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Improved yields of full-length functional human FGF1 can be achieved using the methylotrophic yeast Pichia pastoris

A Fantoni ; RM Bill ; Lena Gustafsson (Institutionen för kemi- och bioteknik, Molekylär bioteknik) ; Kristina Hedfalk (Institutionen för kemi- och bioteknik, Molekylär bioteknik)
Protein Expression and Purification (1046-5928). Vol. 52 (2007), 1, p. 31-39.
[Artikel, refereegranskad vetenskaplig]

We have produced human fibroblast growth factor 1 (hFGF1) in the methylotrophic yeast Pichia pastoris in order to obtain the large amounts of active protein required for subsequent functional and structural characterization. Four constructs were made to examine both intracellular and secreted expression, with variations in the location of the His6 tag at either end of the peptide. hFGF1 could be produced from all four constructs in shake flasks, but production was optimized by growing only the highest-yielding of these strains, which produced hFGF1 intracellularly, under tightly controlled conditions in a 3 L fermentor. One hundred and eight milligrams of pure protein was achieved per liter culture (corresponding to 0.68 mg of protein per gram of wet cells), the function of which was verified using NIH 3T3 cell cultures. This is a 30-fold improvement over previously reported yields of full-length hFGF1.

Nyckelord: Human fibroblast growth factor, Overproduction, Pichia pastoris, Purification



Denna post skapades 2007-02-07. Senast ändrad 2015-01-08.
CPL Pubid: 26365

 

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Institutioner (Chalmers)

Institutionen för kemi- och bioteknik, Molekylär bioteknik (2005-2007)

Ämnesområden

Cell- och molekylärbiologi

Chalmers infrastruktur