Question

In: Biology

(14 marks) The following kinetic data were obtained for an enzyme in the absence of inhibitor...

The following kinetic data were obtained for an enzyme in the absence of inhibitor (1), and in the presence of an inhibitor at 5 mM concentration (2). Assume[ET] is the same in each experiment. [S] (mM) (1) v(µmol/mL sec) (2) v(µmol/mL sec) 1 12 4.3 2 20 8 4 29 14 8 35 21 12 40 26

[S] (mM)	(1) v(µmol/mL sec)	(2) v(µmol/mL sec)
1	12	4.3
2	20	8
4	29	14
8	35	21
12	40	26

a. Using a graphing program (excel or sigmaplot) construct a Lineweaver burke plot representing the uninhibited reaction and the inhibited reaction on the same plot (to submit your graphs please paste into either a word or a pdf document along with your solutions)

b. What is the equation for each of the lines? (on the graph)

c. Determine Vmax and Km for the enzyme.

d. Determine the type of inhibition and the KI for the inhibitor.

Expert Solution

Accoring to the Lineweaver Berk equation-

The y- axis is 1/v (1/rate) and x axis is 1/s (1/substrate)

Best-fit values	1 (without inhibitor)	2 (with inhibitor)
Slope	0.06314 ± 0.001473	0.2117 ± 0.001806
Y-intercept when X=0.0	0.01955 ± 0.0007614	0.02008 ± 0.0009332
X-intercept when Y=0.0	-0.3096	-0.09487

b) Equation of curve without inhibitor (v1)-

y= 0.06314x + 0.01955

Equation of curve with inhibitor (v2)-

y= 0.2117x + 0.02008

c) For 1- Without inhibitor-

Y-intercept= 1/Vmax = 0.01955

Thus Vmax= 1/0.01955 = 51.15 µmol/mL sec

X-intercept= -1/Km = -0.3096

Km= 3.23mM

For 2- With inhibitor-

Y-intercept= 1/Vmax = 0.02008

Thus Vmax= 1/0.02008 = 49.8 µmol/mL sec

X-intercept= -1/Km = -0.09487

Km= 10.54mM

d) Since the Vmax with inhibitor is lower than without inhibitor and the Km with inhibitor is more than Km without inhibitor, thus the type of inhibition is- Mixed inhibition

gladiator answered 4 months ago

The following kinetic data were obtained for an enzyme in the absence of inhibitor (1)

Enzymes Kinetics:The following kinetic data were obtained for an enzyme in the absence of inhibitor (1), and in the presence of two different inhibitors (2) and (3) at 5 mM concentration. Assume [Etotal] is the same in each experiment. [S](mM) 1 ν(μmol/mL. sec) 2 ν(μmol/mL. sec) 3 ν(μmol/mL. sec) 1 12 4.3 5.5 2 20 8 9 4 29 14 13 5 35 21 16 12 40 26 18 a. From a double-reciprocal plot of the data, Determine Vmax...

3. The kinetics of an enzyme are studied in the absence and presence of an inhibitor...

3. The kinetics of an enzyme are studied in the absence and presence of an inhibitor (A). The intial rate is given as a function of substrat concentration in the table. V0 V0 [S] (mmol/L) no Inhibitor Inhibitor A 1.25 1.72 0.98 1.67 2.04 1.17 2.5 2.63 1.47 5.00 3.33 1.96 10.00 4.17 2.38 (a) What kind oof competitor is inhibition is involved (competitive, noncompetitive, uncompetitive)? [Please Provide Graph] (b) Determine Vmax and Km in the absence and presence of...

1. The following data were obtained from 3 separate enzyme kinetic experiments using 3 different substrates...

1. The following data were obtained from 3 separate enzyme kinetic experiments using 3 different substrates S1, S2 and S3 forming products P1, P2 and P3 respectively. The amount of enzyme in each reaction is 1 µM. Find out the rate and graph the data using a Michaelis-Menton and Lineweaver-Burk plots and determine the values for Km, Vmax, Kcat, and Kcat/Km. Which among the 3 substrate is best substrate for this enzyme and why? (4 points) [S] (mM) [P1] (mM)...

Students collected Beta-fructosidase enzyme activity data in the presence and absence of an unknown inhibitor. P-nitrophenol...

Students collected Beta-fructosidase enzyme activity data in the presence and absence of an unknown inhibitor. P-nitrophenol was used as a read out for enzyme activity. Quantity of product was determined from the absorbance of each sample after 5 minutes of reaction. Using the information, complete the table below and create a properly labeled Lineweaver burke plot with the appropriate axes and best fit lines for the data, in excel (graph is worth 6pts). Using this plot, calculate the approximate Vmax...

Consider the following data for an enzyme catalyzed hydrolysis reaction in the presence and absence of...

Consider the following data for an enzyme catalyzed hydrolysis reaction in the presence and absence of inhibitor I: [Substrate] M vo (µmol/min) voI (µmol/min) 6x10-6 20.8 4.2 1x10-5 29 5.8 2x10-5 45 9 6x10-5 67.6 13.6 1.8x10-4 87 16.2 Use the above data, do the following: a. Generate Lineweaver-Burk plots of the data. b. Explain the significance of the x-interecpt, y-intercept, and the slope. c. Identify the type of inhibition.

1.The following data were obtained in a study of an enzyme known to follow Michaelis-Menten kinetics:...

1.The following data were obtained in a study of an enzyme known to follow Michaelis-Menten kinetics: 2/27/ 20 V0 (mol/min) 217 325 433 488 647 Substrate added (mmol/L) 0.8 2 4 6 1,000 The Km for A) 1 mM., B) 1000mM, C) 2mM, D ) 4mM, E) 6mM this enzyme is approximately: 2. To A) the enzyme concentration. B) the initial velocity of the catalyzed reaction at [S] >> Km. C) the initial velocity of the catalyzed reaction at...

Below are shown three Lineweaver-Burk plots for enzyme reactions that have been carried out in the presence, or absence, of an inhibitor.

Below are shown three Lineweaver-Burk plots for enzyme reactions that have been carried out in the presence, or absence, of an inhibitor. Part A Indicate what type of inhibition is predicted based on Lineweaver-Burk plot. mixed inhibition competitive inhibition uncompetitive inhibition Part B Indicate which line corresponds to the reaction without inhibitor and which line corresponds to the reaction with inhibitor present. red - without inhibitor, blue - with inhibitor blue - without inhibitor, red - with inhibitor Part C...

(Chapter 14) The following data were obtained from a repeated-measures study comparing three treatment conditions. a....

(Chapter 14) The following data were obtained from a repeated-measures study comparing three treatment conditions. a. Use a repeated measures ANOVA with α=.05 to determine whether there are significant mean differences among the three treatments. (Hint: start by filling in the missing values on the table below). Be sure to show all formulas with symbols (and plug in numbers), steps, processes and calculations for all steps and all parts of all answers. b. Compute n2, the percentage of variance accounted...

For the reaction A + B + C → D The following data were obtained at...

For the reaction A + B + C → D The following data were obtained at constant temperature: Trial Initial [A] (mol/L) Initial [B] (mol/L) Initial [C] (mol/L) Initial Rate (mol/L . s) 1 0.2790 0.1670 0.0334 1.190 2 0.5580 0.1670 0.0334 2.381 3 0.5580 0.3340 0.0334 9.524 4 0.2790 0.1670 0.0668 1.190 1. What is the order with respect to each reactant? (Enter the order for A as Answer 1, that for B as Answer 2, and that for...

Two methods were used to measure fluorescence emission of a dye and following data were obtained;...

Two methods were used to measure fluorescence emission of a dye and following data were obtained; Method1 Method2 Mean 13.83 13.54 Standard deviation 0.288 0.375 # of measurements 6 6 A) Are the std. deviations are significantly different? B) Are the means are significantly different?