Agitation rate effects on plasmid stability in immobilized and free-cell continuous cultures of recombinant E. coli.
Escherichia coli B/pTG201 recombinant cells were immobilized by entrapment in a carrageenan gel and cultivated in nonselective media to investigate the effect of agitation rate on plasmid stability, biomass concentration, and enzyme productivity. These parameters were studied in continuous cultures for free and immobilized cells, respectively. Immobilized recombinant cells exhibit an increase in the stability of the plasmid pTG201 compared to free cells, even under conditions where the tendency of plasmid stability for free cells decreased generally more rapidly under a higher agitation rate. Intensive agitation, resulting also in a strong shear stress, greatly reduced cell concentration within gel beads throughout the course of growth. Higher enzyme expression of catechol 2-3, dioxygenase was also obtained in leaked cells due to better maintenance of plasmid stability and higher plasmid copy number with regard to free cells. Enzyme productivity of leaked and free cells in minimal medium decreased with the increase in agitation rate, due to decreased plasmid stability; however, in LB medium, it increased in the presence of higher agitation rate related to important cell concentration.
Huang
J
J
Laboratoire de Technologie Enzymatique, Université de Technologie de Compiègne, France.
Dhulster
P
P
Thomas
D
D
Barbotin
J N
JN
D050557:Catechol 2,3-Dioxygenase; D049308:Dioxygenases; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development; D005818:Genetic Engineering / Q000379:methods*; D007700:Kinetics; D010105:Oxygenases / Q000096:biosynthesis / Q000235:genetics; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis
D011994:Recombinant Proteins; D010105:Oxygenases; D049308:Dioxygenases; D050557:Catechol 2,3-Dioxygenase