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Title:Strategije statističnega načrtovanja eksperimentov pri kristalizaciji proteinov, primer lizocima
Authors:ID Jandrić, Jelenko (Author)
ID Tušar, Livija (Mentor) More about this mentor... New window
ID Turk, Dušan (Comentor)
Files:.pdf MAG_Jandric_Jelenko_2020.pdf (3,00 MB)
MD5: F2518DF2F0E27F6931E3CBE67CD25675
PID: 20.500.12556/dkum/57e69c4a-b8e8-44c0-bcf4-a59ca1cfaf7d
 
Language:Slovenian
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:FKBV - Faculty of Agriculture and Life Sciences
Abstract:Lizocim je encim, ki hidrolizira vezi med N-acetil muraminsko kislino in N-acetil-D-glukozaminskimi preostanki znotraj peptidoglikana v bakterijski celični steni, kar pomeni, da ima baktericidni učinek. Lizocim je naravno prisotna molekula v nekaterih živilih živalskega izvora in velikokrat deluje kot naravni konzervans. Namen raziskave je bil ugotoviti dejavnike, ki vplivajo na proces kristalizacije lizocima, oblikovati lestvico za oceno rezultatov kristalizacije ter na osnovi dobljenih načinov kristalizacije lizocima in izvedene raziskave oblikovati strategijo kristalizacije lizocima. Z uporabo delnega faktorskega načrta poskusov smo preučevali vplive treh soli in njihovih koncentracij, koncentracije glicerola, različnih vrednosti pH, temperature, načinov mešanja vzorcev in različnih koncentracij lizocima na njegovo kristalizacijo. Nastali kristali so bili slikani z robotskim sistemom za slikanje kristalov, rezultati uspešnosti kristalizacije pa so bili ovrednoteni po lastni lestvici. Uporabili smo linearno regresijo za ugotavljanje statistično značilnih povezav in metodo podpornih vektorjev za napovedovanje kristalizacije. Pokazali smo, da višje koncentracije NaCl in PEG 4000, nižji pH in nižje temperature, višje koncentracije proteina v raztopini vodijo do kristalizacije lizocima. Model podpornih vektorjev se je izkazal pri napovedovanju kristalizacije – površina pod ROC krivuljo je bila 0,84. Z uporabo eksperimentalnih načrtov, statistične analize opravljenih poskusov, linearne regresije in uporabo umetne inteligence lahko načrtujemo bolj optimalne korake kristalizacije proteinov.
Keywords:lizocim, kristalizacija, statistična analiza, načrtovanje eksperimentov
Place of publishing:Maribor
Year of publishing:2020
PID:20.500.12556/DKUM-78299 New window
NUK URN:URN:SI:UM:DK:M3K1ZNCP
Publication date in DKUM:04.01.2021
Views:1498
Downloads:122
Metadata:XML DC-XML DC-RDF
Categories:FKBV
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Licences

License:CC BY-NC-ND 4.0, Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
Link:http://creativecommons.org/licenses/by-nc-nd/4.0/
Description:The most restrictive Creative Commons license. This only allows people to download and share the work for no commercial gain and for no other purposes.
Licensing start date:02.12.2020

Secondary language

Language:English
Title:Strategies of statistic experimental design in protein crystallisation, example of lysozyme
Abstract:Lysozyme is an enzyme that hydrolyses bonds between N-acetyl muraminic acid and N-acetyl-D-glucosamine residues within the peptidoglycan in the bacterial cell wall, meaning it has a bactericidal effect. Lysozyme is a naturally occurring molecule in some foods of animal origin and often acts as a natural preservative. The purpose of the research was to determine the factors influencing the lysozyme crystallization process, to design a scale for evaluating the crystallization results and to form a lysozyme crystallization strategy based on the obtained lysozyme crystallization methods and the performed research. Using a partial factorial experimental design, the effects of three salts and their concentrations, glycerol concentrations, different pH values, the temperature, the way of sample mixing, and different lysozyme concentrations on its crystallization were studied. The resulting crystals were imaged with a robotic crystal imaging system, and the crystallization performance results.
Keywords:lysozyme, crystallization, statistical analysis, design of experiments


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