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.
País-Chanfrau
José M
JM
Center for Genetic Engineering and Biotechnology, P.O. Box 6162, Havana 10600, Havana, Cuba. jose.pais@cigb.edu.cu
García
Yuneski
Y
Licor
Lisandra
L
Besada
Vladimir
V
Castellanos-Serra
Lila
L
Cabello
Cecilia I
CI
Hernández
Lester
L
Mansur
Manuel
M
Plana
Liuba
L
Hidalgo
Abdel
A
Támbara
Yanet
Y
del C Abrahantes-Pérez
María
M
del Toro
Yoandris
Y
Valdés
Jorge
J
Martínez
Eduardo
E
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