| 1. |
- Selber, K, et al.
(författare)
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Large-scale separation and production of engineered proteins, designed for facilitated recovery in detergent-based aqueous two-phase extraction systems
- 2004
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Ingår i: Process Biochemistry. - 1873-3298. ; 39:7, s. 889-896
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Tidskriftsartikel (refereegranskat)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
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| 2. |
- Baguma, Yona, et al.
(författare)
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Expression patterns of the gene encoding starch branching enzyme II in the storage roots of cassava (Manihot esculenta Crantz)
- 2003
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Ingår i: Plant Science. - 0168-9452 .- 1873-2259. ; 164:5, s. 833-839
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Tidskriftsartikel (refereegranskat)abstract
- Spatial and temporal expression patterns of the sbeII and sbeI genes, encoding starch branching enzyme II and I, respectively, in cassava (Manihot esculenta Crantz) were studied at different phenological stages of the crop. A partial cDNA for sbeII in cassava was cloned and used along with a cDNA-specific fragment of sbeI. As the cassava plant aged, the transcriptional activity of the sbeII and sbeI genes in the underground storage roots increased, whereas the activity in other organs remained the same or declined. At 180 days after planting (d.a.p.), levels of sbeII and sbeI transcripts in storage roots were very low, whereas at 360 d.a.p., the levels had increased dramatically. The 360 d.a.p. old storage roots also accumulated gbssII and gbssI transcripts, as well as a longer gbssI transcript, gbssI′. The difference between the gbssI and gbssI′ transcripts was shown to be due to differential splicing, whereby the gbssI′ transcript retained the first three introns. Unexpectedly, expression of sbeII and sbeI in the 360 d.a.p. storage roots exhibited fluctuations during the 24 h cycle, both under the normal light/dark regime and under continuous light or continuous dark conditions.
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