Systems biology of recombinant protein production in Bacillus megaterium.
Over the last two decades the Gram-positive bacterium Bacillus megaterium was systematically developed to a useful alternative protein production host. Multiple vector systems for high yield intra- and extracellular protein production were constructed. Strong inducible promoters were combined with DNA sequences for optimised ribosome binding sites, various leader peptides for protein export and N- as well as C-terminal affinity tags for affinity chromatographic purification of the desired protein. High cell density cultivation and recombinant protein production were successfully tested. For further system biology based control and optimisation of the production process the genomes of two B. megaterium strains were completely elucidated, DNA arrays designed, proteome, fluxome and metabolome analyses performed and all data integrated using the bioinformatics platform MEGABAC. Now, solid theoretical and experimental bases for primary modeling attempts of the production process are available.
Protein Science Group, Department of Biosciences, University of Kent, Canterbury, Kent, CT27NJ, UK.
Rebekka
R
Biedendieck
Boyke
B
Bunk
Tobias
T
Fürch
Ezequiel
E
Franco-Lara
Martina
M
Jahn
Dieter
D
Jahn
D001410:Bacillus megaterium / Q000502:physiology*; D001426:Bacterial Proteins / Q000502:physiology*; D003198:Computer Simulation; D024861:Genetic Enhancement / Q000379:methods*; D008957:Models, Genetic; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D049490:Systems Biology / Q000379:methods*
D001426:Bacterial Proteins; D011994:Recombinant Proteins