Optimization of thermophilictrans-isoprenyl diphosphate synthase expression inEscherichia coliby response surface methodology
| Autor: | Angelica A. Piccolomini, Matteo Borrotti, Davide De Lucrezia, Alex Fiabon |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Biomedical Engineering Bioengineering 02 engineering and technology Biology medicine.disease_cause Applied Microbiology and Biotechnology 03 medical and health sciences Drug Discovery medicine Response surface methodology Escherichia coli chemistry.chemical_classification ATP synthase Process Chemistry and Technology Thermophile General Medicine 021001 nanoscience & nanotechnology biology.organism_classification 030104 developmental biology Enzyme chemistry Biochemistry biology.protein Molecular Medicine Fermentation Heterologous expression Thermococcus 0210 nano-technology Biotechnology |
| Zdroj: | Biotechnology and Applied Biochemistry. 64:70-78 |
| ISSN: | 0885-4513 |
| Popis: | We optimized the heterologous expression of trans-isoprenyl diphosphate synthase (IDS), the key enzyme involved in the biosynthesis of trans-polyisoprene. trans-Polyisoprene is a particularly valuable compound due to its superior stiffness, excellent insulation, and low thermal expansion coefficient. Currently, trans-polyisoprene is mainly produced through chemical synthesis and no biotechnological processes have been established so far for its large-scale production. In this work, we employed D-optimal design and response surface methodology to optimize the expression of thermophilic enzymes IDS from Thermococcus kodakaraensis. The design of experiment took into account of six factors (preinduction cell density, inducer concentration, postinduction temperature, salt concentration, alternative carbon source, and protein inhibitor) and seven culture media (LB, NZCYM, TB, M9, Ec, Ac, and EDAVIS) at five different pH points. By screening only 109 experimental points, we were able to improve IDS production by 48% in close-batch fermentation. |
| Databáze: | OpenAIRE |
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