A recombinant lipoprotein antigen against Lyme disease expressed in E. coli: fermentor operating strategies for improved yield.

Printer-friendly versionPrinter-friendly versionPDF versionPDF version
TitleA recombinant lipoprotein antigen against Lyme disease expressed in E. coli: fermentor operating strategies for improved yield.
Publication TypeJournal Article
Year of Publication2000
AuthorsMadurawe, RD, Chase, TE, Tsao, EI, Bentley, WE
JournalBiotechnol Prog
Volume16
Issue4
Pagination571-6
Date Published2000 Jul-Aug
ISSN8756-7938
KeywordsAntigens, Bacterial, Bioreactors, Carbon, Culture Media, Escherichia coli, Fermentation, Glucose, Lipoproteins, Lyme Disease, Recombinant Proteins, Temperature
Abstract

Decorin-binding lipoprotein, lpp-DBP, a bacterial surface adhesin, shows promise as a vaccine against Lyme disease. It is expressed in recombinant E. coli as an undesirable 20.5 KDa apoprotein that is subsequently lipidated in vivo to the desired 22 KDa lpp-DBP form. This study defines fermentation conditions for maximizing lpp-DBP yield. Super broth medium, a low post-induction temperature (30 degrees C), and a glucose feed based on dissolved oxygen resulted in high lpp-DBP yield and minimized apoprotein formation. Since cells lysed within 2-3 h after induction, the cell yield was maximized by growing cells to high cell density prior to induction. Compared to a glucose feed based on maintaining a constant fermentor glucose concentration (Glucose-Stat), feeding based on maintaining a constant dissolved oxygen level (DO-Stat) improved yields. Also, a dissolved oxygen level of 60% (air saturation) was best, as no product degradation was detected by Western blotting and SDS-PAGE. Acetic acid levels under both modes of glucose feed were sufficiently low, and no adverse growth effects were observed.

DOI10.1021/bp0000555
Alternate JournalBiotechnol. Prog.
PubMed ID10933830