. Another sample of saliva was collected, diluted, and had its pH increased and tested for reaction rate. The findings after the experiment was conducted aligned with the original hypothesis. The change in pH did show a significant decrease in the reaction rate.

Record observations: what color did the solution turn? Orange or blue/ black? This will tell whether the amylase is able to denature the starch solution Orange=Negative Blue/black= Positive

In this experiment we wanted to determine the optimal temperatures for fungal, Aspergillus oryzae, and bacterial, Bacillus licheniformis. In order to see if any of the starch was broken down, Iodine was mixed with the starch-amylase substance. In four spot plates, the groups labeled the different temperatures, once the iodine came in contact with the starch, the result would be a reaction that turns the fluid into this dark blue/black color. In a span of 10 minutes, with occasional check ups on the solution every 2 minutes, the amylase-starch solution was placed into five types of temperature, all being Celsius. The five temperatures were 0 degrees, 25 degrees, 55 degrees, and 85 degrees Celsius. The solution would change colors, so in order to measure the changes, a scale was used. Such scale was a 1-5 scale, with colors next to each number. One being the lightest color, or yellow, and 5 being the darkest color, or black. Based on the change of color, we could tell how fast it hydrolyzed the starch in a span of 10 minutes. To keep record of the results, the results were put in Data Tables used from the Lab Manual. The average optimal temperature for Bacteria Amylase was 85 degrees Celsius, while the Fungal was 55 degrees Celsius. You can see this by looking for the

The amount of the reducing sugars like glucose or inverted sugar contained in the sample is calculated from the quantity of the Cu2+ from the Fehling solution which have undergone a reaction with the sugar, by using the Bertrand tables [4]. RESULTS AND DISCUSSION To study the optimal conditions for the hydrolysis of starch with the a – amylase enzyme XK – 60 under consideration, we have varied the following factors: concentration of the substrate, concentration of the enzyme, temperature of the hydrolysis, pH and the influence of inhibitors. In order to determine the optimal concentration of the substrate we have done experiments in the range from 100 to 300 g.l-1 starch. From the results presented in Table 1 it is evident that for substrate concentrations 250 and 300 g.l-1, the level of hydrolysis is almost the same (30.1 % and 30.3 % respectively). We supposed that this was most likely due to the inhibiting action of the reducing sugars obtained during hydrolysis at substrate concentrations higher than 250 g.l-1. To prove this hypothesis, we investigated the influence of the concentration of the glucose added to the substrate on the rate of the enzymatic reaction. As can be seen from Fig. 1, the addition of glucose in the beginning of the hydrolysis process significantly reduces the reaction rate of the process. The

This lab is on the productivity of bacterial and salivary amylase in different pH levels and temperature. To measure the productivity of the amylase, we will add starch to amylase samples (gives the amylase a job) and Lugol’s iodine (to see if it is properly doing its job by converting that starch into sugar. Light/transparent color means little to no starch, and a dark color means an abundance of starch). The pH will be controlled by adding Hydrochloric Acid (HCl-acidic) or Sodium hydroxide (NaOH-basic). Temperature will be controlled by first boiling starch-amylase test tubes then placing 2 in a cold bath, leaving 2 at room temperature, and putting 2 in a hot bath.

Throughout history, there have been many studies conducted regarding different biological branches of the complex study of life. Amylase, an enzyme that catabolizes starch polymers, is one of the most important enzymes needed for the production of certain foods, such as syrups, and different processes such as fermentation. Like everything’s biological nature, these certain enzymes are affected by different factors ranging from pH levels to temperature. Finding out the temperature at which these enzymes reach their optimal condition (conditions at which these enzymes work the best), is one of the most revealing studies of all. In this experiment, the affect that temperature has on the enzymatic activity of Amylase was the leading role, whether it was to determine if it was slowing it down or speeding it up. For both experiments, iodine was used as the control. First, the optimal temperatures of fungal Amylase were tested. For this particular task, there was not too much changed noticed. For most of the temperatures, the activity change seen was very minimal. With iodine and starch by itself, the activity remained the same as well as with the amylase at the 85 degrees test. When testing for the bacterial Amylase, the most change was seen at the 55 degrees test. Overall, it can be said that as temperature rises or decreases, enzymatic activity is affected, but of course with a certain plateau.