High cell density Escherichia coli cultivation in a BIOSTAT®A - 4 Pages

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High cell density Escherichia coli cultivation in a BIOSTAT®A

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sartoriusstedim biotech High cell density Escherichia coli cultivation in a BIOSTAT® A turning science

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Thomas Dreher, Matthias Muller, Ute Husemann, Gerhard Greller, Sonke Rosemann Sartorius Stedim Biotech Introduction Escherichia coli is commonly used as a host organism for recombinant protein production. Due to its ease of handling, well known metabolism and genetic characterization, Escherichia coli strains like BL21 (DE3) are used for training purposes especially in educational environments. Moreover, many protocols for batch as well as fed-batch cultivations of Escherichia coli strains are available. Fermentation workshops should focus on key learning aspects of controlled cell growth...

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2. Results As a case study, a high cell density Escherichia coli fermentation was performed to assess if a BIOSTAT® A fulfills the requirements of this challenging process. In figure 3 the characteristics of the OD600 (proportional to the cell density increase) and the specific growth rate are shown to verify the growth behavior. Figure 1: BIOSTAT® A setup for the Escherichia coli high cell density fermentation experiment Once the glucose in the culture medium was completely consumed a highly concentrated feed was supplied with an exponential increasing rate. The specific growth rate was...

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Figure 4 shows the Dissolved Oxygen, temperature and pH measured value profiles. These may be used to evaluate if the culture conditions were optimal. Within the first 6 hours the pO2 decreased exponentially caused by increasing oxygen uptake rate until the set point of 20% was reached. After t = 7.5 h a significant pO2 increase was observed, indicating the complete consumption of the initial glucose and the end of the batch phase. During the remainder of the fermentation the pO2 was adequately controlled to the set point. Hence, it can be concluded that aerobic conditions were present for...

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