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P. pastoris Fermentation using a BioFlo 110 Benchtop Fermentor


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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