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Characterization of the Corynebacterium pseudotuberculosis enzyme CP40

Andersson Bennig, Isabella (2023) MOBY10 20222
Degree Projects in Molecular Biology
Popular Abstract
Amazing scissors or revolutionizing treatments?

Autoimmune diseases, cancer and allergies are something that is on the rise in the Western world and can cause major problems in everyday life, in the worst case even death. In addition to that, we have a growing problem with antibiotic-resistant bacteria and increased complications for both healthy people, and especially for those who already have a weakened immune system. Therefore, the research behind new drugs and therapies is very important. Current treatment involves the majority receiving tablets, intravenous medicine or injections for their ailments. Recently there has been a growth in use of antibody therapy. Antibodies are proteins that act as weapons, they can recognize... (More)
Amazing scissors or revolutionizing treatments?

Autoimmune diseases, cancer and allergies are something that is on the rise in the Western world and can cause major problems in everyday life, in the worst case even death. In addition to that, we have a growing problem with antibiotic-resistant bacteria and increased complications for both healthy people, and especially for those who already have a weakened immune system. Therefore, the research behind new drugs and therapies is very important. Current treatment involves the majority receiving tablets, intravenous medicine or injections for their ailments. Recently there has been a growth in use of antibody therapy. Antibodies are proteins that act as weapons, they can recognize specific targets and activate the immune system. Endoglycosidases is the collective name for the enzymes studied in this project. These enzymes can remove sugar molecules found on the antibodies and affect their structure, function and stability. The antibodies can then target specific structures or cells and activate the immune system.

The main focus of this project is the C. pseudotuberculosis protein CP40, a bacterial endoglycosidase. We wanted to produce and confirm the activity of this protein but also create an inactive version of the protein to determine the position of the active site within the protein's structure. In this project, we have therefore recombinantly produced the protein in E. coli cells and then purified it. Subsequently, the activity has been compared between the active and inactive CP40, but also between CP40 and other previously described endoglycosidases; EndoSD, EndoS and EndoS2.

In this project we have successfully produced CP40, inactive CP40 and EndoSD. Furthermore, we looked into their activity and can conclude that we successfully created an inactive CP40 variant. Meanwhile the regular CP40 needs some more work due to lower activity. The EndoSD protein was more active when produced at lower temperatures.

CP40 is a much smaller protein than EndoS or EndoS2, which makes it a good candidate for therapeutics. Hopefully, in the near future CP40 will be fully described and used in engineering antibodies for different therapeutic applications.

Master’s Degree Project in Molecular Biology 30 credits 2023
Department of Biology, Lund University
Advisor: Lotta Happonen & Mattias Collin
Advisors Division of Infection Medicine, BMC, Lund University (Less)
Please use this url to cite or link to this publication:
author
Andersson Bennig, Isabella
supervisor
organization
course
MOBY10 20222
year
type
H1 - Master's Degree (One Year)
subject
language
English
id
9122571
date added to LUP
2023-06-09 13:47:48
date last changed
2023-06-09 13:47:48
@misc{9122571,
  author       = {{Andersson Bennig, Isabella}},
  language     = {{eng}},
  note         = {{Student Paper}},
  title        = {{Characterization of the Corynebacterium pseudotuberculosis enzyme CP40}},
  year         = {{2023}},
}