Question

how do you create a Michealis-Menten plot and a Lineweaver- Burk plot  using the equation that is determine from a calibration curve of absorbance vs. time? How do you convert the absorbance determine into concentration?

Answer 1

This question is based on Enzyme catalysis which is related to Absorbance and concentration.

Absorbance may be converted into concentration by the formula which is as follows:

A = ɛ.l.c,

where

A =  the amount of light which is absorbed by the sample in a given wavelength,

ɛ = the molar absorptivity,

l = that distance which the light travels through the solution,

and

c = concentration of the absorbing species which is measured in per unit volume.

I am giving here all the information about michalis menton plot and lineweaver burk plot in terms of concentration as well as absorbance. Please find it as an image form.

• Absorbance vs. time plot gives a linear plot which indicates the zero order reaction and shows negative slope, that is (k = –slope).
• ln(Absorbance) vs. time plot gives  a linear plot which indicates a first order reaction and shows negative slope (k = –slope).
• 1/(Absorbance) vs. time plot gives a linear plot which indicates a second order reaction and give the positive slope.(k = slope).
• 
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  no LUT I= (Page no-03 K₂ [F] o [s] [s] + Ky tkz KI K [Eo] [s] [s] tkm. M = m c y (3) 15] + km gеtuр a gum Shweta lounas km = Rytkal - Ri Mate constant for CEST Mate constant Of LES] for & km-Michaelis Menten Loustant formation * when all the enzymes are active the enzymes Je = K₂ [Elo = 4max. Case I At low concentration of substrate. 157 ccc km M = K₂ [E], [s] - Imax (5] & [s]' km km Ist order oorto substrate Case - II At leigh conc. of substrate 15/>>> km 4= K2 [6].(57 S] I= K2(Ej] = um olmax Lexo order wir.to substrate ase-14 il km = [5] 4 = Kelto] [s] · 2257
  
  (page no-04 K2 [Elo M = ma 2 Imax žuo order Ist order cette [s] Hole veate are some time denoted by'r'r Different kind of plot :- D Lineweaver Back Plot :- (vs its ) M = K2 [E] (s] - Amar (st km + [s] Kont [s] km test Mmax ($) I = km el ime [s] td Imax olmax km - Intercept 4 = = 0
  
  (Page no-05] km crmax Amox rs J Km
  
  (Page no-06) Normally, Absorbance vs time plot as follows :- is (Absorbance) 0 600 700 100 200 300 400 500 time (second) # Michealis - Menten plot in terms of Absorbance us time will be same as above plot and it will give negative Slope and indicates a zero order reaction.
  
  Page no-07) Crenerally, Lineweaver Buck Plot is drawn day I l rate vs a substrate concentrations # In terms of Absorbance vs time, Lineweaver Buck plot will be drawn between 1 Absorbance us estime which will give positive slope and will indicate second order reaction - # Lineweaver Burk plot is as follows: Absorbante -K= slope time(s) Intercept