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)),...

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Bibliographic Details
Published in:Bioprocess and biosystems engineering 2005-10, Vol.27 (6), p.399-406
Main Authors: CHAROENRAT, Theppanya, KETUDAT-CAIRNS, Mariena, STENDAHL-ANDERSEN, Helle, JAHIC, Mehmedalija, ENFORS, Sven-Olof
Format: Article
Language:English
Subjects:
Algorithms
b-glucosidase
beta-Glucosidase - analysis
beta-Glucosidase - biosynthesis
beta-Glucosidase - genetics
Bioengineering
Biological and medical sciences
Bioreactors - microbiology
Biotechnology
Bioteknik
Carbon dioxide
Cell Culture Techniques - methods
Cell Proliferation
Computer Simulation
Dalbergia - enzymology
expression
Feedback - physiology
Fundamental and applied biological sciences. Psychology
growth
Methanol
Methanol - metabolism
methanol-limited fed batch (MLFB)
Microbiology
Models, Biological
oxidation
Oxygen
Oxygen - metabolism
Oxygen transfer
oxygen-limited fed batch (OLFB)
Pichia - enzymology
Pichia - genetics
Pichia - growth & development
Pichia pastoris
Protein Engineering - methods
Recombinant Proteins - biosynthesis
TECHNOLOGY
TEKNIKVETENSKAP
yeast
yield
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Summary: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.
ISSN:1615-7591
1615-7605
1615-7605
DOI:10.1007/s00449-005-0005-4