title: Efficient experimental design and micro-scale medium enhancement of 6-deoxyerythronolide B production through Escherichia coli.
abstract: The recent use of heterologous hosts to produce natural products has shown significant potential, although limitations still exist regarding optimal production titers. In this study, we utilize micro-scale cultures and well-defined screening methods to identify key medium components that influence the heterologous production of the complex polyketide 6-deoxyerythronolide B (6dEB) through E. coli. It was determined that tryptone had a significant effect on 6dEB production and could supplement substrate requirements and improve recombinant protein levels of the essential deoxyerythronolide B synthase (DEBS) which catalyze 6dEB conversion. As a result, the study (1) demonstrates the feasibility of micro-scale cultures to study E. coli 6dEB production and effectively model larger-scale cultures; (2) identifies an enhanced medium which generates over 160 mg L(-1) 6dEB (a 22-fold improvement over current culture media); and (3) provides new insight and understanding related to the heterologous production of 6dEB from E. coli.
authors.affiliation: Dept. of Chemical and Biological Engineering, Tufts University, Medford, MA 02155, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Pistorino
authors.forename: Blaine A
authors.initials: BA
authors.lastname: Pfeifer
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D000704:Analysis of Variance; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000379:methods*; D003470:Culture Media / Q000378:metabolism*; D003627:Data Interpretation, Statistical; D004917:Erythromycin / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D019216:Metals, Heavy / Q000378:metabolism; D010461:Peptones / Q000378:metabolism; D010865:Pilot Projects; D011422:Propionates / Q000378:metabolism; D015203:Reproducibility of Results
chemical_list: D000596:Amino Acids; D003470:Culture Media; D019216:Metals, Heavy; D010461:Peptones; D011422:Propionates; C034277:6-deoxyerythronolide B; D004917:Erythromycin; C001830:tryptones