4. Answer the following questions as they relate to Michaelis-Menten kinetics: a

Question

4. Answer the following questions as they relate to Michaelis-Menten kinetics: a) (2 pts) Which of the following statements about Michaelis-Menten kinetics is correct? A) Km, the Michaelis constant, is defined as the concentration of substrate required for the reaction to reach maximum velocity. B) Km, the Michaelis constant, is defined as the dissociation constant of the enzyme-substrate complex C) Km, the Michaelis constant, is expressed in terms of the reaction velocity. Km, the Michaelis constant, is a measure of the affinity the enzyme has for its substrate. b) (2 pts) Which of the following statements about Vmax is correct? A) Vmax is the reaction velocity when [S] Km max occurs because the enzyme becomes a limiting factor Vmax is the reaction velocity when [S

Explanation / Answer

4.a. - Km the michaelis constant is a measure of affinity the enzyme for ts substrate.

b. Vmax occurs because the enzyme becomes a limiting factor.

C. In experiment make a enzyme concentration of a reaction is constant say 5mM. Then add 1mM, 2mM, 3mM , 4mM up to 10mM of substrate in 96well plate. Add enzyme to the substarte at the above concentration and kept for 5min. Then using spectrophotometer note the absorbance value that directly related to the recction speed or Vo.
Increasing the concentration of substrate speed up the process of reaction and increase the rate of a reaction Vo.

D. Parabolic graph

E.    Vo=Vmax S / Km + S ----- (1)
    S = 1/20Km or Km = 1/20S = 0.05 S --- (2)
     Sub 2 in 1

      Vo = Vmax X S / [0.05S + S]
        = Vmax X S / [1.05S]
       5 = Vmax / 1.05
       Vmax = 5.05 umol/min

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