Question

Michaelis-Menten plot and Lineweaver-Burk plot calculations- use provided data to generate both M-M and L-B plots. Use scatter plots with markers on Excel to determine Vinas, KM, kcat, kcat/KM. The total enzyme concentration is 5 μM. Graphs can be 2 page. Must be computer generated with all axes labeled. Substrate (mM Vo (mM/s) 2.73 5.45 8.17 10.9 40.4 0.124 0.181 0.212 0.228 0.303

Answer 1

Sol.

Michaelis-Menten(M-M) equation :

V₀ = V_max [S] / ( K_M + [S] )

So , in M-M plot , we plot V₀ vs [S]

Lineweaver - Burk(L-B) equation :

1/V₀ = ( K_M / V_max )*1/[S] + 1/V_max

So , in L-B plot , we plot 1/V₀ vs 1/[S]

In L-B plot , equation of line ,

y = 13.77x +3.025

So , 1/V_max = 3.025

and , V_max = 1/3.025 = 0.33057 mM / s

Also , from equation of line ,

K_M / V_max = 13.77

K_M = 13.77*0.33057 = 4.55194 mM

Also , K_cat    = V_max / [E]

As total enzyme concentration = [E] = 5 M = 0.005 mM

So , K_cat    = 0.33057 / 0.005 = 66.114 s^-1

K_cat / K_M = 66.114 / 4.55194 = 14.52435 mM^-1 s^-1