Large-scale separation and production of engineered proteins, designed for facilitated recovery in detergent-based aqueous two-phase extraction systems
(2004) In Process Biochemistry 39(7). p.889-896- Abstract
- 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... (More)
- 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 (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/122987
- author
- organization
- publishing date
- 2004
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Detergent-based aqueous two-phase systems, Large-scale extraction, Endoglucanase I, Hydrophobin fusion protein
- in
- Process Biochemistry
- volume
- 39
- issue
- 7
- pages
- 889 - 896
- publisher
- Elsevier
- external identifiers
-
- wos:000220422800013
- scopus:1542653049
- ISSN
- 1873-3298
- DOI
- 10.1016/S0032-9592(03)00198-5
- language
- English
- LU publication?
- yes
- id
- 78c39909-2e41-46bb-b1e2-e0151ae91e4e (old id 122987)
- date added to LUP
- 2016-04-01 12:20:00
- date last changed
- 2022-04-13 17:33:31
@article{78c39909-2e41-46bb-b1e2-e0151ae91e4e, abstract = {{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}}, author = {{Selber, K and Tjerneld, Folke and Collén, Anna and Hyytia, T and Nakari-Setala, T and Bailey, M and Fagerstrom, R and Kan, J and van der Laan, J and Penttila, M and Kula, M-R}}, issn = {{1873-3298}}, keywords = {{Detergent-based aqueous two-phase systems; Large-scale extraction; Endoglucanase I; Hydrophobin fusion protein}}, language = {{eng}}, number = {{7}}, pages = {{889--896}}, publisher = {{Elsevier}}, series = {{Process Biochemistry}}, title = {{Large-scale separation and production of engineered proteins, designed for facilitated recovery in detergent-based aqueous two-phase extraction systems}}, url = {{http://dx.doi.org/10.1016/S0032-9592(03)00198-5}}, doi = {{10.1016/S0032-9592(03)00198-5}}, volume = {{39}}, year = {{2004}}, }