Team:UC Davis/Project/Catalyst

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Team:UC Davis - 2012.igem.org


Modules

We have created several modules and biobrick parts for the degradation and utilization of PET.

LC-Cutinase and Initial PET Degradation
When looking for a catalyst capable of breaking down PET, we came across a paper that conducted a metagenomic analysis of leaf-branch compost, identified a cutinase homolog, and demonstrated its PET-degrading activity [1]. It was found that this catalyst broke PET down into two by-products: ethylene glycol and terepthalic acid (TPA).
(pathway.jpg)
We had the LC-Cutinase gene synthesized with a pelB leader sequence and a 6-his tag and have placed it in the following construct.
(construct.jpg)
We decided to use pelB as a leading sequence on the cutinase gene since it has been shown to direct the enzyme to the periplasmic space [1] . Once the enzyme is led towards the membrane, there is a leakage that helps it be secreted into the extracellular matrix [1]. We hoped that this sequence would help the enzyme be secreted so the PET would more easily be degraded. When we ordered the cutinase sequence, we added pelB to the front of the sequence, in hopes of repeating the secretion shown in previous results.

Ethylene Glycol Modules


References
[1] S. Sulaiman, S. Yamato, E. Kanaya, J. Kim, Y. Koga, K. Takano, S. Kanaya. "Isolation of a Novel Cutinase Homolog with Polyethylene Terephthalate-Degrading Activity from Leaf-Branch Compost by Using a Metagenomic Approach." Applied and Environment Microbiology, vol. 78 no. 5, pp. 1556-1562, March 2012.

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