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- Title
Efficient experimental design and micro-scale medium enhancement of 6-deoxyerythronolide B production through Escherichia coli.
- Authors
Pistorino, Michael; Pfeifer, Blaine A.
- 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. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009
- Subjects
EXPERIMENTAL design; ESCHERICHIA coli; CELL culture; POLYKETIDES; POLYMERS; CULTURE media (Biology)
- Publication
Biotechnology Progress, 2009, Vol 25, Issue 5, p1364
- ISSN
8756-7938
- Publication type
Article
- DOI
10.1002/btpr.250