Team:UC Davis/Project/Catalyst

From 2012.igem.org

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We are looking to overproduce the LC-Cutinase gene in <i>E. Coli</i> so that we can further characterize its ability to degrade PET.
We are looking to overproduce the LC-Cutinase gene in <i>E. Coli</i> so that we can further characterize its ability to degrade PET.
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<br>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.
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Revision as of 18:46, 8 August 2012

Team:UC Davis - 2012.igem.org


Cutinase

The iGEM team at UC Davis is working to produce a catalyst capable of degrading the common plastic, PET. Further description to come.

Background
When looking for a way to degrade 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].

What we're doing
We are looking to overproduce the LC-Cutinase gene in E. Coli so that we can further characterize its ability to degrade PET.

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.

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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