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- Title
Methanol feeding strategy design enhances recombinant human growth hormone production by Pichia pastoris.
- Authors
Güneş, Hande; Boy, Erdem; Ata, Özge; Zerze, Gül H; Çalık, Pınar; Özdamar, Tunçer H
- Abstract
The effects of designedmethanol and co-substrate feeding strategies on fed-batch recombinant human growth hormone (rhGH) production by Pichia pastoris hGH-Mut+ were investigated. RESULTS: The highest cell concentration was obtained as 157 g L-1 with the exponential feeding strategy where methanol was fed at a pre-calculated feeding rate based on pre-determined μ0 =0.03 h-1, and mannitol concentration was kept constant at 50 g L-1 for t=0-6 h. The highest rhGH concentration was obtained as 1.3 g L-1 at t=54 h with the methanol-stat feeding strategy, where no co-substrate was used and the methanol concentration was kept at 5 g L-1 in the medium during the production phase. Thereafter, in anothermethanol-stat feeding strategywith pulse-fed sorbitol, the highestrhGHconcentration was obtained as 1.2 g L-1 at t=42 h. CONCLUSION: The methanol-stat fed-batch method keeping methanol concentration constant at 5 g L-1 in the production medium increased rhGH production at least 2-fold compared with that of the exponential feeding strategies. In the designed methanol-stat feeding strategies, use of sorbitol as co-substrate decreased methanol demand of the cells 2-fold, increased the overall product yield on the substrate 1.6-fold, shortened the cultivation time by 12 h and increased the productivity. Thus, methanol-stat feeding with co-substrate sorbitol was found to be a favorable fed-batch strategy for rhGH production by Pichia pastoris under alcohol oxidase I promoter.
- Subjects
HUMAN growth hormone; METHANOL; PICHIA pastoris; SORBITOL; ALCOHOL oxidase; RECOMBINANT proteins
- Publication
Journal of Chemical Technology & Biotechnology, 2016, Vol 91, Issue 3, p664
- ISSN
0268-2575
- Publication type
Article
- DOI
10.1002/jctb.4619