Improved yields of full-length functional human FGF1 can be achieved using the methylotrophic yeast Pichia pastoris
Artikel i vetenskaplig tidskrift, 2007

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.

Overproduction

Purification

Pichia pastoris

Human fibroblast growth factor

Författare

A Fantoni

Universita degli Studi di Firenze

RM Bill

Aston University

Lena Gustafsson

Chalmers, Kemi- och bioteknik, Molekylär bioteknik

Kristina Hedfalk

Chalmers, Kemi- och bioteknik, Molekylär bioteknik

Protein Expression and Purification

1046-5928 (ISSN) 1096-0279 (eISSN)

Vol. 52 1 31-39

Ämneskategorier

Biokemi och molekylärbiologi

DOI

10.1016/j.pep.2006.10.014