Optimization of fermentation parameters in phage production using response surface methodology |
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Authors: | Sung-Hye H. Grieco Ann Y. K. Wong W. Scott Dunbar Ross T. A. MacGillivray Susan B. Curtis |
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Affiliation: | Centre for Blood Research, University of British Columbia, 2350 Health Sciences Mall, Vancouver, BC, V6T 1Z3, Canada, sunghye.grieco@ubc.ca. |
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Abstract: | ![]() Previously, we used computer-controlled fermentation technology to improve the yield of filamentous phage produced in Escherichia coli by 10-fold (Grieco et?al., Bioprocess Biosyst Eng 32:773-779, 2009). In the current study, three major fermentation parameters (temperature, dissolved oxygen [DO], and pH) were investigated using design of experiments (DOE) methodology. Response surface methodology (RSM) was employed to create a process model and determine the optimal conditions for maximal phage production. The experimental data fitted best to a quadratic model (p?0.0001). Temperature and pH, but not DO, proved to be significant variables. The model predicted a theoretical optimal condition for maximal bacteriophage production at temperature of 28.1?°C and pH 6.9. A validation run resulted in phage production [3.49?×?10(11)?transducing units (TU)/mL] comparable to the predicted value (2.86?×?10(11)?TU/mL). This represented a 7-fold increase in phage production above that obtained without optimization, resulting in a 70-fold increase above that achieved by shake flask culture alone. |
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