Lactose autoinduction with enzymatic glucose release: characterization of the cultivation system in bioreactor.
The lactose autoinduction system for recombinant protein production was combined with enzymatic glucose release as a method to provide a constant feed of glucose instead of using glycerol as a carbon substrate. Bioreactor cultivation confirmed that the slow glucose feed does not prevent the induction by lactose. HPLC studies showed that with successful recombinant protein production only a very low amount of lactose was metabolized during glucose-limited fed-batch conditions by the Escherichia coli strain BL21(DE3)pLysS in well-aerated conditions, which are problematic for glycerol-based autoinduction systems. We propose that slow enzymatic glucose feed does not cause a full activation of the lactose operon. However recombinant PDI-A protein (A-domain of human disulfide isomerase) was steadily produced until the end of the cultivation. The results of the cultivations confirmed our earlier observations with shaken cultures showing that lactose autoinduction cultures based on enzymatic glucose feed have good scalability, and that this system can be applied also to bioreactor cultivations.
BioSilta Oy, P.O. Box 4300, FI-90014 Oulu, Finland; Chair of Bioprocess Engineering, Department of Biotechnology, Technische Universität Berlin, Ackerstrasse 76, ACK24, D-13355 Berlin, Germany. Electronic address: sonja.mayer@uni-tuebingen.de.
Sonja
S
Mayer
Chair of Bioprocess Engineering, Department of Biotechnology, Technische Universität Berlin, Ackerstrasse 76, ACK24, D-13355 Berlin, Germany. Electronic address: stefan.junne@tu-berlin.de.
Stefan
S
Junne
BioSilta Oy, P.O. Box 4300, FI-90014 Oulu, Finland; Bioprocess Engineering Laboratory, Department of Process and Environmental Engineering, University of Oulu, P.O. Box 4300, FI-90014 Oulu, Finland. Electronic address: kaisa.ukkonen@oulu.fi.
Kaisa
K
Ukkonen
Chair of Bioprocess Engineering, Department of Biotechnology, Technische Universität Berlin, Ackerstrasse 76, ACK24, D-13355 Berlin, Germany. Electronic address: julia.glazyrina@tu-berlin.de.
Julia
J
Glazyrina
Chair of Bioprocess Engineering, Department of Biotechnology, Technische Universität Berlin, Ackerstrasse 76, ACK24, D-13355 Berlin, Germany. Electronic address: florian.glauche@tu-berlin.de.
Florian
F
Glauche
Chair of Bioprocess Engineering, Department of Biotechnology, Technische Universität Berlin, Ackerstrasse 76, ACK24, D-13355 Berlin, Germany. Electronic address: peter.neubauer@tu-berlin.de.
Peter
P
Neubauer
BioSilta Oy, P.O. Box 4300, FI-90014 Oulu, Finland. Electronic address: antti.vasala@biosilta.com.
Antti
A
Vasala
D019149:Bioreactors; D003470:Culture Media / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005947:Glucose / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007785:Lactose / Q000378:metabolism* / Q000494:pharmacology; D011393:Procollagen-Proline Dioxygenase / Q000096:biosynthesis / Q000235:genetics; D019704:Protein Disulfide-Isomerases / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics
D003470:Culture Media; D011994:Recombinant Proteins; D011393:Procollagen-Proline Dioxygenase; C526497:P4HB protein, human; D019704:Protein Disulfide-Isomerases; D005947:Glucose; D007785:Lactose
Bioreactor; Enzymatic glucose delivery; Fed-batch; Lactose autoinduction