Thesis
Effect of oxoglutarate dehydrogenase complex insertion in cyanobacteria Synechocystis sp. PCC 6803
Washington State University
Master of Science (MS), Washington State University
2017
Handle:
https://hdl.handle.net/2376/104185
Abstract
Tricarboxylic acid (TCA) cycle is a sequence of catabolic reactions that play a part in NADH reductant production to supply cellular growth, homeostasis, repair system and cellular component biosynthesis. The TCA cycle of cyanobacteria is widely studied since it has been known as incomplete due to lack of 2-oxoglutarate dehydrogenase (OGDH) enzyme complex that converts 2-oxoglutarate to succinyl-CoA. The OGDH enzyme complex is a multi-enzyme complex that consists of three subunit enzyme components: 2-oxoglutarate dehydrogenase encoded by OGDH gene, dihydrolipoamide succinyltransferase encoded by SucB gene and lipoamide dehydrogenase encoded by LpdE gene. In this study, OGDH complex from Escherichia coli was introduced into the cyanobacteria Synechocystis sp. PCC 6803. Three heterologous genes that encoded OGDH enzyme complex were introduced into Synechocystis sp. PCC 6803 to convert 2- oxoglutarate to succinyl-CoA directly. Results demonstrate that OGDH gene was successfully inserted into Synechocystis sp. PCC 6803 while other two genes failed presumably due to lethal effects. The growth of CA1 strain, the mutant of Synechocystis sp. PCC 6803 containing OGDH gene, was compared to the wild type strain. Based on OD730 value, dry cell weight and chlorophyll a content under autotrophic and mixotrophic condition, there was no significant difference observed in all growth parameters between wild type strain and CA1 strain. It indicates that OGDH gene only, without other two genes, did not have measurable impact on the growth of Synechocystis sp. PCC 6803.
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Details
- Title
- Effect of oxoglutarate dehydrogenase complex insertion in cyanobacteria Synechocystis sp. PCC 6803
- Creators
- Cindy Angelia
- Contributors
- Shulin Chen (Degree Supervisor)
- Awarding Institution
- Washington State University
- Academic Unit
- Biological Systems Engineering, Department of
- Theses and Dissertations
- Master of Science (MS), Washington State University
- Publisher
- Washington State University; [Pullman, Washington] :
- Identifiers
- 99900525007901842
- Language
- English
- Resource Type
- Thesis