Use this URL to cite or link to this record in EThOS:
Title: Introduction of an alternative carbon fixation cycle into Synechocystis sp. PCC 6803
Author: Cotton, Charles
ISNI:       0000 0004 6496 1804
Awarding Body: Imperial College London
Current Institution: Imperial College London
Date of Award: 2017
Availability of Full Text:
Access from EThOS:
Access from Institution:
The aim of thesis was to replace the carbon-fixing Calvin cycle of Synechocystis sp. PCC 6803 with the 3-hydroxypropionate bi-cycle. A genome-scale metabolic model was made to test how integration of the 3-hydroxypropionate bi-cycle into the cyanobacterial cell might affect growth and central metabolism. Two separate approaches for knocking out the Calvin cycle were tested, by deleting either the phosphoribulokinase gene or replacing the RuBisCO operon with a metabolic bypass. The first DNA constructs for expression of all enzymes in the bi-cycle were made and introduced into Synechocystis sp. PCC 6803, although expression of proteins other than the malonyl-CoA reductase and propionyl-CoA synthase was not detected. Several enzymes of the bi-cycle were cloned and expressed in Escherichia coli for characterisation and protein crystallisation studies. The structure of the native state of the tri-functional malyl-CoA lyase involved in the 3-hydroxypropionate bi-cycle was solved. The efforts made in these studies should prove useful for the successful engineering of the remainder of the 3-hydroxypropionate bi-cycle into Synechocystis sp. PCC 6803.
Supervisor: Murray, James ; Rutherford, Bill Sponsor: Biotechnology and Biological Sciences Research Council
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral