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 degrees 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.
Center for Genetic Engineering and Biotechnology, P.O. Box 6162, Havana 10600, Havana, Cuba. jose.pais@cigb.edu.cu
José M
JM
País-Chanfrau
Yuneski
Y
García
Lisandra
L
Licor
Vladimir
V
Besada
Lila
L
Castellanos-Serra
Cecilia I
CI
Cabello
Lester
L
Hernández
Manuel
M
Mansur
Liuba
L
Plana
Abdel
A
Hidalgo
Yanet
Y
Támbara
María
M
del C Abrahantes-Pérez
Yoandris
Y
del Toro
Jorge
J
Valdés
Eduardo
E
Martínez
D000645:Ammonium Sulfate / Q000494:pharmacology; D019149:Bioreactors / Q000382:microbiology*; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation / Q000187:drug effects; D004492:Edetic Acid / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D009584:Nitrogen / Q000378:metabolism*; D010843:Pichia / Q000187:drug effects / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D011384:Proinsulin / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D013696:Temperature
D011994:Recombinant Proteins; C060793:miniproinsulin; D011384:Proinsulin; D004492:Edetic Acid; D009584:Nitrogen; D000645:Ammonium Sulfate