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Introduction of effect of enzyme on amylase activity
Investigating the effects of different temperatures on amylase activity
Investigating the effects of different temperatures on amylase activity
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Recommended: Introduction of effect of enzyme on amylase activity
pH and temperature amylase activity from fungal and mammal
Introduction
The first enzyme that was produced industrially is amylase from a fungal source in 1894, it was used to treat digestive disorders. Amylase are groups of enzyme that breaks down starch into sugar and starts the process of chemical digestion. Its primary function is to digest enzymes and its optimum pH is 7. Amylase is measured by mixing a substrate with a buffer and measuring the change of the mixture. The reason why we measure amylase is to assist in diagnosis of different diseases such as abdominal pain (Reynolds, 2009). Starch breakdown of amylase has received a great deal of attention because of their technological significance and economic benefits, and is also
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Two spot plates were placed on a napkin that has Temperature 0˚, 25˚, 35˚, 50˚, 65˚, 90˚ Celsius. Three groups tested fungal amylase which is Alpha-amylase Aspergillus Oryzae and two groups tested mammal amylase which is Alpha-amylase from Porcine pancreas. Six test tubes were labeled with different temperature and enzyme source Mammal or Fungal Amylase. Add 2.5mL of 1% starch was added to each test tube. Afterwards, each test tube was placed into its respective temperatures. Another Six test tubes were obtained and were labeled with different temperatures and enzyme source Mammal or Fungal Amylase. Add 1ml of amylase Mammal or Fungal in each test tube and placed into its respective temperature. The test tubes were allowed to equilibrate for five minutes in their temperatures. After the calibration process, few drops of each solution were transferred from each test tube without removing the test tubes from their temperatures. These drops were added into the first row of wells on the spot plate, then add two drops of iodine to the wells on the spot plate and wait 1 minute. Use a color- coding scheme to convert the data to qualitative data into quantitative data this will serve as the control. Add 0.5 ml of amylase to the appropriate test tube containing starch and wait two minutes. Then add two drops of the starch-amylase mixture from each tube to the third row of …show more content…
Two spot plates were placed on a napkin that has pH 3, 4, 5, 6, 7, 8. Three groups tested fungal amylase which is Alpha-amylase Aspergillus Oryzae and two groups tested mammal amylase which is Alpha-amylase from Porcine pancreas. Six test tubes were labeled with different pH and enzyme source Mammal or Fungal Amylase. Add 1.5ml of starch and 1ml of the pH buffer solution to each test tube. Another six test tubes were labeled with different pH and enzyme source Mammal or Fungal Amylase. Add 0.5ml of amylase into the test tubes, and 0.5ml of pH buffer to each test tube. Allow the tubes to equilibrate for five minutes, then transfer a few drops of the starch solution from each pH treatment add them into the first row of wells on the spot plate. This serves as a control to make sure that the breakdown of starch is not affected by pH. Then add two drops of iodine reagent to each of these wells and wait one minute, then add 0.5ml of amylase to each of the appropriate tubes. After two minutes, add a few drops from each tube, and place in the corresponding wall. After collecting the data use a color coding scheme to convert the qualitative data to quantitative data. The numerical data from each group was used to calculate the mean starch concentration and the standard deviation. The mean and the standard deviation values were calculated after entering all the data into
Data from Table 1. confirms the theory that as the concentration of glucose increases so will the absorbance of the solution when examined with the glucose oxidase/horseradish peroxidase assay. Glucose within the context of this assay is determined by the amount of ferricyanide, determined by absornace, which is produced in a one to one ratio.1 Furthermore when examining the glucose standards, a linear calibration curve was able to be produced (shown as Figure 1). Noted the R2 value of the y = 1.808x - 0.0125 trend line is 0.9958, which is statistically considered linear. From this calibration curve the absorbance values of unknowns samples can be compared, and the correlated glucose concentration can then be approximated.
These labels indicated the lactose solution that was be placed into the mini-microfuge tubes. The varying lactose ph solutions were obtained. The four miniature pipets were then used, (one per solution,) to add 1mL of the solution to the corresponding mini-microfuge tubes. When this step is completed there were two mini-microfuge tubes that matched the paper towel. Then, once all of the solutions contained their respective lactose solutions, 0.5mL of the lactase enzyme suspension was added to the first mini-microfuge tube labeled LPH4 on the paper towel, and 4 on the microfuge tube. As soon as the lactase enzyme suspension was added to the mini-microfuge tube, the timer was started in stopwatch mode (increasing.) When the timer reached 7 minutes and 30 seconds, the glucose test strip was dipped into the created solution in the mini-microfuge tube for 2 seconds (keep timer going, as the timer is also needed for the glucose strip. Once the two seconds had elapsed, the test strip was immediately removed, and the excess solution was wiped gently on the side of the mini-microfuge tube. The timer was continued for 30 addition seconds. Once the timer reached 7:32 (the extra two seconds accounting for the glucose dip), the test strip was then compared the glucose test strip color chart that is found on the side of the glucose test strip
For example, incubating the samples at different temperatures would create more data points to establish an optimal temperature. From the results in the experiment in this study, it is known as temperature increases, enzymatic activity increase, and vise versa. However, what can not be observed is at what point does the increase in temperature begin to denature the enzyme, above 60°C. Furthermore, assays can be preformed to determine optimal pH, as well. From Dutta’s, and his partners, experiment it shows that there is a range where the Heliodiaptomus viduus’s lactase shows the most activity, which is between 5.0 and 6.0
For the first biuret test, approximately 10 drops of water were added to the test tube. After the water was inside of the test tube, 10 drops of 5M NaOH and 5 drops of the Biuret reagent solution was added. The test tube was then gently rolled by a member of the group to mix the solution and everyone recorded the color of the solution. The same procedure was followed to test for starch, protein, sugar, lipid, and the unknown, but instead of the initial 10 drops of water, a sample of the variable being tested was placed in the test tube.
For the heat inactivation, two sets of 11 tubes were set up. The indicated amounts of buffer, water, and ONPG listed in table 10 were added to each tube. In addition, the control enzyme (0.1ml) was added to each tube of the control set and the same amount of heated enzyme was added to each tube of the heated set. The absorbance readings were taken and recorded in table 10. Finally, two Lineweaver-Burk plots were created. The plot for the heated set is represented by graph 10 and graph 11 represents the control set. The Km and the Vmax for the heated set and the control set were determined.
Abstract: Enzymes are catalysts therefore we can state that they work to start a reaction or speed it up. The chemical transformed due to the enzyme (catalase) is known as the substrate. In this lab the chemical used was hydrogen peroxide because it can be broken down by catalase. The substrate in this lab would be hydrogen peroxide and the enzymes used will be catalase which is found in both potatoes and liver. This substrate will fill the active sites on the enzyme and the reaction will vary based on the concentration of both and the different factors in the experiment. Students placed either liver or potatoes in test tubes with the substrate and observed them at different temperatures as well as with different concentrations of the substrate. Upon reviewing observations, it can be concluded that liver contains the greater amount of catalase as its rates of reaction were greater than that of the potato.
The independent variable for this experiment is the enzyme concentration, and the range chosen is from 1% to 5% with the measurements of 1, 2, 4, and 5%. The dependant variable to be measured is the absorbance of the absorbance of the solution within a colorimeter, Equipments: Iodine solution: used to test for present of starch - Amylase solution - 1% starch solution - 1 pipette - 3 syringes - 8 test tubes – Stop clock - Water bath at 37oc - Distilled water- colorimeter Method: = == ==
However, the decrease varied depending on the temperature. The lowest temperature, 4 degrees Celsius, experienced a very low decrease of amylose percentage. Temperature at 22 degrees Celsius and 37 degrees Celsius, both had a drastic decrease in amylose percentage. While the highest temperature, 70 degrees Celsius, experienced an increase of amylose percentage. In conclusion, as the temperature increases the percentage of amylose decreases; however, if the temperature gets too high the percentage of amylose will begin to increase. The percentage of amylose increases at high temperatures because there is less enzyme activity at high temperatures. However, when the temperature is lower, more enzyme activity will be present, which results in the decrease of amylose percentage. This is why there is a decrease of amylose percentage in 4, 22, and 37 degrees Celsius. In this experiment the optimal temperature is 37 degrees Celsius, this is because this is the average human body temperature. Therefore, amylase works better at temperatures it is familiar
Purpose: This lab gives the idea about the enzyme. We will do two different experiments. Enzyme is a protein that made of strings of amino acids and it is helping to produce chemical reactions in the quickest way. In the first experiment, we are testing water, sucrose solution, salt solution, and hydrogen peroxide to see which can increase the bubbles. So we can understand that enzyme producing chemical reactions in the speed. In the second experiment, we are using temperature of room, boiling water, refrigerator, and freezer to see what will effect the enzyme.
at a volume of 4cm3. The preliminary work also proved to me that my basic method worked without any setbacks that may affect my results. Variables:.. The variables involved in the rate of reaction between amylase and starch are. The volume of amylase The volume of starch
Metabolism is extremely altered during the transition period. The prepartum is marked by an accentuated fetus growth (Bauman and Bruce Currie, 1980) and the onset of lactation; however, the postpartum is marked by an increase in milk production over time and deficit on the intake of nutrients. This situation causes a negative balance of nutrients, when the intake of nutrients does not meet the requirements of the animal; more specifically about energy, it is well-known that transition cows undergo negative energy balance (NEB), and it is easily characterized by body weight (BW) and body condition score (BCS) loss. Bertics et al., 1992, and Hayirli et al., 2002, observed that DMI is affected during the prepartum period, showing a decrease of about 30% on the last couple weeks before parturition. After parturition, DMI increases to support mainly the demands for lactation but it is still not enough to meet the animal demands (NRC, 2001). DMI is directly related to milk yield and strategies have been developed to maximize intake in order to increase milk production, this shows its importance by the fact of an increase of about 1 kg on milk production at peak of lactation reflects about 200 kg of extra milk on the current lactation. Providing quality feed and maximizing DMI during the transition period is of high importance in minimizing NEB and its detrimental effects consequently improving milk production. Metabolic demands are increased to support lactation, and homeorhetic adaptations occur to partition nutrients to the mammary gland. A good example it is the glucose metabolism, an important metabolite used as main fuel for cells and also transformed in lactose by the mammary gland, which is affected in order of trying to meet th...
The enzyme assay was repeated in water baths at four temperatures: an ice bath (approximately 4 degrees celsius), room temperature (approximately 23 degree celsius), 32 degree celsius, and 48 degree celsius. Test tube 9 was obtained and labeled 19. The appropriate solutions were added to each test tube. All tubes were preincubated at the appropriate temperature prior to the mixing of tubes. The tubes were then set aside to acclimate for 15 minutes. After the equilibrium was reached and the spectrophotometer was adjusted with the control (tube 1) the pairs 2 & 3, 4, & 5, 6 & 7, and 8, & 9 were mixed one at a time. The absorbance changes at 15 second intervals for 60 seconds for each temperature were
Investigating the Effect of Enzyme Concentration on the Hydrolysis of Starch with Amylase Aim: Investigate the effect of enzyme concentration on the rate of an enzyme-controlled reaction. Using amylase and starch as my example. Introduction: I am investigating the effect of the concentration of the enzyme, amylase on the time taken for the enzyme to fully breakdown the substrate, starch to a sugar solution. The varied variable will be the concentration and all other variables are going to be fixed. The different concentrations will be: 0.5% 0.75% 1.0% 1.5% 2% An enzyme is a class of protein, which acts as a biological catalyst to speed up the rate of reaction with its substrates.
The enzyme pancreas amylase causes the decomposition of starch. The starch during the chemical reaction broken down into disaccharides, lactase, sucrase, and maltase forms of pure sugar. Disaccharides are broken down to monosaccharides. Lactase changed into lactose, then into glucose and galactose sucrase changed sucrose into glucose and fructose these are all forms of sugars. These sugar may not all be utilized by the body. Maltase breaks down maltose 2 form molecules of glucose. Protein -stomach Pepcid and HCI break down proteins. These protein continue during the chemical reaction change to polypeptides. In the small intestines- Trypsin breaks down proteins and polypeptides to dipeptides. Then the dipeptides are changed into chymotrypsin decomposition of proteins and polypeptides to dipeptides. Carboxypeptidase breaks down polypeptides and dipeptides to amino acids. Aminopeptidase disintegrates of polypeptides & dipeptides to amino acids. Dipeptidase dissects of dipeptides to amino acids. Amino acids are more utilized by the digestive process; they are the building blocks of protein. Fats start the chemical digestive process in the mouth, this maybe because that many fats take longer to decompose. Lingual lipase has a minor role in beginning fat digestion. The stomach has an immense amount of chemical reaction going on at one time.
The exocrine function of the pancreas is that it produces enzymes that aids in the digestion of food. There are three important enzymes that are crucial in helping with digestion. The first digestive enzyme is amylase. Amylase function is to break down carbohydrates. The amylase enzyme is made in two places: the cells in the digestive tract that produces saliva and the main one specifically found in the pancreas that are called the pancreatic amylase (Marie, Joanne; Media Demand, “What Are the Functions of Amylase, Protease and Lipase Digestive Enzymes”). The amylase in the pancreas passes through the pancreatic duct to the small intestines. This amylase in the pancreas completes the process of digestion of carbohydrates. Consequently, this leads to the production of glucose that gets absorbed into the bloodstream and gets carried throughout the body. The next enzyme that aids in digestion of food is protease. While amylase breaks down carbohydrates, protease breaks down protein. Protease breaks down protein into the building block form of amino acids. The three main proteases that it produces are: pepsin, trypsin and chymotrypsin (Marie, Joanne; Media Demand, “What Are the Functions of Amylase, Protease and Lipase Digestive Enzymes”). Pepsin does not occur in the pancreas but it is the catalysis in starting the digestion of proteins. Trypsin and chymotrypsin are the two proteases that occur in