Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo
- - - - -

How to calculate Vmax and Km from substrate concentration (enzyme reaction)?

enzyme kinetics Km Vmax enzymekinetics

  • Please log in to reply
3 replies to this topic

#1 AdamG

AdamG

    member

  • Members
  • Pip
  • 2 posts
0
Neutral

Posted 11 April 2018 - 04:24 AM

I would like to calculate Vmax and Km. I have an enzyme reaction, and as the reaction goes by samples were taken at 0,4,8,13,19, and at 28 minute. The substrate concentration (mM) was 12,10,8,6,4,2 respectively. How can I calculate Vmax and Km?

 

t (min)  S (mM)

0           12

4           10

8           8

13         6

19         4

28         2

 

t (min) 0;4; 8; 13; 19;28

S (mM)12;10;8; 6; 4;2

 


Edited by AdamG, 11 April 2018 - 04:28 AM.


#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 6,546 posts
550
Excellent

Posted 11 April 2018 - 05:47 AM

Plot the values, use the equations for the respective unknowns.  Have a look a Lineweaver-Burke plots...



#3 mdfenko

mdfenko

    an elder emeritus

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 3,279 posts
201
Excellent

Posted 11 April 2018 - 12:35 PM

from what i see, your data is showing how much substrate is left at each time point. this is not enough to determine max and km. to do that, you have to make a substrate saturation curve by running a series of assays with a range of substrate concentrations.

 

with that data you can determine the kinetic constants for your enzyme.

 

keep in mind, if your enzyme is not pure then your constants will not be accurate.


talent does what it can
genius does what it must
i do what i get paid to do

#4 mdfenko

mdfenko

    an elder emeritus

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 3,279 posts
201
Excellent

Posted 11 April 2018 - 12:36 PM

see my response in the other forum where this question was asked.


talent does what it can
genius does what it must
i do what i get paid to do





Also tagged with one or more of these keywords: enzyme, kinetics, Km, Vmax, enzymekinetics

Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.