An enzyme is a accelerator that speeds up the rate of reactions by take downing the activation energy. Each enzyme works better under optimum conditions, which favor the most active form for the enzyme molecule. Enzyme and substrate concentration, temperature, and pH are environmental factors of import by bring forthing the most reaction rate. Besides, the different of factors are used to analyze the effects of catalase and human amylase. Gas force per unit area Sensor, Vernier Gas Pressure interface, and Logger Pro are used to roll up the force per unit area of O. At 300C and pH 7 of catalase, the concentration of substrates and enzyme addition and the rates of reaction besides rise. However, substrate concentration goes up at 400C and pH 7 of human amylase.
Introduction:
Enzymes are catalytic proteins, which control a chemical reaction by increasing the rate of a reaction without being consumed in the procedure ( Enzyme 2007 ) .The catalytic velocities up the chemical reaction by take downing the activation energy, which needed to interrupt the chemical bonds between reactants to unite with other substances. In reactions, the substances at the beginning of the procedure are called substrate and enzyme can covert substrate molecules to merchandise molecules. At take downing the activation energy barrier, the enzyme has specific substrates to absorb adequate energy to make the passage province ( Campbell 2008 ) .
Enzymes are really specific in environmental factors, which affect the reaction rate. The enzyme merely works on the substrate that fits the active site and no other ( Campbell 2008 ) . The enzyme binds to the substrate called the active site, which is made up aminic acids ( Enzyme 2007 ) . The substrates enter active site, and the enzyme alterations shape such that its active site enfolds the substrate and catalyse the reaction more easy. Furthermore, the more substrate molecules are available, the more frequently they entree the active sites ( Campbell 2008 ) . Enzyme works better under optimum conditions including enzyme concentration, substrate concentration, temperature, and pH because they favor the most active form for the enzyme molecule. Nevertheless, the enzyme will denature and go less efficient the rate of reaction when the conditions get utmost change ( Campbell 2008 ) .
Each enzyme has a different specific temperature, which affects on the rate of reaction. Most human enzymes have optimum temperature of about 35-400C. Substrates collide with active site, the kinetic energy of the molecules coverts to increase chemical possible energy ( Campbell 2008 ) . Therefore, the temperature increases to make the activation energy and the rate of an enzymatic reaction addition. In add-on, if the chemical possible energy rises greater, some of the weak bonds of 3-D form of the active protein are broken ( Enzyme 2007 ) . It will denature of the protein and demobilize the protein. Therefore, the rate of reaction lessening when there are excessively much heat. Merely as each enzyme has an optimum temperature, pH degree besides has an optimal of the scope pH 6-8.The pH affects the construction of enzymes by changing basic amino acids or ionisation of acidic ( Campbell 2008 ) . Changes of pH affect to 3-D form of the protein, and enzymes become denaturized.
Substrate concentration and enzyme concentration affect the rate of reaction of enzymes. When substrate concentration additions, it means that more substrate is added ; the reaction rate additions because of utilizing more the active site of the enzyme ( enzyme 2007 ) . However, when the active site of the substrate is reached at farther point, enzymes are saturated to restrict reaction rates. Therefore, when the substrate concentration is changeless, the rate of reaction of is changeless. However, when the substrates remain changeless, the enzyme concentration additions and the rate of reaction besides increases until certain restricting concentration.
In the experiment, by different environment factors catalase is used to the existent experiment and human amylase is used to the simulation experiment. Catalase is enzyme nowadays in all life cells. It decomposes hydrogen peroxide into O and H2O and protects cells ( ) . Amylase a digestive enzyme made chiefly by the pancreas and salivary secretory organs. The primary map of the enzyme amylase is to interrupt down starches in nutrient so that they can be used by the organic structure to trip specific chemical reactions ( Amylase tests.2006 ) . In add-on, both the salivary and pancreatic amylases are I±-amylases in human physiology ; I±-amylase is an enzyme that hydrolyses alpha-bonds of big alpha-linked polyoses such as amylum and animal starch ( Amylase tests.2006 ) . I±-amylase is predicted to work best in the human organic structure temperature of 37 A°C and pH from 5.6 to 6.9. If organic structure heat exceeds 37A°C by excessively much cells become impaired or for good damaged, at lower temperature metamorphosis decreases without lasting harm until ice crystals form in the cells. Besides, if pH is highly high or low, the activity will diminish for most enzymes. Catalase is predicted in the temperature of 37.50C, the pH under of 8, and the enzyme and substrate concentration is high.
Methods:
In this experiment, we tested catalase activity by utilizing a yeast solution to find the effects of enzyme concentration, substrate concentration, temperature, and pH. Furthermore, we measured the rate of chemical reaction by bring forthing the force per unit area of O and interrupting down of H peroxide.
First, we connected the Gas force per unit area Sensor to the Vernier Gas Pressure interface.Then we connected the Vernier Gas Pressure interface to the laptop. From the Biology with Computers booklet, we opened the file “ 06 Enzyme ( Pressure ) ” from Logger Pro plan. Then we used a clean big trial tubing and placed the enzyme solution at the really underside of the trial tubing. In add-on, we used an Erlenmeyer flask to maintain the trial tubing from traveling during the experiment. Besides, we used the gum elastic stopper to infix and make a tight seal onto the trial tubing, and the stopper valve was in the closed place. We drew up the substrate solution ( 3 % H2O2+ H2O ) into the syringe and connected the syringe to the gum elastic stopper assembly. Subsequently, we opened the valve of the syringe and injected the peroxide solution into the trial tubing and instantly closed the valve and clicked the cod button on the Logger Pro. While we waited three proceedingss to roll up informations, we did n’t twirl or travel the trial tubing. When informations aggregation had finished, we removed the gum elastic stopper assembly and discarded the contents of the trial tubing. Then, we selected experiment and stored latest tally in the Logger Pro package. We clicked on the graph where the informations values began to increase, dragged the mouse point to the point where the graph began to look non-linear, and clicked the Linear Fit button to a additive regression.For all the experiments, we used the same procedures.
For enzyme concentration, we used 15 milliliter of H2O and 15 milliliter of 3 % H peroxide. We placed one bead of barm at the underside of the trial tubing, and we drew up 6 milliliters of substrate solution. Following the above processs, we performed the experiment and determined the rate as the incline of the curves we generated during the experiment. We repeated the experiment utilizing different enzyme concentrations of two, three, four, and five beads and calculated the inclines as mentioned before. Therefore, we recorded the informations in table one.
Furthermore, for substrate concentration we added 1 milliliter of H2O with 5 milliliters of 3 % H peroxide. We placed three beads of enzyme solution at the really underside of the trial tubing, and we drew up the 6 milliliter of substate solution into the syringe from the beaker. In the same methods above, we performed the experiment and determined the rate as a incline of the curves during the experiment. We repeated the experiment utilizing different substrate concentrations of 2, 3, 4, and 5 milliliter of H2O and 4, 3, 2, and 1 milliliter of 3 % H peroxide. Consequently, we recorded the informations in table two.
Similarly, we used 3 milliliter of H2O with 3 milliliters of 3 % H peroxide for proving the consequence of temperature on the enzyme. We added three beads of barm at the underside of the trial tubing and sealed it with the gum elastic stopper assembly. Then we placed the trial tubing in the flask half full with ice H2O and waited for three proceedingss. Besides, at the same we placed syringe in ice for three proceedingss. We recorded the temperature from the thermometer placed in the ice. After the three minute period, we removed the syringe from the ice and connected it to the gum elastic stopper assembly, and we followed the general processs to find the rate of reaction. We repeated the experiment utilizing different temperature of room temperature, 300C H2O bath, 400C H2O bath, 500C H2O bath, and 600C H2O bath and calculated the inclines as mentioned before. Therefore, we determined the informations in table three.
Finally, we added 3mL of the pH 3 solution and 3 milliliter of 3 % H peroxide. We placed three beads of yeast solution at the really underside of the trial tubing, and we drew up the solutions into the syringe. In the same methods above, we performed the experiment and determined the rate as a incline of the curves during the experiment. We repeated the experiment utilizing different pH solutions of pH 5, pH 7, pH 9, and pH 11. As the consequence, we recorded the informations in table four.
Consequences:
Figure 1. Relationship between the rate of reaction and temperature for the human amylase. The information is collected from a fake experiment by utilizing the plan Enzyme Investigation. In this experiment, the rate of reaction of human amylase are based on the invariable of substrate concentration of 0.01 mole/L, enzyme concentration of 1.0 A-10-6 mole/L, and pH of 7.Human amylase ‘s optimum temperature is 400C.
Figure 2.Relationship between the rate of reaction and the consequence of pH for the human amylase. The information is collected from a fake experiment by utilizing the computing machine package Enzyme Investigations. While temperature at 250C, substrate concentration of 0.01 mole/L, and enzyme concentration of 1.0 A-10-6 mole/L remain changeless, pH alterations different degree from 1 to 14.
Figure3. Relationship between the rate of reaction and the consequence of substrate concentration for human amylase. The information is collected from a fake experiment by utilizing the computing machine package Enzyme Investigations. In this experiment, temperature at 400C, enzyme concentration of 1.0 A-10-6 mole/L, and pH of 7 remain changeless.
Figure4.Relationship between the rate of reaction and the consequence of temperature for the enzyme catalase by utilizing the computing machine package Logger Pro and determined on a three proceedingss period each test. Catalase ‘s optimum temperature is 300C. In this experiment, the rate of reaction is based on the invariable of substrate concentration of 0.5 milliliters and barm of 3 beads.
Figure 5. Relationship between the rate of reaction and the consequence of pH for enzyme catalase.The informations is collected from the existent experiment by utilizing the computing machine package Logger Pro and determined the experiment on a three proceedingss period each test. In this experiment, 3 % H2O2 of 3mL and barm of 3 beads remain changeless, but degree of pH varies from 3 to 11.
Figure 6. Relationship between the rate of reaction and the consequence of enzyme concentration for enzyme catalase.The informations is collected from the existent experiment by utilizing the computing machine package Logger Pro and determined on a three proceedingss period per test. In this experiment, 15mL of H2O and 15ml of 3 % H2O2 remain changeless, but more beads for enzyme solutions ( barm ) are added.
Figure 7. Relationship between the rate of reaction and the consequence of substrate concentration for the enzyme catalase.The informations is collected from the existent experiment by utilizing computing machine package Logger Pro and determined on a three proceedingss period. The effects are based on invariable of enzyme solutions of 3 beads.
Discussion:
