Elevated reaction temperatures are crucial for the efficient enzymatic
degradation of polyethylene terephthalate (PET). A disulfide bridge was
introduced to the polyester hydrolase TfCut2 to substitute its calcium binding site. The melting point of the resulting variant increased to 94.7°C (wild-type TfCut2: 69.8 °C) and its half-inactivation temperature to 84.6 °C (TfCut2: 67.3 °C). The variant D204C-E253C-D174R obtained by introducing further mutations at vicinal residues showed a temperature optimum between 75 and 80 °C compared to 65 and 70 °C of the wild-type enzyme. The variant caused a weight loss of PET films of 25.0 +/- 0.8% (TfCut2: 0.3 +/-0.1%) at 70 °C after a reaction time of 48 h. The results demonstrate that a highly efficient and calcium-independent thermostable polyester hydrolase can be obtained by replacing its calcium binding site with a disulfide bridge.
| Identifer | oai:union.ndltd.org:DRESDEN/oai:qucosa.de:bsz:15-qucosa-205582 |
| Date | 23 June 2016 |
| Creators | Then, Johannes, Wei, Ren, Oeser, Thorsten, Gerdts, André, Schmidt, Juliane, Barth, Markus, Zimmermann, Wolfgang |
| Contributors | Universität Leipzig, Fakultät für Biowissenschaften, Pharmazie und Psychologie, Elsevier, |
| Publisher | Universitätsbibliothek Leipzig |
| Source Sets | Hochschulschriftenserver (HSSS) der SLUB Dresden |
| Language | English |
| Detected Language | English |
| Type | doc-type:article |
| Format | application/pdf |
| Source | FEBS Open Bio 6 (2016) 425–432 doi: 10.1002/2211-5463.12053 |
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