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Method development: Obtaining soluble Xantha-L

Andersson Bennig, Isabella (2021) MOBK01 20211
Degree Projects in Molecular Biology
Popular Abstract
Pure factory-made protein?

Chlorophyll does not only make plants green, but is also a vital part of the plant. It allows the usage of photosynthesis. The chlorophyll production has 20 steps, some more studied and some less. An important but yet not biochemically classified protein in the production of chlorophyll is Xantha-L. The protein forms one of the rings in the structure of chlorophyll and this happens in three steps. The protein and its’ steps are one of the most complex ones in the biosynthesis and it is hard to obtain soluble Xantha-L since it is a membrane bound protein.

The aim of this project is to develop a method in order to obtain as much Xantha-L as possible and measure its activity. To do so, I used my workers, a... (More)
Pure factory-made protein?

Chlorophyll does not only make plants green, but is also a vital part of the plant. It allows the usage of photosynthesis. The chlorophyll production has 20 steps, some more studied and some less. An important but yet not biochemically classified protein in the production of chlorophyll is Xantha-L. The protein forms one of the rings in the structure of chlorophyll and this happens in three steps. The protein and its’ steps are one of the most complex ones in the biosynthesis and it is hard to obtain soluble Xantha-L since it is a membrane bound protein.

The aim of this project is to develop a method in order to obtain as much Xantha-L as possible and measure its activity. To do so, I used my workers, a bacteria named Escherichia Coli. This bacteria was my factory workers and produced the protein of choice, in this case, Xantha-L and Ycf54 which is a helper protein that is needed for Xantha-L maturation. The protein sketch was given to the E.Coli as a piece of DNA. E.Coli produced the protein and after it was done, unfortunately, it did not get a paycheck for its hard work. Instead, I had to break the bacterias cell membrane in order to get out the content, in this case Xantha-L.

Since the liquid from the E.Coli contained a lot more than just Xantha-L, the protein and the debris had to be separated. This was done by putting the liquid in a tube that was able to sort Xantha-L away from the debris so all of the Xantha-L stayed in the tube while the debris just ran by. After that the contents of the tube were examined and separated by their molecular weight to confirm that it was really the correct protein that came out from the tube.

Soluble Xantha-L was obtained in small quantities during the different trials in this project. But there is definitely room for improvement and development of the purification methods in order to get bigger quantities of the protein.

Supervisor: Mats Hansson & David Stuart
Bachelor's thesis 15 hp - MOBK01
Department of Biology, Lund University (Less)
Please use this url to cite or link to this publication:
author
Andersson Bennig, Isabella
supervisor
organization
course
MOBK01 20211
year
type
M2 - Bachelor Degree
subject
language
English
id
9060084
date added to LUP
2021-06-30 15:50:32
date last changed
2021-06-30 15:50:32
@misc{9060084,
  author       = {{Andersson Bennig, Isabella}},
  language     = {{eng}},
  note         = {{Student Paper}},
  title        = {{Method development: Obtaining soluble Xantha-L}},
  year         = {{2021}},
}