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Large-scale separation and production of engineered proteins, designed for facilitated recovery in detergent-based aqueous two-phase extraction systems

Selber, K; Tjerneld, Folke LU ; Collén, Anna LU ; Hyytia, T; Nakari-Setala, T; Bailey, M; Fagerstrom, R; Kan, J; van der Laan, J and Penttila, M, et al. (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)
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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
2007-07-06 15:03:51
date last changed
2017-07-30 03:48:09
@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},
  keyword      = {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},
  volume       = {39},
  year         = {2004},
}