Debottlenecking recombinant protein production in Bacillus megaterium under large-scale conditions-targeted precursor feeding designed from metabolomics

2012 ◽  
Vol 109 (6) ◽  
pp. 1538-1550 ◽  
Author(s):  
Claudia Korneli ◽  
Christoph Josef Bolten ◽  
Thibault Godard ◽  
Ezequiel Franco-Lara ◽  
Christoph Wittmann
Keyword(s):  
Recombinant Protein ◽  
Bacillus Megaterium ◽  
Recombinant Protein Production ◽  
Large Scale ◽  
Protein Production ◽  
Precursor Feeding

scholarly journals Polar Fixation of Plasmids during Recombinant Protein Production in Bacillus megaterium Results in Population Heterogeneity

2015 ◽  
Vol 81 (17) ◽  
pp. 5976-5986 ◽  
Author(s):  
Karin M. Münch ◽  
Johannes Müller ◽  
Sarah Wienecke ◽  
Simone Bergmann ◽  
Steffi Heyber ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Division ◽  
Recombinant Protein ◽  
Bacillus Megaterium ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Population Heterogeneity ◽  
Exponential Growth Phase ◽  
Scale Production ◽  
Content Type

ABSTRACTDuring the past 2 decades,Bacillus megateriumhas been systematically developed for the gram-per-liter scale production of recombinant proteins. The plasmid-based expression systems employed use a xylose-controlled promoter. Protein production analyses at the single-cell level using green fluorescent protein as a model product revealed cell culture heterogeneity characterized by a significant proportion of less productive bacteria. Due to the enormous size ofB. megaterium, such bistable behavior seen in subpopulations was readily analyzed by time lapse microscopy and flow cytometry. Cell culture heterogeneity was not caused simply by plasmid loss: instead, an asymmetric distribution of plasmids during cell division was detected during the exponential-growth phase. Multicopy plasmids are generally randomly distributed between daughter cells. However,in vivo andin vitroexperiments demonstrated that under conditions of strong protein production, plasmids are retained at one of the cell poles. Furthermore, it was found that cells with accumulated plasmids and high protein production ceased cell division. As a consequence, the overall protein production of the culture was achieved mainly by the subpopulation with a sufficient plasmid copy number. Based on our experimental data, we propose a model whereby the distribution of multicopy plasmids is controlled by polar fixation under protein production conditions. Thereby, cell lines with fluctuating plasmid abundance arise, which results in population heterogeneity. Our results provide initial insights into the mechanism of cellular heterogeneity during plasmid-based recombinant protein production in aBacillusspecies.

Systems Biology of Recombinant Protein Production in Bacillus megaterium

2010 ◽  
pp. 133-161 ◽  
Author(s):  
Rebekka Biedendieck ◽  
Boyke Bunk ◽  
Tobias Fürch ◽  
Ezequiel Franco-Lara ◽  
Martina Jahn ◽  
...  
Keyword(s):  
Systems Biology ◽  
Recombinant Protein ◽  
Bacillus Megaterium ◽  
Recombinant Protein Production ◽  
Protein Production

Bacillus megaterium as a Host for Recombinant Protein Production

2006 ◽  
Vol 6 (5) ◽  
pp. 470-474
Author(s):  
R. Hollmann ◽  
M. Malten ◽  
R. Biedendieck ◽  
Y. Yang ◽  
W. Wang ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Bacillus Megaterium ◽  
Recombinant Protein Production ◽  
Protein Production

scholarly journals Large-scale recombinant protein production and structure-based drug design capabilities at CSIRO

2010 ◽  
Vol 31 (2) ◽  
pp. 77
Author(s):  
G Lovrecz ◽  
G Dumsday ◽  
T Adams
Keyword(s):  
Drug Design ◽  
Recombinant Protein ◽  
Collaborative Research ◽  
Recombinant Protein Production ◽  
Large Scale ◽  
Protein Production ◽  
Research Infrastructure ◽  
Structure Based Drug Design ◽  
Production Facilities

CSIRO has opened its large-scale Recombinant Protein Production Facilities (RPPF) as part of the National Collaborative Research Infrastructure Scheme (NCRIS).

Large-Scale Insect Cell-Culture for Recombinant Protein Production

1988 ◽  
Vol 6 (12) ◽  
pp. 1406-1410 ◽  
Author(s):  
Brian Maiorella ◽  
Duane Inlow ◽  
Andrea Shauger ◽  
David Harano
Keyword(s):  
Cell Culture ◽  
Recombinant Protein ◽  
Insect Cell ◽  
Recombinant Protein Production ◽  
Large Scale ◽  
Protein Production ◽  
Scale Insect ◽  
Insect Cell Culture
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