Fungal Amylase Lab Report

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

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 optimal temperature of Bacillus lichenformis bacterial amylase and Aspergillus oryzae fungal is determined by mixing a starch solution into the bacterial and fungal amylases that are put in four different temperatures (0, 20, 55, 85 degrees Celsius). Then after every two-minutes, ending at the ten-minute mark, a small sample of the starch-amylase mixture is put into a well with a couple drops of iodine to help show the change in starch. This was done because when iodine is exposed to starch it changes color. Based on the color chart given in our lab manuals, the reaction of the amylase to the starch solution will give the starch-amylase mixture in the iodine a yellow color to signify if the presence of solely iodine and/or little starch depending on temperature. This means that the amylase broke down the starch solution because its temperature was optimal. Majority of the results came out black or dark brown therefore the amylase wasn’t put in the proper temperature to break down the starch solution at a faster pace. The temperature that seemed most optimal was at 55 degrees Celsius for both fungal and bacterial because it showed a more brown to yellowish color when put into the iodine. That showed that the amylase was able to break down the starch at a faster rate because it was working at its optimal temperature.

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

An experiment was performed to test how temperature variations affect enzymatic activity of the enzyme amylase. The results of the experiment will also determine the optimal temperature of the amylase enzyme. The results of the experiment provide evidence for determining the environments that the enzyme amylase would most likely be present. By determining the possible environments, one can predict what and how environmental factors will affect the enzyme amylase. Two forms of amylase (Bacterial - Bacillus licheniformis and Fungal - Aspergyllus oryzae) were combined with starch molecules at four different temperatures (0⁰, 25⁰, 65⁰, 85⁰ Celsius). The combination of starch and the amylase enzyme resulted in a visual chemical reaction that was recorded. The enzyme activity was recorded every two minutes, starting at 0 and ending at 10. The start time 0 served as the control group of the experiment. The results concluded that both bacterial and fungal amylase has an optimal temperature around 65⁰C. This was possible to determine by recording the color change of the spot plate wells. Amylase catalyzes efficiently at its optimal temperature which resulted in yellow spot plate wells. Enzymatic activity decreased when the temperature was less than 65⁰C, resulting in a green-brown well. The green wells indicated that starch wasn’t broken down completely and was still present. Temperatures greater than 65⁰C resulted dark-green wells which resembled the denaturing of

An enzyme also known as a protein, is a biological catalyst which speeds up chemical reactions by lowering the activation energy to increase the rate in which the reaction occurs. The enzyme used was amylase, which breaks down starch molecules into maltose. PH, substrate concentration, salt concentration, and temperature. When enzymes reach a low temperature, the activity is slowed down of molecule movement, but the enzyme is not destroyed. Once enzymes are placed in optimal temperatures once again, it will restore its activity to a normal rate. When enzymes reach too high above optimal temperature, the enzyme is denatured and cannot be restored. In the experiment performed the activity of breaking down starch in fungal and bacterial amylase was being tested at a range of temperatures and time. The fungal and bacterial amylase work best at optimal temperature. Amylase will function best at sixty degrees Celsius at 10 minutes when starch had been one hundred percent hydrolyzed. Hydrolyzed is the breakdown of molecules through addition of water. The experiments independent variables were the time, temperature and enzyme used. The dependent variable was the enzyme activity that broke down the starch into maltose. The controlled variables were the temperature baths, the iodine drop amount, the mixture drop amount, and location of experiment. The control group was the zero minutes without amylase at

amylase enzyme and the optimal temperature for fungal and bacterial amylase. In order to make

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

Bacterial amylases operate at higher temperatures than do fungal amylases. Fungal amylases react rapidly at lower temperatures; fungal amylases are used as an agent for alcohol fermentation for grain (Underkofler et al, 1958). Fungal amylases is said to be denatured – change shape (Alberte et al, 2012), at high temperatures above 60° C and bacterial amylases on the other hand are stable and show little denaturing at temperatures up to 85°C 3 The question answered by the experiment is if the temperature is not within the range of the enzymes (fungal and bacterial amylase) optimal temperature (higher temperature) then will the enzymes denature and if the enzymes are placed in lower temperature from optimal the activity then will it slow down enough to stop all reaction, meaning each enzyme will not be operating efficiently. Knowing about a bacterial amylases and fungal amylases optimal temperatures are important for knowing which food products and industrial products it can be used on to conserve the product because then the producer knows about which products it can be incorporated into depending on the temperature it is manufactured at.

The effects of temperature on fungal amylase Aspergillus oryzae, and bacterial amylase, Bacillus licheniformis ability to break down starch into maltose was studied. The study determined the optimal temperature the Aspergillus oryzae and Bacillus licheniformis was able to break down the fastest. The starch catalysis was monitored by an Iodine test, a substance that turns blue-black in the presence of starch. Amylase catabolizes starch polymers into smaller subunits. Most organisms use the saccharide as a food source and to store energy (Lab Manual, 51). The test tubes were labeled with a different temperature (0°C, 25°C, 55°C, 85°C). Each test tube was placed in its respective water baths for five minutes. After the equilibration process, starch was placed in the first row of the first row of the spot plate. Iodine was then added to the row revealing a blue black color. The starch was then added to the amylase. After every two minute section a pipette was used to transfer the starch-amylase solution to place three drops of the solution into the spot plate row under the corresponding temperature. Iodine drops was placed in the row. Color changes were noted and recorded. The results showed Aspergillus oryzae was found to have an optimal temperature between 25°C and 55°C and Bacillus licheniformis was found to have an

Hypothesis: If we decrease the level of pH in the enzyme Amylase, it will not be able to denature the carbohydrates in the potato starch solution after 10 drops because enzymes are very sensitive to pH levels and lowering it too much will compromise its ability to break them down.

The optimal temperature range of bacterial amylase, Aspergillus oryzae, were found and compared to the optimal temperature range of fungal amylase, Bacillus licheniformis, by testing the ability of the enzymes to hydrolyze starch after being exposed to different temperatures. The starch hydrolyzing ability of both the fungal amylase and the bacterial amylase was determined by mixing both amylases with starch and placing several tubes of the respective mixtures in four different temperatures each. Over the span of minutes, samples would be taken from each of the tubes and would be mixed with iodine. Due to the tendency of starch to color into a dark blackish-blue when combined with iodine, the degree of starch hydrolysis was then determined using a color coding scheme. The optimal temperature for both Aspergillus oryzae amylase and Bacillus licheniformis amylase was found to be within the 25 degree to 55 degree Celsius range. More specifically, the bacterial amylase seemed to hydrolyze starch more effectively as it spent more time exposed to the 55 degree Celsius temperature, whereas the fungal amylase seemed to work the best mostly when it had been exposed to the 55 degree Celsius temperature for 8 minutes. Within the resulting

In this lab our group observed the role of pancreatic amylase in the digestion of starch and the optimum temperature and pH that affects this enzyme. Enzymes are located inside of cells that increase the rate of a chemical reaction (Cooper, 2000). Most enzymes function in a narrow range of pH between 5 through 9 (Won-Park, Zipp, 2000). The temperature for which enzymes can function is limited as well ranging from 0 degrees Celsius (melting point) to 100 degrees Celsius (boiling point)(Won-Park, Zipp, 2000). When the temperature varies in range it can affect the enzyme either by affecting the constant of the reaction rate or by thermal denturization of the particular enzyme (Won-Park, Zipp, 2000). In this lab in particular the enzyme, which was of concern, was pancreatic amylase. This type of amylase comes from and is secreted from the pancreas to digest starch to break it down into a more simple form called maltose. Maltose is a disaccharide composed of two monosaccharides of glucose. The presence of glucose in our experiment can be identified by Benedicts solution, which shows that the reducing of sugars has taken place. If positive the solution will turn into a murky reddish color, where if it is negative it will stay clear in our reaction. We can also test if no reduction of sugars takes place by an iodine test. If starch is present the test will show a dark black color (Ophardt, 2003).

The Effects of Enzyme Concentration on the Activity of Amylase To investigate the effect of Amylase concentration on its activity. the relative activity of Amylase is found by noting the time taken for the starch substrate to be broken down, that is, when it is no longer gives a blue-black colour when tested with iodine solution. This time is referred to as the achromatic point. Equipment: v Amylase solution 0.1% v Starch Solution 1.0% v Distilled water v Iodine in potassium iodine solution v White tile and polythene pipette v Graduate pipettes or syringes v Test tubes in rack v Beaker (used as water bath) v Stopwatch, Thermometer v Eye Protection

In this lab we looked at the role of pancreatic amylase in the digestion of starch and the effect that temperature and pH has on this enzyme. Enzyme’s work as catalysts that increase the rate of chemical reactions within cells (Cooper, 2000). In order to do this, enzymes must show two essential properties: these two fundamental properties of enzymes include increasing the rate of chemical reactions without being eternally altered by the reaction and accelerating the reaction rate with keeping the reactants and products in chemical equilibrium (Cooper, 2000). Enzymatic catalysis is necessary for life. Most biochemical reactions would not occur under the mild temperatures and pressures