title: Increased bacterial cell density and recombinant protein yield using a commercial microbial cultivation system.
abstract: EnBase (BioSilta, Finland) is a microbial cultivation system that replicates fed-batch systems through sustained release of glucose by enzymatic degradation of a polymeric substrate. Achievable bacterial cell densities and recombinant capripoxvirus protein expression levels, solubility, and antigenicity using the EnBase system were assessed. BL21-AI Escherichia coli expressing capripoxvirus proteins achieved up to eightfold higher cell densities when grown in EnBase media compared with standard media. Greater yields of capripoxvirus proteins were attained using EnBase media, either through increases in the amount of expressed protein per cell in conjunction with higher cell density or through the increase in cell density alone. Addition of EnBase booster enhanced protein yield for one of the proteins tested but reduced yield for the other. However, the amount of soluble forms of the capripoxvirus proteins tested was not different from that observed from cultures grown under standard conditions. Purified capripoxvirus proteins expressed using EnBase or standard media were assessed for their performance by enzyme-linked immunosorbent assay (ELISA) and were shown to be equally capable of specifically binding capripoxvirus antibodies.
authors.affiliation: a CSIRO Animal, Food and Health Sciences, Australian Animal Health Laboratory , East Geelong , Victoria , Australia.
authors.forename: Grantley R
authors.initials: GR
authors.lastname: Peck
authors.forename: Timothy R
authors.initials: TR
authors.lastname: Bowden
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Shiell
authors.forename: Wojtek P
authors.initials: WP
authors.lastname: Michalski
mesh_terms: D019149:Bioreactors; D018151:Capripoxvirus / Q000235:genetics*; D003001:Cloning, Molecular; D003470:Culture Media / Q000378:metabolism; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics* / Q000378:metabolism; D007218:Industrial Microbiology / Q000295:instrumentation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D012995:Solubility; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D003470:Culture Media; D011994:Recombinant Proteins; D014764:Viral Proteins