Effect of substrate concentration on enzyme activity

3 Pages
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Department
Chemical and Biochemical Engineering
Course Code
Chemical and Biochemical Engineering 2290A/B
Professor
Mita Ray

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Description
Week  4  :  Study  the  effect  of  substrate  concentration  on  enzyme  activity     Protocol:  You  are  studying  the  effect  of  substrate  concentration  on  rate  of  enzyme   activity.   Assay  Mixutre:   Kinetic  Assay:     3.9ml  Buffer  A   1ml  substrate   0.1ml  enzyme   Read  A410  nm  every  30  seconds  for  4  minutes   We’ve  provided  you  with  5  different  concentrations  of  substrate:     2.7mM,  1.35mM,  0.675mM  ,  0.3375mM,  0.16875mM   Time (min) [S]= [S] [S] [S] [S] 2.70mM =1.350mM =0.675mM =0.3375mM =0.16875m M 0 1 1.5 2 2.5 3 3.5 4     Calculation:   1. Calculate  the  km,  Vmax,  initial  velocity  –  graphically  using  Lineweaver  Burk   Plot,  Eddie  hofsteen  plot  &  Hanes  plot     2. Calculate  the  km,  Vmax  and  initial  velocity  –  using  equations.   3.     Discuss  :     1. What is the difference between a phosphatase and a phosphorylase?   2. What is a chromgenic substrate?   3. Write the equation of the reaction we are examining 4. Draw the structure of the substrate and reaction products
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