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Efficient experimental design and micro-scale medium enhancement of 6-deoxyerythronolide B production through Escherichia coli.

Biotechnology progress (2009-07-28)
Michael Pistorino, Blaine A Pfeifer
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Peptone from animal tissue, from meat, BioReagent, suitable for cell culture, suitable for plant cell culture
Millipore
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Peptone from casein and other animal proteins, suitable for microbiology
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Peptone Primatone® RL, suitable for microbiology
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Peptone Primatone® RLT, suitable for microbiology
Sigma-Aldrich
Peptone from animal tissue, Type I, BioReagent, suitable for plant cell culture, from meat