Optimization of fermentation parameters in phage production using response surface methodology.
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.
Centre for Blood Research, University of British Columbia, Vancouver, BC V6T 1Z3, Canada. sunghye.grieco@ubc.ca
Sung-Hye H
SH
Grieco
Ann Y K
AY
Wong
W Scott
WS
Dunbar
Ross T A
RT
MacGillivray
Susan B
SB
Curtis
D000704:Analysis of Variance; D004926:Escherichia coli / Q000378:metabolism / Q000821:virology*; D005285:Fermentation; D006863:Hydrogen-Ion Concentration; D017912:Inovirus / Q000254:growth & development*; D008954:Models, Biological; D010100:Oxygen / Q000378:metabolism; D015203:Reproducibility of Results; D013696:Temperature
D010100:Oxygen