Vmax values, as determined from the Lineweaver-Burk plot, for the uninhibited, half uninhibited, and inhibited enzymes were, 0.3647, 0.1262, and 0.3087 μmol/min respectively. The non-linear regression V¬max¬ values for the same enzyme were 0.3343 (9.09% error as compared to Lineweaver-Burk plot), 0.1264 (0.16% error), and 0.2694 μmol/min (14.6% error) respectively. The differences in the values are due to the presence of error introduced by a Lineweaver-Burk plot, where data points at higher and lower
On a Michaelis-Menten plot, the presence of a competitive inhibitor results in a change of the Km value, but not the Vmax value. With an Uncompetitive inhibitor both the Km and the Vmax are decreased. A Noncompetitive inhibitor decreases the value of Vmax, but doesn’t affect Km. The Lineweaver-Burk plot is the reciprocal of the Michaelis-Menten plot. A competitive inhibitor affects a Lineweaver-Burk plot by causing a right shift in the x-axis. With an uncompetitive
com). In order to observed and analyze the effects of changing mushroom Tyrosinase and L-dopa concentrations on reaction rates, and to figure out what type inhibitor of Cinnamic acid, we was used the data to construct the Michaelis-Menten and Lineweaver- Burk plots. The Michaelis-Menten is the common known models of enzyme kinetic. The model demonstrates how the reaction rate relates to the concentration of substrate and enzyme. The rate equation of these enzymes is characterized by a sigmoidal curve
scatter was used to plot the data for this graph. The data of I=5, 15, 20 was copied ( ctrl+C) and pasted on the graph also. The Title of the charge, x-axis, y-axis, and legend were added properly. This step was repeated for inhibitor 2 and 3.
An enzymatic reaction also known as enzyme kinetics involves enzymes which are catalysts which speed up a reaction without being used up itself and do not appear as reaction products. Kinetics measures the rate of a chemical reaction to help determine the concentration or quantity of the enzyme of interest. Km and Vmax are used as constants in any enzyme reactions. (1) Vmax is known as the maximum velocity at which the reaction can be catalysed. It is used to measure the enzymes concentration and
lactose while series C contained the 150mM lactose. The amounts of buffer, ONPG, and water added to each tube in each series are listed in detailed in tables 4, 6, and 8. The Lineweaver- Burk plot of each series was created using the same steps used in the second part of the experiment. Then, the obtained line equation from each plot was used to calculate
product [P]1. In this model, the breakdown of the ES complex to yield P is the overall rate-limiting step. Three assumptions of a Michaelis-Menton plot are that a specific [ES] complex in rapid equilibrium between [E] and [S] is a necessary intermediate, the amount of substrate is more than the amount of enzyme so the [S] remains constant, and that this plot follows steady state assumptions. Steady state assumptions states that the intermediate stays the same concentration even if the starting materials
and km for LDH are reported in the result section of this report. The Vmax and km obtained from the Michaelias Menten plot and lineweaver plots are different as expected. The estimation of Vmax and Km from Michaelias Menten plot is not accurate as double reciprocal plot because the Vmax is an asymptote in Michaelias Menten plot. However, the Vmax found from double reciprocal plot based of y intercept while Km is calculated based of the respective
clearly below the control group curve, which means that the enzymatic activity slows down in presence of NaCl. But, we also want to know if this particular inhibitor is a non-competitive or competitive one. The third graph (figure 3) is a lineweaver-burk plot and will represent the inverse graph of the figure 2 curves. This particular graph will convert the original data that was curves into linear correlation so that we are able to made some assumption on the characteristics of the
The rate law or rate equation for a chemical reaction is an equation that links the reaction rate with concentrations or pressures of reactants and constant parameters (normally rate coefficients and partial reaction orders).[1] For many reactions the rate is given by a power law such as where [A] and [B] express the concentration of the species A and B, respectively (usually in moles per liter (molarity, M)). The exponents x and y are the partial reaction orders and must be determined experimentally…………
The purpose of this experiment was to analyze the activity and kinetic properties of an enzyme derived from wheat germ. For the first part of the experiment, we had to prepare a standard curve for the reaction product by measuring the velocity of the reaction catalyzed by acid phosphate that is extracted from wheat germ. Different enzyme concentrations were tested to see how fast they reacted. In order to quantify the amount of nitrophenol generated, six standards were prepared containing known
1. Introduction 1.1. Aim 1.2. Theory and principles 1.3. Application of principles 2. Experimental 2.1. Table of list of materials 2.2. List of apparatus 2.3. Procedure 3. Data and calculations 3.1. Record of all relevant raw data 3.2. Calculations with statistical manipulations 4. Results and discussion 4.1. Interpretation of data and comparison of results with known values 4.2. Discussion of the significance of the results 4.3. Was the original aim achieved 4.4. Conclusion 5
the formation of enzyme-substrate complex. It is rather difficult to determine Vmax directly from plot of r against [S]. It is however, possible to rearrange the Michaelis-Menten equation (4) so to permit some alternative plots for easy determination of Vmax. Two of the best known methods which make use of rearranged equations are as follows: 1.The Lineweaver-Burk method: r = v =
In this study, a three step purification of alkaline phosphatase from non-pasteurized milk was reported. It included cream extraction, n-butanol treatment and acetone precipitation. Different parameters like buffer concentration, temperature, pH, substrate concentration, acetone and n-butanol treatment were optimized to maximize the enzyme activity. The enzyme was fruitfully purified upto homogeneity from the milk, with percentage recovery and fold purification of 56.17 and 17.67 respectively. The