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Topic: Plotting Lineweaver-Burke for Enzyme Catalysis of Tyrosine  (Read 5273 times)

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Offline sule21

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Plotting Lineweaver-Burke for Enzyme Catalysis of Tyrosine
« on: November 13, 2006, 04:46:20 PM »
hi everyone, i'm new to this forum so please don't lambast me for asking a possible stupid question, but here it goes:

i've got a lab report for Chemical Kinetics, and i need help with plotting a Lineweaver-Burke graph to determine the Vmax and 1/V for 6 assays of tyrosine conversion to melanin. 

from my spectroscopy readings, for one of my assays, i've been given:

slope = Km/Vmax = 0.1128 (from absorption vs. time plot)
intercept of absorption vs. time plot = 0.236
t = 444s
= 7.39x10^-4 mol/L

how to i take this information and get a value for 1/V???

we're supposed to calculate the theoretical value of V from Km and Vmax given on our formulated Lineweaver-Burke plot, and the from our experiment...

am i just using the equation:

1/V = Km/Vmax x 1/ + 1/Vmax

i plugged in the slope (Km/Vmax) and the 1/, but how do i get my values for 1/V and 1/Vmax????



also....could i get my desired values if i just take my slopes and put them in a 1/V vs. 1/ graph, and extrapolate???? b/c i've tried that and my line isn't straight, even though its supposed to be...

HELP :o
« Last Edit: November 13, 2006, 05:37:53 PM by sule21 »

Offline Yggdrasil

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Re: Plotting Lineweaver-Burke for Enzyme Catalysis of Tyrosine
« Reply #1 on: November 14, 2006, 12:58:00 AM »
What raw data are you measuring to get these values?

Offline sule21

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Re: Plotting Lineweaver-Burke for Enzyme Catalysis of Tyrosine
« Reply #2 on: November 14, 2006, 02:07:04 AM »
slope = Km/Vmax = 0.1128 (from absorption vs. time plot)
intercept of absorption vs. time plot = 0.236
t = 444s
= 7.39x10^-4 mol/L (concentration of tyrosine off the bottle)

this is the information i've acquired from the lab for one assay

Offline Yggdrasil

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Re: Plotting Lineweaver-Burke for Enzyme Catalysis of Tyrosine
« Reply #3 on: November 14, 2006, 03:44:21 AM »
slope = Km/Vmax = 0.1128 (from absorption vs. time plot)
intercept of absorption vs. time plot = 0.236
t = 444s
= 7.39x10^-4 mol/L (concentration of tyrosine off the bottle)

this is the information i've acquired from the lab for one assay

The slope of your absorption v. time plot will not be Km/Vmax.  The slope of this plot is your V.  Km/Vmax is the slope of the Lineweaver-burke plot (which plots the 1/V vs 1/ for many reactions).

Offline sule21

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Re: Plotting Lineweaver-Burke for Enzyme Catalysis of Tyrosine
« Reply #4 on: November 14, 2006, 04:05:54 PM »
slope = Km/Vmax = 0.1128 (from absorption vs. time plot)
intercept of absorption vs. time plot = 0.236
t = 444s
= 7.39x10^-4 mol/L (concentration of tyrosine off the bottle)

this is the information i've acquired from the lab for one assay

The slope of your absorption v. time plot will not be Km/Vmax.  The slope of this plot is your V.  Km/Vmax is the slope of the Lineweaver-burke plot (which plots the 1/V vs 1/ for many reactions).

thanx....i figured it out....albeit, after pulling an all-nighter trying every which way possible

i guess sleep is more important than i thought  when trying to focus and study :(

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