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

Adele Fantoni, Roslyn M. Bill, Lena Gustafsson, Kristina Hedfalk*

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

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. © 2006 Elsevier Inc. All rights reserved.

Original languageEnglish
Pages (from-to)31-39
Number of pages9
JournalProtein Expression and Purification
Volume52
Issue number1
DOIs
Publication statusPublished - Mar 2007

Keywords

  • human fibroblast growth factor
  • overproduction
  • Pichia pastoris
  • purification

Fingerprint Dive into the research topics of 'Improved yields of full-length functional human FGF1 can be achieved using the methylotrophic yeast Pichia pastoris'. Together they form a unique fingerprint.

  • Cite this