Sequence and Features
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21COMPATIBLE WITH RFC
- 23COMPATIBLE WITH RFC
- 25COMPATIBLE WITH RFC
- 1000INCOMPATIBLE WITH RFCIllegal BsaI site found at 940
The enzymatic hydrolysis of PET often occurs in a low rate. To promote this reaction, a great number of theories emerged, one of which suggesting that the corresponding enzyme performs much more efficient when fused with hydrophobin. This theory was supported by the fact that these small cysteine-rich proteins of exclusively fungal origin called hydrophobins can naturally adsorb to hydrophobic surfaces and to interfaces between hydrophobic (air, oil, and wax) and hydrophilic (water and cell wall) phases, and eventually assists in targeting and binding the substrate PET, promoting the enzymatic activity.
Previous studies have already shown encouraging results that confirms this theory. Other PET hydrolases have been proved to have positive results having fused certain hydrophobins. In our construct, we fused PETase with inJanus, a class I hydrophobin, aiming to promote its enzymatic activity.
PETase was found from a kind of microorganism（Ideonella sakaiensis 201-F6）living on PET as the main carbon source. It can degrade macromolecular polymers into monomers.PETase is the only enzyme found in bacteria which can degrade PET.
Janus could be produced by filamentous fungi, such as Ascomycetes and Basidiomycetes, and their scientific name is hydrophobin. Many different aspects of fungal development have been attributed to Janus. For example, they are thought to play a role in the formation of aerial hyphae and fruiting bodies. One of the most important features of Janus is that they are able to assemble spontaneously into amphipathic monolayers at hydrophobic–hydrophilic interfaces.
There are two classes of Janus, which are divided by the stability of their self-assembly. inJanus from Grifola frondosa belongs to Class I. They could generate very insoluble assemblies, which can only be dissolved in strong acids such as trifluoroacetic acid or formic acid.
 Espino-Rammer L, Ribitsch D, Przylucka A, Marold A, Greimel KJ, Herrero Acero E, Guebitz GM, Kubicek CP, Druzhinina IS. 2013. Two novel class II hydrophobins from Trichoderma spp. stimulate enzymatic hydrolysis of poly(ethylene terephthalate) when expressed as fusion proteins. Appl Environ Microbiol 79:4230–4238. doi:.10.1128/AEM.01132-13
 Ribitsch D, Herrero Acero E, Przylucka A, Zitzenbacher S, Marold A, Gamerith C, Tscheließnig R, Jungbauer A, Rennhofer H, Lichtenegger H, Amenitsch H, Bonazza K, Kubicek CP, Druzhinina IS, Guebitz GM. 2015. Enhanced cutinase-catalyzed hydrolysis of polyethylene terephthalate by covalent fusion to hydrophobins. Appl Environ Microbiol 81:3586-3592. doi: 10.1128/AEM.04111-14.