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Introduction
This Application Report is part of a series documenting
culture growth in the BioFlo 110. With appropriate
vessels and control modules, the BioFlo 110 can efficiently
grow yeast and bacteria, as well as mammalian,
plant cells and insect cells.
Pichia pastoris:
Pichia pastoris is a methlotrophic yeast, which provides
a unique expression system for producing
high levels of recombinant protein, including
enzymes, proteases, protease inhibitors, receptors,
single-chain antibodies, and regulatory proteins at
various different levels. Pichia pastoris is also the
only system that offers the benefits of E. coli (cost
effective, high-level expression and easy scale-up)
combined with advantages of expression in a
eukaryotic system (protein processing, folding, and
posttranslational modifications).
A standard 7.5L BioFlo 110 Advanced Fermentation Kit was used to grow Pichia pastoris in a fed batch fermentation. We used BioCommand Plus supervisory software to control the feed schedule, achieving 91.0 g/L dry cell weight (DCW).
Next, a BioFlo 110 Gas-Mix Controller was added, and the fermentation repeated with oxygen supplementation of the sparge gas. This second run, described in the APPENDIX (page 5), achieved a very high dry cell weight of 177.4 g/L. Neither run was fully optimized, but the descriptions of procedures and materials, as well as the data discussion will be useful to operators of similar fermentors.
The Fermentor
Vessel
The BioFlo 110 Advanced Fermentation Kit, NBS
Catalog Number M1273-1125, was equipped with a
heat-blanketed 7.5 L fermentation vessel with nom
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Source:
Related biology technology :
1. High Yield Protein Production from Pichia pastoris Yeast: A Protocol for Benchtop Fermentation
2. E. coli Fermentation Using a BioFlo 110 Bench-Top Fermentor
3. Optimizing Your Benchscale Fermentation Process
4. Fermentation Cell Culture: University Style
5. Glossary for The BioPharm Guide to Fermentation and Cell Culture
6. Fundamentals of Fermentation: Techniques For Benchtop Fermentors
7. 140-Fold Increase of Protein Achieved in Pichia Fermentation
8. Outstanding transfection rates using the Multiporator
9. Outstanding transfection rates using the Multiporator
10. Isolation of Cosmids using Eppendorfs Perfectprep Plasmid Mini Kit
11. Normalizing cDNA libraries using the EppendorfThermomixer
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