Maximizing filamentous phage yield during computer-controlled fermentation.
Filamentous phage such as M13 and fd consist of a circular, single-stranded DNA molecule surrounded by several different coat proteins. These phages have been used extensively as vectors in phage display where one of the phage coat proteins is genetically engineered to contain a unique peptide surface loop. Through these peptide sequences, a phage collection can be screened for individual phage that binds to different macromolecules or small organic and inorganic molecules. Here, we use computer-controlled bioreactors to produce large quantities of filamentous phage in the bacterial host Escherichia coli. By measuring phage yield and bacterial growth while changing the growth medium, pH and dissolved oxygen concentration, we found that the optimal conditions for phage yield were NZY medium with pH maintained at 7.4, the dO(2) held at 100% and agitation at 800 rpm. These computer-controlled fermentations result in a minimum of a tenfold higher filamentous phage production compared to standard shake flask conditions.
Grieco
Sung-Hye H
SH
Centre for Blood Research, University of British Columbia, Vancouver, Canada.
Lee
Seungil
S
Dunbar
W Scott
WS
MacGillivray
Ross T A
RT
Curtis
Susan B
SB
D001698:Biomedical Engineering; D019149:Bioreactors / Q000382:microbiology / Q000821:virology*; D003201:Computers; D003470:Culture Media; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000821:virology; D005285:Fermentation; D005818:Genetic Engineering; D006863:Hydrogen-Ion Concentration; D017912:Inovirus / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology*; D007700:Kinetics; D019151:Peptide Library; D014773:Virology / Q000379:methods*
D003470:Culture Media; D019151:Peptide Library