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Once a clone has been selected that expresses a sufficient amount of soluble, active serpin, glycerol stocks of that clone should be prepared and frozen at )80 C toensure a continuous supply of viable yeast. Glycerol stocks can be used to grow starter cultures that in turn can be used to seed larger cultures for yeast growth and expression of the serpin. The needs of most academic research are satisfied by growth and expression at the 1–5 L scale in shake flasks or small fermentors, allowing the recovery and purification of milligram to gram quantities of product, but development of a commercial fermentation process requires considerable additional work to optimize the metabolic requirements of that yeast clone to maximize biomass and serpin expression. A few of these are: batch versus continuous culture (primarily for secreted serpins), selection of optimal growth media, amino acid feed composition, feed rate, dissolved oxygen (DO2) delivery, and aeration rate.
Once a clone has been selected that expresses a sufficient amount of soluble, active serpin, glycerol stocks of that clone should be prepared and frozen at )80 C toensure a continuous supply of viable yeast. Glycerol stocks can be used to grow starter cultures that in turn can be used to seed larger cultures for yeast growth and expression of the serpin. The needs of most academic research are satisfied by growth and expression at the 1–5 L scale in shake flasks or small fermentors, allowing the recovery and purification of milligram to gram quantities of product, but development of a commercial fermentation process requires considerable additional work to optimize the metabolic requirements of that yeast clone to maximize biomass and serpin expression. A few of these are: batch versus continuous culture (primarily for secreted serpins), selection of optimal growth media, amino acid feed composition, feed rate, dissolved oxygen (DO2) delivery, and aeration rate.
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