View source for Part:BBa K2110800


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<partinfo>BBa_K2110800 short</partinfo>

This codon optimized sequence codes the wild type protein called PETase. PETase is a Poly(ethylene terephthalate) hydrolase, which was found in Ideonella sakaiensis (strain 201-F6) by Japanese scientists. It allows Ideonella sakaiensis to degrade PET. If properly transferred into engineered organisms as E.Coli or yeast, secreted PETase can catalyze the hydrolysis of PET to produce mono(2-hydroxyethyl) terephthalate (MHET) as the major product. Optimum temperature is 40 degrees Celsius for PET film hydrolysis and optimum pH is 9.


===Usage and Biology===
https://static.igem.org/mediawiki/parts/thumb/4/44/Prediction.png/569px-Prediction.png

https://static.igem.org/mediawiki/parts/5/5a/SecretionofPETase.jpeg

In order to build a PET degradation device, we had to increase the secretion of PET degradation enzymes. So we searched for a novel signal peptide candidate through literature search and determined the cleavage site using computational analysis.
<p>In order to release PETase (BBa_K2110800) from <em>E. coli</em>, the endogenous signal peptide was removed and a new signal peptide (NSP4) was attached. As a result, it can be seen that NSP4-PETase was secreted more into the media than PETase (BBa_K2110800).</p>

===References===
Han, S., Machhi, S., Berge, M., Xi, G., Linke, T., & Schoner, R. (2017). Novel signal peptides improve the secretion of recombinant Staphylococcus aureus Alpha toxin H35L in Escherichia coli. AMB Express, 7(1), 93.

===Sequence and Features===
<partinfo>BBa_K2110800 SequenceAndFeatures</partinfo>

Activity Assay<br/>
We detect the absorption peak of culture medium after culturing with PET film added for 2d. The absorption degree-wavelength curve is as follows.
https://static.igem.org/mediawiki/parts/thumb/6/60/T--Tianjin--partwt.png/800px-T--Tianjin--partwt.png

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===Functional Parameters===
<partinfo>BBa_K2110800 parameters</partinfo>
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