Question

Name (Can be on graph paper or Excel. It is easier on Excel This is worth 10 points. 3 points for determining the correct data, 3 points for the graph, 2 points each for the Km and Vmax (be sure to remember units 1. Use this experimental data to plot a lineweaver burke graph 2. Use the graph determine them and Vmax of the reaction 3. Be sure to keep the same number of sig figs in your answer, and state the units of the answer 4. Put the answer in the yellow box Substrate UM VO nmol/min 0.41 0.52 0,66 4.16 6.66 KI remember units! max remember

Answer 1

Following is the - complete Answer -&- Explanation: for the given: Question in...typed & image format...

$\Rightarrow$Answer:

1.   K_m =  1.052 uM ( micro-mol/L )
2.   V_max =  10.42 nmol/min.  ( i.e. nano-mol/ minute )

$\Rightarrow$Explanation:

Following is the complete Explanation: for the above: Answer(s)...

• Given:

Following is the given Data, in the form of a table: prepared by using MS Excel, presented here..in...image format...

0.4 Substrate concentration, [S], uM Vo, (micro-mole/L) Umin. 1/[ S 1 2.86 2.5 0.52 3.34 1.923076923 0.66 4.16 1.515151515 0.5 /V 0.3496503 0.2994012 0.2403846 0.1501502 6.66

$\Rightarrow$Note: As we can see, in the above table: the given data: has been slightly modified: in order to prepare

the Lineweaver - Burk plot...

• Step - 1:

​​​​​​​Using the above data table: if we plot the data: i.e. 1/V_o vs. 1/ [S] , we will get the following Lineweaver - Burk plot...

prepared using MS Excel, presented here in...image format...

1/Vo vs. 1/[S] plot TO y = 0.101x + 0.096 28 1/Vo, min./nmol + 1/Vo vs. 1/[S] plot — Linear (1/Vo vs. 1/[S] plot) LILLO TOH 13 1/[S],L/ UM

$\Rightarrow$ Linear equation of the TrendLine: y = 0.101x + 0.096 ------------------------Equation - 1

$\Rightarrow$Where:

1. y = 1/V_o  
2.   x = 1/[S]
3. slope = K_m / V_max= 0.101
4. y - intercept = 1/ V_max= 0.096  
5. x - intercept = - 1 / K_m

• ​​​​​​​Step - 2:

We know, from:  Michaelis - Menten Equation: the following:

$\Rightarrow$ Michaelis Menten Equation:

$\Rightarrow$   1/V_o = K_m / V_max   x ( 1/ [S] ) +   1/ V_max----------------Equation - 2

Thus, if we plot: 1/V_o vs.  1/ [S] , from the plot: we will get the following :

i.e.

1. slope =  K_m / V_max
2. y - intercept =  1/ V_max
3. slope / y - intercept = K_m   ; i.e. Michaelis -Menten constant
4. 1/ y - intercept = V_max   ; i.e. Maximum reaction velocity

• Step - 3:

​​​​​​​Therefore: using the data: from Step - 1, we would get the following values:

1.   K_m =  slope / y - intercept = ( 0.101 ) / ( 0.096 ) =  1.052 uM ( micro-mol/L )
2.   V_max = 1/ y - intercept = 1 / ( 0.096 ) =  10.42 nmol/min.  ( i.e. nano-mol/ minute )

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