( RNA molecules we require multipleki...)Fermentation Cell Culture: University Style

RNA molecules, we require multiple kilograms of yeast. The strains we use for these experiments contain specific modifications in several genes to facilitate the snRNP purifications, so we are unable to use commercially available wild-type yeast cakes.

BioPharm: What cultures do you grow in the laboratory?

Stevens: We primarily grow Saccharomyces cerevisiae in our laboratory, but we also grow Schizosaccharomyces pombe and Escherichia coli.

BioPharm: For what purpose(s) do you grow the cultures?

Stevens: For our large-scale microorganism growth, we generally use the product for protein or macromolecular complex purification. For S. cerevisiae and E. coli we perform preparative purifications of single polypeptides ( E. coli ), or multi-protein complexes ( S. cerevisiae ). For S. pombe we perform exploratory analyses of macromolecular complex composition.

BioPharm: Describe the processes and technology you originally employed to grow cultures and the progression to those you use today?

Stevens: Like most academic investigators performing similar experiments, we began experimentation using small-scale cultures of one to six liters in shake-flasks. We continue to perform small-scale experiments to troubleshoot and optimize our purification procedures. We now are able to routinely produce 400 liters of culture in a fermentor (500L modular BioFlo Pro, New Brunswick Scientific [NBS]) in an overnight experiment. We find that
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