Large-scale separation and production of engineered proteins, designed for facilitated recovery in detergent-based aqueous two-phase extraction systems
| Autor: | Anna Collén, Merja Penttilä, Tiina Nakari-Setälä, Richard Fagerström, Joop van der Laan, M.-R. Kula, Folke Tjerneld, Michael Bailey, John Kan, Klaus Selber, Teppo Hyytiä |
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| Jazyk: | angličtina |
| Rok vydání: | 2004 |
| Předmět: |
Chromatography
Aqueous solution biology Hydrophobin Chemistry Extraction (chemistry) Large-scale extraction Bioengineering Cellulase biology.organism_classification Applied Microbiology and Biotechnology Biochemistry Fusion protein Partition coefficient Endoglucanase I Detergent-based aqueous two-phase systems Hydrophobin fusion protein Yield (chemistry) biology.protein Trichoderma reesei |
| Zdroj: | Selber, K, Tjerneld, F, Collén, A, Hyytiä, T, Nakari-Setälä, T, Bailey, M, Fagerström, R, Kan, J, Van Der Laan, J, Penttilä, M & Kula, M R 2004, ' Large-scale separation and production of engineered proteins, designed for facilitated recovery in detergent-based aqueous two-phase extraction systems ', Process Biochemistry, vol. 39, no. 7, pp. 889-896 . https://doi.org/10.1016/S0032-9592(03)00198-5 |
| DOI: | 10.1016/S0032-9592(03)00198-5 |
| Popis: | The feasibility and scalability of extraction in detergent-based aqueous two-phase systems for the separation of proteins from culture broth is demonstrated. At the same time the large-scale production of a fusion protein and the influence of cultivation scale on the efficiency of separation were investigated. An amphiphilic fusion protein EGIcore-HFBI was chosen, consisting of the catalytic core of the cellulase endoglucanase I and the small protein hydrophobin I, expressed homologously in Trichoderma reesei. Using the technical nonionic detergent Agrimul NRE 1205 the separation was successfully scaled up to 1200 l. No differences in yield or in partition coefficient were observed at 10 ml and 1200 l scale. Changes in the fermentation temperature and scale, however, can influence the properties of the protein and thus alter partition coefficient and yield. The decreased separation efficiency appears to be correlated with changes in glycosylation at lower cultivation temperatures. |
| Databáze: | OpenAIRE |
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