Improving the expression of mini-proinsulin in Pichia pastoris

Increased expression of recombinant mini-proinsulin in Pichia pastoris in 2.5 l bioreactors was achieved by increasing the cultivation pH from 5.1 to 6.3, by decreasing the temperature from 28 to 22 °C, and by periodical addition of ammonium sulfate and EDTA to the culture broth. Using this procedur...

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Veröffentlicht in:	Biotechnology letters 2004-08, Vol.26 (16), p.1269-1272
Hauptverfasser:	Pais-Chanfrau, J.M, Garcia, Y, Licor, L, Besada, V, Castellanos-Serra, L, Cabello, C.I, Hernandez, L, Mansur, M, Plana, L, Hidalgo, A
Format:	Artikel
Sprache:	eng
Schlagworte:	Ammonium Ammonium Sulfate - pharmacology Biological and medical sciences Bioreactors Bioreactors - microbiology Biotechnology Cell Culture Techniques - methods Cell Proliferation - drug effects Edetic Acid - pharmacology Fundamental and applied biological sciences. Psychology gene expression humans Hydrogen-Ion Concentration insulin Microbiology Nitrogen - metabolism pH effects Pichia - drug effects Pichia - genetics Pichia - growth & development Pichia - metabolism Pichia pastoris proinsulin Proinsulin - biosynthesis Proinsulin - genetics protein expression systems Q1 Q2 recombinant proteins Recombinant Proteins - biosynthesis Temperature Temperature effects
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Zusammenfassung:	Increased expression of recombinant mini-proinsulin in Pichia pastoris in 2.5 l bioreactors was achieved by increasing the cultivation pH from 5.1 to 6.3, by decreasing the temperature from 28 to 22 °C, and by periodical addition of ammonium sulfate and EDTA to the culture broth. Using this procedure, mini-proinsulin reached nearly 0.3 g l-1 in the culture supernatant after 160 h of growth.
ISSN:	0141-5492 1573-6776
DOI:	10.1023/B:BILE.0000044871.80632.f9