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Validation of a dispenser machine and enzyme kinetics for alcohol dehydrogenase

Andersson, Emelie LU (2018) KEMK03 20181
Department of Chemistry
Abstract
To optimize precision and accuracy of small dispensed volumes, a dispenser was built. A 96-well plate was loaded with four replicates of a linear pyranine dilution series to test the accuracy and precision of the dispenser machine. Three dilution series of pyranine (100-0 percent, 10-0 percent and 1-0 percent) were made. The results showed that the dispenser has very good accuracy and precision. The dispenser was then used in an enzyme kinetic experiment with alcohol dehydrogenase to reveal kinetic properties of this enzyme. A dilution series of the substrate ethanol was loaded onto a new 96-well plate to generate a Michaelis Menten curve. The kinetic parameters for the ethanol concentration 0,2 M and 0,1 M were as follows: KM 10,5 mM and... (More)
To optimize precision and accuracy of small dispensed volumes, a dispenser was built. A 96-well plate was loaded with four replicates of a linear pyranine dilution series to test the accuracy and precision of the dispenser machine. Three dilution series of pyranine (100-0 percent, 10-0 percent and 1-0 percent) were made. The results showed that the dispenser has very good accuracy and precision. The dispenser was then used in an enzyme kinetic experiment with alcohol dehydrogenase to reveal kinetic properties of this enzyme. A dilution series of the substrate ethanol was loaded onto a new 96-well plate to generate a Michaelis Menten curve. The kinetic parameters for the ethanol concentration 0,2 M and 0,1 M were as follows: KM 10,5 mM and KM 7,48 mM, kcat 1,06 s-1 and kcat 0,39 s-1. (Less)
Popular Abstract (Swedish)
Reproducerbarhet av experimentella resultat är centralt inom vetenskapliga metoder och ett standardsystem av enheter underlättar detta. Vetenskap använder ofta större och mindre måttenheter än de som används vardagligt. Det är svårt att behålla precision och noggrannhet när det kommer till att överföra väldigt små volymer, något man ofta gör i kemi. Mitt arbete har gått ut på att validera en dispensermaskins precision samt noggrannhet och sedan att med hjälp av denna undersöka kinetiska parametrar hos enzymet alkohol dehydrogenas. Resultaten visade att dispensermaskinen hade god reproducerbarhet med små spädningsserier. Genom att studera enzymets reaktionsförlopp, plottat som en så kallad Michaelis Menten kurva, kunde flera kinetiska... (More)
Reproducerbarhet av experimentella resultat är centralt inom vetenskapliga metoder och ett standardsystem av enheter underlättar detta. Vetenskap använder ofta större och mindre måttenheter än de som används vardagligt. Det är svårt att behålla precision och noggrannhet när det kommer till att överföra väldigt små volymer, något man ofta gör i kemi. Mitt arbete har gått ut på att validera en dispensermaskins precision samt noggrannhet och sedan att med hjälp av denna undersöka kinetiska parametrar hos enzymet alkohol dehydrogenas. Resultaten visade att dispensermaskinen hade god reproducerbarhet med små spädningsserier. Genom att studera enzymets reaktionsförlopp, plottat som en så kallad Michaelis Menten kurva, kunde flera kinetiska parametrar bestämmas. (Less)
Please use this url to cite or link to this publication:
author
Andersson, Emelie LU
supervisor
organization
course
KEMK03 20181
year
type
M2 - Bachelor Degree
subject
keywords
enzyme kinetics, dispenser machine, Gradis, alcohol dehydrogenase, biochemistry, biokemi
language
English
id
8951071
date added to LUP
2018-06-21 11:57:36
date last changed
2018-06-21 11:57:36
@misc{8951071,
  abstract     = {{To optimize precision and accuracy of small dispensed volumes, a dispenser was built. A 96-well plate was loaded with four replicates of a linear pyranine dilution series to test the accuracy and precision of the dispenser machine. Three dilution series of pyranine (100-0 percent, 10-0 percent and 1-0 percent) were made. The results showed that the dispenser has very good accuracy and precision. The dispenser was then used in an enzyme kinetic experiment with alcohol dehydrogenase to reveal kinetic properties of this enzyme. A dilution series of the substrate ethanol was loaded onto a new 96-well plate to generate a Michaelis Menten curve. The kinetic parameters for the ethanol concentration 0,2 M and 0,1 M were as follows: KM 10,5 mM and KM 7,48 mM, kcat 1,06 s-1 and kcat 0,39 s-1.}},
  author       = {{Andersson, Emelie}},
  language     = {{eng}},
  note         = {{Student Paper}},
  title        = {{Validation of a dispenser machine and enzyme kinetics for alcohol dehydrogenase}},
  year         = {{2018}},
}