Influence of controlled glucose oscillations on a fed-batch process of recombinant Escherichia coli.
The influence of glucose oscillations on cell growth and product formation of a recombinant Escherichia coli culture producing a heterologous alpha-glucosidase was studied in fed-batch cultures in a laboratory bioreactor. Glucose oscillations were created by an on/off-feeding mode in either fast cycles (1 min) or slow cycles (4 min) and compared to a process with constant glucose addition. The study indicates that glucose oscillations influence the product stability and the overgrowth of plasmid-free cells if such cultures are not performed under continuous pressure for selection of plasmid-containing cells. Although the glucose uptake capacity decreased after induction of the recombinant alpha-glucosidase in all cultures performed, the up-growth of plasmid-free cells during the production phase was strongly inhibited by fast oscillations. In contrast, plasmid-free cells grew up when constant feeding or slow cycles were applied. Our data suggest that the various feed protocols effect the specific carbon dioxide formation rate differently, with the highest production of carbon dioxide in the cultivations with fast cycles. In connection to product formation the initial alpha-glucosidase accumulation was the same in all cultures, but the stability of the product was significantly lower in the cultivation with slow cycles. Our results from laboratory experiments are discussed in relation to the mixing situation in large-scale bioreactors.
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Martin-Luther-Universität Halle-Wittenberg, Fachbereich Biochemie /Biotechnologie, Institut für Biotechnologie, Halle, Germany.
Neubauer
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D019149:Bioreactors; D015169:Colony Count, Microbial; D004274:DNA, Recombinant; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development*; D005285:Fermentation; D005947:Glucose / Q000378:metabolism*; D007700:Kinetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D000520:alpha-Glucosidases / Q000096:biosynthesis
D004274:DNA, Recombinant; D011994:Recombinant Proteins; D000520:alpha-Glucosidases; D005947:Glucose