ENZYME KINETICS
Explain the importance of standard reference
Standard reference is important to find the equation from the standard curve of absorbance. The equation are used for finding the initial starch concentration value from the stock.
What is the function of iodine solution?
For inorganic oxidation reduction reactions, iodine are used as indicator to follow the changes of iodide ion and iodine element. And the Soluble starch solution is added. Only iodine element in the presence of iodide ion will give the characteristic blue black colour solution. Neither iodine element alone nor iodide ions alone will give the colour result.
Explain why starch turned blue when reacted with iodine?
Amylose in starch responsible for formation of a deep blue colour if iodine presence. Iodine is not very soluble in water, therefore the iodine reagent are dissolving iodine in water when the presence of potassium iodide. The starch are mixed with iodine in water, then an intensely starch/iodine complex colour is formed .This makes a linear tri iodide ion complex which is soluble that slips into the coil of the starch. There is transfer of charge between the starch and iodine. The iodine/starch complex has energy level spacing for absorbing visible light that make the complex intense blue colour. The complex indicating that redox titrations involved. when there is excess oxidizing agent, the complex is blue but when there is excess reducing agent, the I5- breaks up into iodine and iodide and the colour disappears.
Provide the relationship between substrate concentration and enzyme reaction.
The reactants of enzyme that catalyse reactions is substrates. Each enzyme is quite specific that acting on a particular substrates to produce a particular products. Which is long chain amino acids bound together by peptide bonds. When the enzyme are kept constant and the substrate concentration then gradually increased, the reaction velocity will increase until it reaches a maximum. After this point level, the increases of substrate concentration will not increase the velocity. As the substrate concentration increases, the amount products produced in every successive tube also increase. Then the enzyme attains the saturation level, since there are no more reaction sites remaining for binding. So the enzyme can work at full capacity and its reaction rate is independent of substrate concentration. This is was explained by Michealis, that an enzyme catalyse reaction at varying substrate concentrations is diphasic which is at low substrate concentration on the active sites of enzyme are not occupied by substrate and the enzyme rate varies with the substrate molecules concentration. The absorbance of resultant solutions were read at 540nm. The intensity of colour depends on the concentration of the reducing sugars produce.
What is Km?
The relationship between rate of reaction and concentration of substrate depends on the affinity of the enzyme for its substrate are term for Km (Michaelis constant) of the enzyme, an inverse measure of affinity. The Enzyme with a high Km has a low affinity for its substrate, requires a greater concentration of substrate to achieve Vmax. An enzyme with a low Km relative to the physiological concentration of substrate, is normally saturated with substrate, and act at more or less constant rate, regardless of variations in the concentration of substrate within the range.
What is Vmax?
The rate of reaction when the enzyme is saturated with substrate is the maximum rate of reaction, Vmax (maximum velocity).
Conclude the effect of temperature on the production using amylase.
The effects on the activity of amylase breaking down starch by changing of the temperature of amylase and starch. If the temperature are too low, there is no noticeable reaction rate since the enzyme is operating at a temperature too below its optimum. Increase in the temperature of a system results from the increase of the kinetic energy system. When a molecule collides, the kinetic energy of the molecules is converted to chemical potential energy of the molecules. Thus greater kinetic energy of the molecules in a system will increases the chemical potential energy. When the temperature increases it is possible that more molecules per unit time reach the activation energy. Thus rate of the reaction are increases. To convert substrate into product, enzymes must collide and bind to the substrate at its active site. Increasing temperature will increase the number of collisions of enzyme and substrate per unit time. The thermal deactivation occur if temperature above 100C, as enough thermal energy break some of the intra molecular interactions between polar groups as well as the hydrophobic forces between the nonpolar group.
Share your learning experience here.
From this experiment we learn on how to construct a graph to find the Vmax and Km value from the graph. The Enzymes are catalysts and increase the speed of a chemical reaction without themselves undergoing any permanent chemical change. This experiment are based on the factor that effect the activity of enzyme. It has shown in the experiment that if the amount of the enzyme is kept constant and the substrate concentration is then gradually increased, the reaction velocity will increase until it reaches a maximum point. After this point, the increases in substrate concentration will not increase the velocity. when this maximum velocity had been reached, all of the available enzyme has been converted to enzyme substrate complex. The reaction rate increases with temperature to maximum level, then abruptly declines with further increase the temperature. Most enzymes rapidly become denatured at temperatures above 40°C, the enzyme determinations below that temperature. Over a period of time, the enzymes will be deactivated at moderate temperatures. Storage of enzymes at 5°C or below is generally the most suitable. But, Some of enzymes lose their activity when frozen. The ice bath is used to stop the reaction of enzyme. However, I think there is some error in the results because we are conducting the experiment in a group, so the measurement reading is not too accurate.
Standard reference is important to find the equation from the standard curve of absorbance. The equation are used for finding the initial starch concentration value from the stock.
What is the function of iodine solution?
For inorganic oxidation reduction reactions, iodine are used as indicator to follow the changes of iodide ion and iodine element. And the Soluble starch solution is added. Only iodine element in the presence of iodide ion will give the characteristic blue black colour solution. Neither iodine element alone nor iodide ions alone will give the colour result.
Explain why starch turned blue when reacted with iodine?
Amylose in starch responsible for formation of a deep blue colour if iodine presence. Iodine is not very soluble in water, therefore the iodine reagent are dissolving iodine in water when the presence of potassium iodide. The starch are mixed with iodine in water, then an intensely starch/iodine complex colour is formed .This makes a linear tri iodide ion complex which is soluble that slips into the coil of the starch. There is transfer of charge between the starch and iodine. The iodine/starch complex has energy level spacing for absorbing visible light that make the complex intense blue colour. The complex indicating that redox titrations involved. when there is excess oxidizing agent, the complex is blue but when there is excess reducing agent, the I5- breaks up into iodine and iodide and the colour disappears.
Provide the relationship between substrate concentration and enzyme reaction.
The reactants of enzyme that catalyse reactions is substrates. Each enzyme is quite specific that acting on a particular substrates to produce a particular products. Which is long chain amino acids bound together by peptide bonds. When the enzyme are kept constant and the substrate concentration then gradually increased, the reaction velocity will increase until it reaches a maximum. After this point level, the increases of substrate concentration will not increase the velocity. As the substrate concentration increases, the amount products produced in every successive tube also increase. Then the enzyme attains the saturation level, since there are no more reaction sites remaining for binding. So the enzyme can work at full capacity and its reaction rate is independent of substrate concentration. This is was explained by Michealis, that an enzyme catalyse reaction at varying substrate concentrations is diphasic which is at low substrate concentration on the active sites of enzyme are not occupied by substrate and the enzyme rate varies with the substrate molecules concentration. The absorbance of resultant solutions were read at 540nm. The intensity of colour depends on the concentration of the reducing sugars produce.
What is Km?
The relationship between rate of reaction and concentration of substrate depends on the affinity of the enzyme for its substrate are term for Km (Michaelis constant) of the enzyme, an inverse measure of affinity. The Enzyme with a high Km has a low affinity for its substrate, requires a greater concentration of substrate to achieve Vmax. An enzyme with a low Km relative to the physiological concentration of substrate, is normally saturated with substrate, and act at more or less constant rate, regardless of variations in the concentration of substrate within the range.
What is Vmax?
The rate of reaction when the enzyme is saturated with substrate is the maximum rate of reaction, Vmax (maximum velocity).
Conclude the effect of temperature on the production using amylase.
The effects on the activity of amylase breaking down starch by changing of the temperature of amylase and starch. If the temperature are too low, there is no noticeable reaction rate since the enzyme is operating at a temperature too below its optimum. Increase in the temperature of a system results from the increase of the kinetic energy system. When a molecule collides, the kinetic energy of the molecules is converted to chemical potential energy of the molecules. Thus greater kinetic energy of the molecules in a system will increases the chemical potential energy. When the temperature increases it is possible that more molecules per unit time reach the activation energy. Thus rate of the reaction are increases. To convert substrate into product, enzymes must collide and bind to the substrate at its active site. Increasing temperature will increase the number of collisions of enzyme and substrate per unit time. The thermal deactivation occur if temperature above 100C, as enough thermal energy break some of the intra molecular interactions between polar groups as well as the hydrophobic forces between the nonpolar group.
Share your learning experience here.
From this experiment we learn on how to construct a graph to find the Vmax and Km value from the graph. The Enzymes are catalysts and increase the speed of a chemical reaction without themselves undergoing any permanent chemical change. This experiment are based on the factor that effect the activity of enzyme. It has shown in the experiment that if the amount of the enzyme is kept constant and the substrate concentration is then gradually increased, the reaction velocity will increase until it reaches a maximum point. After this point, the increases in substrate concentration will not increase the velocity. when this maximum velocity had been reached, all of the available enzyme has been converted to enzyme substrate complex. The reaction rate increases with temperature to maximum level, then abruptly declines with further increase the temperature. Most enzymes rapidly become denatured at temperatures above 40°C, the enzyme determinations below that temperature. Over a period of time, the enzymes will be deactivated at moderate temperatures. Storage of enzymes at 5°C or below is generally the most suitable. But, Some of enzymes lose their activity when frozen. The ice bath is used to stop the reaction of enzyme. However, I think there is some error in the results because we are conducting the experiment in a group, so the measurement reading is not too accurate.