Secreted production of an elastin-like polypeptide by Pichia pastoris.
Elastin-like polypeptides (ELPs) are biocompatible designer polypeptides with inverse temperature transition behavior in solution. They have a wide variety of possible applications and a potential medical importance. Currently, production of ELPs is done at lab scale in Escherichia coli shake flask cultures. With a view to future large scale production, we demonstrate secreted production of ELPs in methanol-induced fed-batch cultures of Pichia pastoris and purification directly from the culture medium. The production of ELPs by P. pastoris proved to be pH dependent within the experimental pH range of pH 3 to 7, as an increasing yield was found in cultures grown at higher pH. Because ELP produced at pH 7 was partly degraded, a pH optimum for production of ELP was found at pH 6 with a yield of 255 mg of purified intact ELP per liter of cell-free medium.
Schipperus
Roelof
R
Agrotechnology and Food Sciences Group, Wageningen University and Research center, Bornsesteeg 59, 6708PD Wageningen, The Netherlands.
Teeuwen
Rosalie L M
RL
Werten
Marc W T
MW
Eggink
Gerrit
G
de Wolf
Frits A
FA
D000595:Amino Acid Sequence; D001483:Base Sequence; D004271:DNA, Fungal / Q000235:genetics; D004355:Drug Stability; D004549:Elastin / Q000096:biosynthesis* / Q000235:genetics; D005656:Fungal Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D000432:Methanol / Q000494:pharmacology; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D010843:Pichia / Q000187:drug effects / Q000378:metabolism*; D014176:Protein Biosynthesis
D004271:DNA, Fungal; D005656:Fungal Proteins; D010455:Peptides; D004549:Elastin; D000432:Methanol