Oxygen-limited fed-batch process: an alternative control for Pichia pastoris recombinant protein processes.
An oxygen-limited fed-batch technique (OLFB) was compared to traditional methanol-limited fed-batch technique (MLFB) for the production of recombinant Thai Rosewood beta-glucosidase with Pichia pastoris. The degree of energy limitation, expressed as the relative rate of respiration (q(O)/q(O,max)), was kept similar in both the types of processes. Due to the higher driving force for oxygen transfer in the OLFB, the oxygen and methanol consumption rates were about 40% higher in the OLFB. The obligate aerobe P. pastoris responded to the severe oxygen limitation mainly by increased maintenance demand, measured as increased carbon dioxide production per methanol, but still somewhat higher cell density (5%) and higher product concentrations (16%) were obtained. The viability was similar, about 90-95%, in both process types, but the amount of total proteins released in the medium was much less in the OLFB processes resulting in substantially higher (64%) specific enzyme purity for input to the downstream processing.
Charoenrat
Theppanya
T
School of Biotechnology, Royal Institute of Technology, AlbaNova University Centre, Roslagstullsbacken 21, 106 91, Stockholm, Sweden.|School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima, 30000, Thailand.
Ketudat-Cairns
Mariena
M
School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima, 30000, Thailand.
Stendahl-Andersen
Helle
H
School of Biotechnology, Royal Institute of Technology, AlbaNova University Centre, Roslagstullsbacken 21, 106 91, Stockholm, Sweden.
Jahic
Mehmedalija
M
School of Biotechnology, Royal Institute of Technology, AlbaNova University Centre, Roslagstullsbacken 21, 106 91, Stockholm, Sweden.
Enfors
Sven-Olof
SO
School of Biotechnology, Royal Institute of Technology, AlbaNova University Centre, Roslagstullsbacken 21, 106 91, Stockholm, Sweden. enfors@biotech.kth.se.
D000465:Algorithms; D019149:Bioreactors / Q000382:microbiology*; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D003198:Computer Simulation; D035782:Dalbergia / Q000201:enzymology*; D005246:Feedback / Q000502:physiology; D000432:Methanol / Q000378:metabolism; D008954:Models, Biological; D010100:Oxygen / Q000378:metabolism*; D010843:Pichia / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis; D001617:beta-Glucosidase / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics
D011994:Recombinant Proteins; D001617:beta-Glucosidase; D010100:Oxygen; D000432:Methanol