(i) a) Construct an empty table with the following column headings: Substrate concentration [S] and initial velocity (Vi) where [S] has the unit µM, and Vi has the unit mM/s. (ii) The table provided is the enzyme kinetic data for your mutated enzyme, whereby Vi was expressed using the unit ∆A(405 nm)/s. Using the standard curve, express Vi with the unit mM/s rather than ∆A(405 nm)/s. Place your answer in the table above alongside the appropriate [S]. Hint: To answer this question you need to use this standard curve equation=0.0419x (The slope of the line is= 0.0419) (iii) The unmutated form of your protein has a Km of 25 µM and a Vmax of 43 mM/s. The enzyme kinetic data for your enzyme with the amino acid substitution should now be displayed in the table above. Based on these data, what is Vmax? Km? and explain the effect of amino acid substitution on the Km and Vmax for the mutated protein. (Up to 50 words) Hint: You are not expected to draw a Lineweaver-Burk plot, however a quick sketch of a Vi vs [S] plot may help you to answer the question. only correct answers pls!
|
SUBSTRATE CONCENTRATION [S] µM |
INITIAL VELOCITY V0 s-1 |
|
10 |
0.13 |
|
25 |
0.27 |
|
50 |
0.45 |
|
100 |
0.65 |
|
150 |
0.77 |
|
200 |
0.85 |
|
300 |
0.94 |
|
500 |
1.03 |
(i) a) Construct an empty table with the following column headings: Substrate concentration [S] and initial velocity (Vi) where [S] has the unit µM, and Vi has the unit mM/s.
(ii) The table provided is the enzyme kinetic data for your mutated enzyme, whereby Vi was expressed using the unit ∆A(405 nm)/s. Using the standard curve, express Vi with the unit mM/s rather than ∆A(405 nm)/s. Place your answer in the table above alongside the appropriate [S].
Hint: To answer this question you need to use this standard curve equation=0.0419x (The slope of the line is= 0.0419)
(iii) The unmutated form of your protein has a Km of 25 µM and a Vmax of 43 mM/s. The enzyme kinetic data for your enzyme with the amino acid substitution should now be displayed in the table above. Based on these data, what is Vmax? Km? and explain the effect of amino acid substitution on the Km and Vmax for the mutated protein. (Up to 50 words)
Hint: You are not expected to draw a Lineweaver-Burk plot, however a quick sketch of a Vi vs [S] plot may help you to answer the question.
only correct answers pls!
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