Structural bioinformatics-based protein engineering of thermo-stable PETase from Ideonella sakaiensis
| Autor: | Hye-Young Sagong, Seongjoon Joo, Hwaseok Hong, Kyung-Jin Kim, Hogyun Seo, Seul Hoo Lee, Hyeoncheol Francis Son |
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| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
0301 basic medicine Bioengineering medicine.disease_cause Protein Engineering 01 natural sciences Applied Microbiology and Biotechnology Biochemistry 03 medical and health sciences Structural bioinformatics Bacterial Proteins 010608 biotechnology Enzyme Stability medicine Binding site Burkholderiales chemistry.chemical_classification Binding Sites biology Polyethylene Terephthalates Temperature Substrate (chemistry) Computational Biology Protein engineering biology.organism_classification 030104 developmental biology Enzyme Biodegradation Environmental chemistry Mutation Degradation (geology) Ideonella sakaiensis Bacteria Biotechnology |
| Zdroj: | Enzyme and microbial technology. 141 |
| ISSN: | 1879-0909 |
| Popis: | Poly(ethylene terephthalate) (PET), a widely used plastic around the world, causes various environmental and health problems. Several groups have been extensively conducting research to solve these problems through enzymatic degradation of PET at high temperatures around 70 °C. Recently, Ideonella sakaiensis, a bacterium that degrades PET at mild temperatures, has been newly identified, and further protein engineering studies on the PET degrading enzyme from the organism (IsPETase) have also been conducted to overcome the low thermal stability of the enzyme. In this study, we performed structural bioinformatics-based protein engineering of IsPETase to optimize the substrate binding site of the enzyme and developed two variants, IsPETaseS242T and IsPETaseN246D, with higher enzymatic activity at both 25 and 37 °C compared with IsPETaseWT. We also developed the IsPETaseS121E/D186H/S242T/N246D variant by integrating the S242 T and N246D mutations into the previously reported IsPETaseS121E/D186H/R208A variant. At the 37 °C incubation, the quadruple variant maintained the PET degradation activity for 20 days, unlike IsPETaseWT that lost its activity within a day. Consequently, this study exhibited 58-fold increase in the activity compared with IsPETaseWT. |
| Databáze: | OpenAIRE |
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