The Effect of different Temperatures on the reaction rate of Amylase Introduction: Amylases are enzymes that breaks down starch which accelerates the hydrolysis of glycosidic bonds in polysaccharides (S. Sivaramakrishnan et al., 2006). They bond with certain substrates to effectively increase the reaction rate of a chemical experiment. Most enzymes are very specific, as in they would only bond with a distinct substrate for it to cause a certain reaction. This can be described with the key and lock analogy. Only one type of key, certain amylase, will be able to open a particular lock, substrate. Without amylase, a lot of the organs necessary for people will no longer be able to react as fast, like the digestion of food or respiration. It speeds …show more content…
Then we add about one milliliter of 7 buffer to each of the tubes. The first tube is put into the 80 degrees Celsius water bath. The second is put into the 37 degrees Celsius. The third was left out at room temperature which is around 22 degrees Celsius. And the last one is put into a tub of crushed ice at 0 degrees Celsius. Then get a round of new test tubes and label them 1A through 4A. Add one milliliter of amylase to each of the tubes. After that is done, place these numbers with their match. So 1A will be put into the 80 degrees Celsius bath with 1. 2A will be put into the 37 degrees Celsius with 2 and so on. There should be one tube of amylase and one tube of starch at each temperature station. Allow the eight tubes to stay in their environment for about 10 minutes to give them time to adjust to their perspective environment …show more content…
T., Vogelsong, K. M., Lu, Y., Ellman, A. B., & Hudgens, G. A. (1996). Salivary α-amylase as a measure of endogenous adrenergic activity. Clin Physiol Clinical Physiology, 16(4), 433-448. Hiromi, K., Takasaki, Y., & Ono, S. (1963). Kinetics of Hydrolytic Reaction Catalyzed by Crystalline Bacterial α-Amylase. III. The Influence of Temperature. Bulletin of the Chemical Society of Japan Bull. Chem. Soc. Jpn., 36(5), 563-569. Malhotra, R., Noorwez, S., & Satyanarayana, T. (2000). Production and partial characterization of thermostable and calcium-independent alpha-amylase of an extreme thermophile Bacillus thermooleovorans NP54. Letters in Applied Microbiology Lett Appl Microbiol, 31(5), 378-384. Sivaramakrishnan, S. (march 11.2006). ͢ᾀ- Amylases from Microbial Sources- An Overview on Recent Developments. Food Technology Biotechnology, 44(2), 173-184. Sudhan, H., & Priya, S. (2012). Isolation and Production Optimization of Amylase Producing Bacteria from Soil by Submerged Culture Method. Research Gate: Pharmaceutical Sciences, 1(2012), 8-10. Retrieved February 17,
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
The Effect of α-Amylase Concentration on the Rate of Starch Hydrolysis in a Porcine Pancreas
Specifically, alpha-amylase is produced by the salivary glands of Homo sapiens (Humans) as well as many other mammalian species and is encoded by the gene AMY1A (Tracey 2017, p.22). The enzyme alpha-amylase is able to uptake polysaccharides including starch and glycogen as a substrate then hydrolyze the alpha-1,4-glycosidic linkages that connect the monosaccharides together (Tracey 2017, p.37). This is the reason as to why salivary amylase is also referred to as alpha-amylase (Tracey 2017, p.22).
1. Gathered all required materials to designated lab bench. 2. Considered all safety precautions including the prevention of spilling water to avoid falls, handling glassware carefully to prevent shattering, avoiding long periods of working with warm water to avoid burns and avoiding the digestion/inhalation of by-products produced after the reaction (e.g. ethanol and carbon dioxide gas). 3.
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
To calculate the amylase activity in the germinating barley (2g), we needed to know the amount of 0.5% starch used in the reaction tube (5mg) and the time taken to reach the achromic point, to get the average amount of starch hydrolysed / min to reach achromic point for 1mL of Diluted Amylase Extract(DAE), thus, we could get average amount of starch hydrolysed / min to reach achromic point for 25mL of DAE, which was same as the average amount of starch hydrolysed / min to reach achromic point for 5mL of amylase, therefore, we got the average amount of starch hydrolysed / min to reach achromic point for the total volume of the filtrated, non-diluted amylase extract, with knowing the weight of barley grains/germinant used to make the extract, we could calculate the amylase activity as mg starch hydrolysed/min/g barley tissue.
Using a 5 mL pipet, exactly 5 mL of distilled water was added to each of ten test tubes. The level was marked with a masking tape and the water was poured out. The test tubes were marked to their assigned temperatures (45°C, 40°C, 35°C, 30°C and 27°C).
In testing the differences that pH and temperature place on the enzyme, it will be possible to deduct the changes in enzymatic activity because of their surroundings. By using starch, a molecule that is hydrolyzed by amylase, it is possible to see the disappearance of substrate and creation of product with the addition of Lugol’s iodine (Fox 64). Adding the starch to the enzyme solution in the experiment allows for comparison between differing pH levels as well as the different temperatures that one might find amylase. It will be possible to determine what temperature and pH level are most effective for the enzyme to function at by running these comparisons.
The purpose of this experimentation is to find the differences of the concentration of the enzyme amylase to determine what outcome of the changes will have on the rate of the reaction between starch and amylase. We hypothesizes that as the reaction temperature of amylase solution and starch solution rises the pH reaction rate of both solutions will rise. Materials: 6 test tubes Marker
Effect of varying Temperatures on Enzymatic Activity of Bacterial and Fungal Amylase and hydrolysis of Starch
For example, the source of two types of proteases are Bacillus alcalophilus and Bacillus lentus. The most common sources of amylases are the bacteria Bacillus subtilis, Bacillus amyloliquefaciens, and Bacillus licheniformis. On the other hand, lipases are currently being sourced from yeast, bacteria, fungus, and from mammals. Biotex is the main source of cellulases, and have been in use since 1987.
Bacillus subtilis, is a Gram-positive, catalase-positive bacterium, found in soil and the gastrointestinal tract of ruminants and humans. Due to its excellent fermentation properties, with high product yields, it is used to produce various enzymes, such as amylase and proteases (Maarten van Dijl; Hecker ,
To determine the most favorable temperature for the reaction of alpha amylase's enzymatic conversion of starch to glucose water baths of various temperatures were used. First a stock solution was prepared prior to the experiment; it included alpha-amylase, I2KI indicator and .0033 g/ml of starch. Thirty-five mL of this solution was then combined with 35mL of distilled water into an Erlenmeyer flask in order to create the reaction flask. The flask was then added into one of the various water bath temperatures, 15, 35, 45, 55, 65 and 70 C. The reaction flask needed to be in the water baths long enough for it to reach the required temperature before the enzyme was added. 0.1mL of I2KI indicator was then added to each of the twelve cuvettes which were used in the timed readings. Then 5mL of the solution from the reaction flask was transferred into one of the cuvettes which contained I2KI indicator. Next, the starch-iodine complexes' absorbance were read on the spectrophotometer and recorded onto the appropriate table. Then, 1mL of alpha amylase was added into the reaction flask to begin the reaction, once
Alpha amylase is a protein enzyme present in grains that have starch. It is also present in humans, most commonly in saliva (Murray). The main purpose for alpha amylase is to break down, or hydrolase, polysaccharides, such as starch (Murray). When alpha amylase breaks down starch and glycogen, they cause a chemical reaction that creates the sugars maltose and glucose. Alpha amylose is also