Lab Report On Enzyme Activity

About pH: If the pH level is less than 5, then the speed of the enzyme reaction will be slower.

This experiment involved the chosen enzyme, B-Galactosidase, to be tested with a substrate called o-nitrophenol-B-D-galactopyranoside (ONPG). The purpose was to determine over time the effects the enzyme had on the substrate concentration, as well as to examine the effect of lactose, a disaccharide on the formation of o-nitrophenol. The experiment utilized a spectrophotometer to determine at which the rate that the enzyme catalyzes, by timing the change in absorbance every 15 seconds, as well as observing any colour change. The amount of enzyme added to the B-Galactosidase is increased over time, and the ONPG is set to a constant value each trial. It was determined that through the trials of testing the absorbance of the enzyme, the faster

40 celcius. At lower temperatures, sucrase activity begins to be inactive but quickly becomes active. At higher temperatures, sucrase activity Laboratory Report/ Natalie Banc/ Enzyme Activity/ Elizabeth Kraske/ 09.22.2016/ Page [2] of [4] starts to slowly become less active. 4. Was sucrase activity higher at 25 °C or 55 °C? 55 degrees celcius Table 6: Effect of Sucrose Concentration on Sucrase Activity Optical Density 35 g/L 30 g/L 25 g/L 20 g/L 15 g/L 10 g/L 5 g/L 0 g/L 1 1.007 0.974 0.950 0.926 0.849 0.734 0.515 0.003 2 1.002 1.011 0.947 0.937 0.834 0.766 0.496 0.002 3 0.980 0.998 0.944 0.932 0.838 0.754 0.495 0.001 average 0.996 0.994 0.947 0.932 0.840 0.751 0.502 0.002 Effect of Sucrose Concentration on Sucrase Activity 5. State how sucrase activity changes with increasing sucrose concentration. First sucrase activity increases greatly. After 10 g/l sucrase activity continues to increase but at a slow rate until it reaches 30 g/l. At 30 g/l to 35 g/l sucrase activities mostly stayed the same

Lastly only germinating and whole barely seeds showed the presence of maltose, indicating only within these two there is amylase present which actively hydrolyzing starch into maltose, as per reaction 1 stated above. As dormant seeds have amylase concentrations that are far too low to be detected by this type of assay, as during dormancy energy demands for this state is considered to be zero. Hence the amylase concentration in the dormant seeds are far too low to be detected in the Benedict’s reagent test (Ernst,

The purpose of this experiment was to analyze the effects of the variables: temperature, pH, and enzyme concentration, on the enzymatic reaction rate of catalase and the level at which its products are released, measuring the rate of absorption using the indicator solution guaiacol and a spectrophotometer to develop a hypothesis of the ideal conditions for these reactions. My hypothesis is that the extremes in concentration, temperature and pH will negatively affect the Au rate. This experiment used 11 solutions contained in cuvettes. Each cuvette, once mixed, is placed in spectrophotometer and then a reading taken every 20 seconds. Cuvettes 1, 8, and 10 are used as blanks to zero out the spectrophotometer. They all lack the enzyme to help determine the absorption of just the enzyme.

In test tube B , dark-purple is given in test . It is different from the expected result. At 37℃, it should be a optimum temperature for the enzyme activity. There should not be the present of starch . Thus, there may be low amount of amylase in saliva which cannot break down the starch completely which give dark-purple colour . In test tube C , the iodine solution change from brown to dark blue which is different from the expected . It is because the amylase is denatured at 75℃ that the the activity of amylase is low or even stop. Therefore, the starch is not broken down into maltose by amylase. In the test D, a dark-brown solution is seen in the test tube after adding the iodine as the pH of the 1ml 0.5M HCl is not an optimum pH for the activity of amylase that the starch is broken down into maltose . Amylase may not break down the starch well. In test tube E, a colourless colour formed. It is because redox reaction occurred during the test. Idoine reduced into idoine ion ， which changre from brown to colourless. In test tube F, the iodine solution change from brown to purple . It is because the salt has a function of cofactor which will shorten the time for amylase to take to break down the

The purpose of quantitative analysis of protein using a spectrophotometer is to measure the concentration of proteins in a given sample. The experiment is conducted by laboratory method (Biuret Test) and using spectrophotometer to analyze the absorbance of reactants at 540 nm, hence determining the concentration of the proteins in a given sample.

Discuss in detail the general conditions necessary for affective enzyme action. Are the conditions the same for each enzyme? Why or why not? The best conditions are cold temperature, high concentration and a high pH.The conditions would be different for different enzymes because all proteins are different.

5 cm3 of starch solution were added into the 5 test tubes that were labeled test tubes.

In an organism 's body, chemical reactions are constantly taking place. These essential reactions can make or break the well-being of the body, yet the brain behind these changes is often times not recognized. This little brain or “macromolecule” is called an enzyme. An enzyme is a type protein that is able to speed up over 5,000 different reaction types an organism (2). Through catalyzation, the process of speeding up chemical reactions, enzymes attach to a substrate/molecule and break it down so that it can be used throughout the organism. Enzymes break down substrates in a very efficient way; through an assembly line (3). One enzyme starts off by attaching itself to a substrate at the active site, where the two undergo chemical reactions.

Both enzymes denatured at 85°C base on the black/blue liquid color in the spot plate, meaning that the temperature was above the temperature needed to function properly. At 0°C, the bacterial enzyme had the highest enzymatic activity, disproving the hypothesis. However, the fungal amylase had low enzymatic activity, it had the second darkest liquid color in the spot plate. At 25°C, both enzymes had the third darkest color, showing some starch breakage and enzyme activity. At 55°C, the bacterial amylase had the second highest enzyme activity, with the exception of the 0 minute mark, which just contains starch mix with iodine. The fungal amylase showed the highest enzyme activity was at this temperature, with the lightest color from all the temperatures. Thus the individual group data, showed bacterial amylase to have an optimal temperature at 0°C, and fungal amylase showed to have an optimal temperature at 55°C. Nevertheless, the class data does not support these findings by the individual group. Base on the class data, both bacterial amylase and fungal amylase have an optimal temperature of 55°C, proving the hypothesis. This difference between the class data and the individual data shows that errors must have occurred while preforming the bacterial enzyme

The effectiveness was based on the color scale. The color scale was based on amount of oxygen produced by the chemical reaction.The chemical guaiacol was used during the experiment to easily see the speed of the chemical reaction because guaiacol turns brown in the presence of oxygen. On average the pH 8 had a color level of 6.17 more at the 5 minute check than pH 3. This is because the acidic pH 3 denatured the enzyme and it was not able to bind with the substrate to produce oxygen therefore there was less of it. Another factor that can affect the chemical reaction is temperature. It can affect the enzymes by changing their speed and at a certain temperature denaturing them.When enzymes and substrates are moving quickly, they are more likely to collide and have a chemical reaction. The warmer the temperature the faster the chemical reaction is to a certain degree. The chemical reaction was the most effective at 40 because the enzymes and substrates were moving fast, but the temperature was not high enough to denature the enzyme. After 40 degrees, the temperature increase became harmful to the chemical reaction. The color scale at 40 was 9.58 and at 50 the scale was 8.12. This shows at what temperature the enzyme begins to denature. Cold temperatures slow chemical reactions. At 10 degrees, the reaction occurred slower and this can be shown by the data. 40 was

Enzymes are biological catalysts, which are essential for carrying metabolic reactions in the human body including the breakdown of food for digestion, absorption and energy production. All biological reactions within human cells depend on enzymes (Wolfenden 1). It is essential for humans to have well-functioning enzymes to break down large molecules into smaller units. As a matter of fact, in the absence of normal functioning enzymes, the human body would cease to exist because chemical reactions that are required to maintain the body function would not occur fast enough. I have a lot of interest in health and human nutrition. Therefore I wanted to examine the breakdown properties of a digestive enzyme while under the influence of a strong inhibitor. For my experiment I chose amylase as an enzyme and starch as a substrate (which is broken down into glucose by Amylase). I selected Copper Sulphate as enzyme inhibitor against the concentration of 2% of Amylase solution. Light absorbance was the method used to

ABSTRACT: The purpose of the experiments for week 5 and week 6 support each other in the further understanding of enzyme reactions. During week 5, the effects of a substrate and enzyme concentration on enzyme reaction rate was observed. Week 6, the effects of temperature and inhibitor on a reaction rate were monitored. For testing the effects of concentrations, we needed to use the table that was used in week 3, Cells. The 3 concentrations of enzymes were 0.5 ml, 1.0 ml, and 2.0 ml of turnip extract, while the substrate consisted of 0.1ml, 0.2 ml, and 0.4 ml of hydrogen peroxide. In a separate tube, the control was made up of turnip extract and guaiacol, known as the color reagent. This was recorded the absorbance every 20 seconds for 3 minutes.

The trials of timed intervals showed that the independent variable, time, is directly proportional to the dependent variable, amount of H2O2 used. Compared to the uncatalyzed reaction of H2O2, the enzyme-catalyzed reaction had a much faster rate of decomposition.