Insight
Green Ocean is working with cyanobacteria commonly known as blue- green algae. Cyanobacteria can be found in many different environments, including freshwater and marine ecosystem. We decided to work with cyanobacteria because research has shown that cyanobacteria has the ability to export proteins out of their membrane. This is necessary for our project, as we aim in the SuperPETase enzyme to be exported out of the membrane into PET plastic, which in turn be degraded by this enzyme. TorA pathway is the pathway that will be used to export our enzyme.
We are currently working in genetically modify marine cyanobacteria which will enable it to degrade polyethylene terephthalate (PET), most common form of plastic in the oceans. The engineered cyanobacteria harbor PETase, an enzyme that breaks down PET.We have modified the prototypical Ideonella sakaiensis PETase gene to be compatible with expression and secretion and detection in cyanobacteria. This engineering was accomplished in a dual-host plasmid shuttle vector in E coli, which would then be transferred to a model cyanobacterium Synechococcus elongatus by conjugation. We have added Part BBa_K3103000 to the registry, this part contains the TorA signal sequence followed by SuperPETase sequence with the mutations necessary to optimize the PETase enzyme. We have successfully been able to grow and culture cyanobacteria in the lab and are currently working in the conjugation process of cyanobacteria to our desire gene.
Citations
- Borunda, Alejandra. “This Young Whale Died with 88 Pounds of Plastic in Its Stomach.” National Geographic, 22 Mar. 2019, https://www.nationalgeographic.com/environment/2019/03/whale-dies-88-pounds-plastic-philippines/.
- Hornigold, Thomas, et al. “How Cyanobacteria Could Help Save the Planet.” Singularity Hub, 31 Jan. 2019, https://singularityhub.com/2018/06/04/how-cyanobacteria-could-help-save-the-planet/.
- Long, Kat. “New Species of Bacteria Eats Plastic.” The Wall Street Journal, Dow Jones & Company, 10 Mar. 2016, https://www.wsj.com/articles/new-species-of-bacteria-eats-plastic-1457636401.
- Spence, Edward, et al. “Membrane-Specific Targeting of Green Fluorescent Protein by the Tat Pathway in the Cyanobacterium Synechocystis PCC6803.” Molecular Microbiology, vol. 48, no. 6, Dec. 2003, pp. 1481–1489., doi:10.1046/j.1365-2958.2003.03519.x.
- Whitaker, Hannah. “How the Plastic Bottle Went from Miracle Container to Hated Garbage.” National Geographic, 24 Aug. 2019, https://www.nationalgeographic.com/environment/2019/08/plastic-bottles/.
- Wüstneck, Bernd. “In a First, Microplastics Found in Human Poop.” National Geographic, 23 Oct. 2018, https://www.nationalgeographic.com/environment/2018/10/news-plastics-microplastics-human-feces/.
- Yoshida, Shosuke, et al. “A Bacterium That Degrades and Assimilates Poly(Ethylene Terephthalate).” Science, vol. 351, no. 6278, Oct. 2016, pp. 1196–1199., doi:10.1126/science.aad6359.