The effects of temperature on Aspergillus oryzae and , Bacillus licheniformis
Tamira Carey
PID: 4861587
Lab partners: Group 4
Lab Section: U29
ABSTRACT
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
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
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
Hydrolysis of starch for fungal amylase Aspergillus Oryzae and bacterial amylase Bacillus Licheniformis at different temperatures.
Effect of varying Temperatures on Enzymatic Activity of Bacterial and Fungal Amylase and hydrolysis of Starch
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.
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.
For the experiment, the changes of temperature on anaerobic fermentation the process in which cells undergo respiration without oxygen in Saccharomyces cerevisiae was observed. The purpose of this experiment was to test the effect of four different temperatures on the rate of carbon dioxide production in yeast by measuring the fermentation rate. Saccharomyces cereviviae, also known as Baker 's yeast, is a unicellular, eukaryotic sac fungus and is good for this experiment because of its characteristic of alcohol fermentation. It was hypothesized that fermentation increases with increased temperature to a point of 37°C; above that point, enzyme denaturing will occur and fermentation will decrease. The group was able to document the carbon dioxide production and mark each of the temperature intervals which were tested at temperatures 4°C (refrigerator temperature), 23°C (Room temperature), 37°C (Human body temperature) and 65° Celsius (Equal to 150°F). The experiment was conducted by pouring yeast solution with 2% glucose in fermentation tubes, placing the tubes in the appropriate incubation temperature, marking the rise of the gas bubbles in the fermentation tubes which indicated carbon dioxide production. The results of this experiment were not supported by the hypothesis, creating different results from what was predicted. It is important to understand the fermentation rate of yeast so
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
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
The results from this experiment show the effects that concentration, pH and temperature have on enzyme activity. In part A, three different concentrations of enzyme solutions were tested. The undiluted enzyme had the fastest enzyme activity, the 1:9 dilution had the slowest enzyme activity and the 1:3 dilution fell in between the undiluted one and the 1:9 dilution. This result showed that the undiluted enzyme converts the amylose at a faster rate. Increasing enzyme concentration will increase the rate of reaction because more active sites are available to bind to substrate and more enzymes will be colliding with substrate molecules (Berkson 1937).
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).
Investigate the effect of temperature change on the rate of hydrolysis of starch using the enzyme amylase. Amylase investigation --------------------- AIM: To investigate the effect of temperature change on the rate of hydrolysis of starch using the enzyme amylase. These are variables that may affect the rate of the reaction: * concentration of amylase * concentration of iodine * temperature conditions of reaction * pH conditions of reaction I intend to investigate the effect of temperature change on the rate of the hydrolysis of starch, using the enzyme amylase.
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
Experimental setup A napkin/paper was placed under the spot plates and across the top was written Temperature (0⁰,25⁰,55⁰,85⁰) and on the side was written Time (0,2,4,6,8,10).After that, 4 test tubes were labeled each with a different temperature (0⁰,25⁰,55⁰,85⁰), enzyme source (B –Bacterial and F –Fungal) and the group number. Another 4 test tubes were labeled these with different temperatures, enzyme source (B or F), group number and the letter S (for starch solution). Then, 5ml of 1.5% starch solution were added into each of the test tubes labeled S. Effect of Temperature on Amylase activity.
The purpose of this lab was to determine the optimal temperature and pH for bacterial amylase activity. A spectrophotometer, a laboratory instrument that measures the amount of light absorbed by the sample (Green et al. 2015), was used to record the percent transmittance of the various temperatures and pH levels. The percent transmittance was then used to create a visual of the optimum conditions of bacterial amylase activity (See Figures 1 and 2). After performing this experiment, one can conclude the optimum temperature to be approximately 38.5°C and the optimum pH level to be 7.