Recombinant expression, purification and biophysical characterisation of virulence factors from Gram-negative pathogens

Sampath Kumar Yalamanchili

Research output: ThesisDoctoral Thesis

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Abstract

Endotoxin are complex molecules that provide Gram-negative pathogens with antibiotic resistance. They contain a hydrophobic anchor called lipid A. Lipid A biosynthesis enzymes UDP-2,3 diacylglucosamine hydrolase (LpxH) and Lipid A disaccharide synthase (LpxB) have been identified as drug targets. Another enzyme target of importance is cholesterolalpha-glycosyltransferase (CGT) from H. pylori, which synthesises glucosyl-cholesterol protecting the bacterium from changing environmental conditions. Recombinant expression and biophysical studies on these enzymes indicates they are conformationally stable and monodisperse while bioinformatic studies predicted the membrane interaction region for substrate recognition. These studies lay the foundation for future work focussed on structure-based drug design.
Original languageEnglish
QualificationDoctor of Philosophy
Awarding Institution
  • The University of Western Australia
Award date5 Sep 2019
DOIs
Publication statusUnpublished - 2019

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Virulence Factors
Lipid A
Enzymes
Glycosyltransferases
Drug Design
Pylorus
Microbial Drug Resistance
Computational Biology
Endotoxins
Cholesterol
Bacteria
Membranes
Pharmaceutical Preparations

Cite this

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title = "Recombinant expression, purification and biophysical characterisation of virulence factors from Gram-negative pathogens",
abstract = "Endotoxin are complex molecules that provide Gram-negative pathogens with antibiotic resistance. They contain a hydrophobic anchor called lipid A. Lipid A biosynthesis enzymes UDP-2,3 diacylglucosamine hydrolase (LpxH) and Lipid A disaccharide synthase (LpxB) have been identified as drug targets. Another enzyme target of importance is cholesterolalpha-glycosyltransferase (CGT) from H. pylori, which synthesises glucosyl-cholesterol protecting the bacterium from changing environmental conditions. Recombinant expression and biophysical studies on these enzymes indicates they are conformationally stable and monodisperse while bioinformatic studies predicted the membrane interaction region for substrate recognition. These studies lay the foundation for future work focussed on structure-based drug design.",
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language = "English",
school = "The University of Western Australia",

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Recombinant expression, purification and biophysical characterisation of virulence factors from Gram-negative pathogens. / Yalamanchili, Sampath Kumar.

2019.

Research output: ThesisDoctoral Thesis

TY - THES

T1 - Recombinant expression, purification and biophysical characterisation of virulence factors from Gram-negative pathogens

AU - Yalamanchili, Sampath Kumar

PY - 2019

Y1 - 2019

N2 - Endotoxin are complex molecules that provide Gram-negative pathogens with antibiotic resistance. They contain a hydrophobic anchor called lipid A. Lipid A biosynthesis enzymes UDP-2,3 diacylglucosamine hydrolase (LpxH) and Lipid A disaccharide synthase (LpxB) have been identified as drug targets. Another enzyme target of importance is cholesterolalpha-glycosyltransferase (CGT) from H. pylori, which synthesises glucosyl-cholesterol protecting the bacterium from changing environmental conditions. Recombinant expression and biophysical studies on these enzymes indicates they are conformationally stable and monodisperse while bioinformatic studies predicted the membrane interaction region for substrate recognition. These studies lay the foundation for future work focussed on structure-based drug design.

AB - Endotoxin are complex molecules that provide Gram-negative pathogens with antibiotic resistance. They contain a hydrophobic anchor called lipid A. Lipid A biosynthesis enzymes UDP-2,3 diacylglucosamine hydrolase (LpxH) and Lipid A disaccharide synthase (LpxB) have been identified as drug targets. Another enzyme target of importance is cholesterolalpha-glycosyltransferase (CGT) from H. pylori, which synthesises glucosyl-cholesterol protecting the bacterium from changing environmental conditions. Recombinant expression and biophysical studies on these enzymes indicates they are conformationally stable and monodisperse while bioinformatic studies predicted the membrane interaction region for substrate recognition. These studies lay the foundation for future work focussed on structure-based drug design.

KW - lipid A biosynthesis

KW - bacterial virulence factors

KW - peripheral membrane protein

KW - cholesterol glycosylation

KW - recombinant protein compression and purification

KW - biophysical characterisation of proteins

KW - enzymes

U2 - 10.26182/5d8d633600ecc

DO - 10.26182/5d8d633600ecc

M3 - Doctoral Thesis

ER -