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- 저자 : Trond E. Ellingsen (검색결과 2 건)
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Anne Tondervik,Simone Balzer,Tone Haugen,Håvard Sletta,Marit Rode,Karine Lindmo,Trond E. Ellingsen,Trygve Brautaset 한국생물공학회 2013 Biotechnology and Bioprocess Engineering Vol.18 No.4
The Norwegian salmonid alphavirus (NSAV)infects farmed Atlantic salmon and rainbow trout, causes pancreas disease and leads to economic losses and fish health issues for the aquaculture. Vaccines are available,but recurring infection outbreaks at Norwegian fish farms have led to endeavours in finding solutions for increased prevention. The NSAV E1 and E2 envelope proteins are potential targets for production of recombinant subunit vaccines and for generation of antibodies for diagnostics. Efficient expression of target proteins is necessary for these applications, and here we present a new strategy for expressing this kinds of viral proteins. We show that 5'-terminal fusion of signal sequences OmpA and CSP to the e1 and e2 genes and removal of the C-terminal hydrophobic interaction and transmembrane domains of E1 and E2 leads to significantly increased expression levels. Recombinant Escherichia coli strains for high-level production of E1 and E2 harbouring these modifications were established using the inducible XylS/Pm expression cassette. Furthermore,reduction of temperature to 16oC after induction leads to 4-fold increase in production for E1, and under high-celldensity cultivations we obtained production levels up to 2.3 g/L. We also show that these proteins can be purified from inclusion bodies by affinity chromatography. This demonstrates the present approach as promising for large scale production of such viral proteins.
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Marit H. Stafsnes,Kjell D Josefsen,Trond E. Ellingsen,Geir Kildahl-Andersen,Svein Valla,Per Bruheim 한국미생물학회 2010 The journal of microbiology Vol.48 No.1
Microbial culture collections are important resources for isolation of natural compounds with novel properties. In this study, a culture collection of around 1,500 pigmented heterotrophic bacteria was established. The bacteria were isolated from the sea surface microlayer at different sampling sites along the mid-part of the Norwegian coast. The bacterial isolates produced pigments of various coloration (e.g. golden,yellow, red, pink and orange). Methanol extracts of sixteen isolates were characterized with LC-Diodearray-TOF mass spectrometry analysis. The number of pigments per isolate varied considerably, and a tentative identification of the pigments was performed based on UV-absorbance profile and molecular formula assignation based on the accurate mass determination. The LC-MS analyses revealed that most of the pigments probably were carotenoids. Furthermore, we developed a high throughput LC-MS method for characterization and screening of a larger sub-fraction (300 isolates) of the culture collection. The aim was to screen and identify bacterial isolates producing carotenoids that absorb light in the UVA-Blue light. Six of the bacterial strains were selected for detailed investigation, including 16s rRNA sequencing, preparative HPLC for purification of major carotenoids and subsequent structural elucidation with NMR. Among the identified carotenoids were zeaxanthin, nostoxanthin and sarcinaxanthin, some with novel glycosylation patterns.
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