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Strategies for high titre plasmid DNA production in Escherichia coli DH5α

Paper ID Volume ID Publish Year Pages File Format Full-Text
35857 45111 2007 11 PDF Available
Title
Strategies for high titre plasmid DNA production in Escherichia coli DH5α
Abstract

The production of plasmid pEGFP-N1 in Escherichia coli DH5α was optimised. A strategy evaluating different media components separately was not successful (OD < 2.5, low plasmid titres), a statistical approach via a Plackett Burman design (11 parameters) allowed some improvement (7 mg/L plasmid, OD600 8.5). Generally, high biomass did not correlate with high plasmid titres. When conditions were transferred to the bioreactor (batch operation) little improvement in plasmid titres (10 mg/L plasmid, OD600 20) was observed. By switching to a fed-batch procedure with linear feeding these values increased to 20 mg/L plasmid (OD600 50). By using an adaptive feeding strategy, plasmid titres could be increased to 50 mg/L. Finally, by combining a growth controlled (reduced temperature (35 °C), low dO2) initial batch phase with an adaptive feeding strategy in the fed-batch phase (37 °C, glucose-/dO2-limitation) we were reproducibly able to produce up to 250 mg/L of plasmid DNA in cultures that reached a final OD600 of 80.

Keywords
Escherichia coli; Fed batch; Media design; Nutrient limitation; Plackett Burman; Plasmid DNA
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 42, Issue 7, July 2007, Pages 1039–1049
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
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Any Questions? feel free to contact us