High isoprenoid flux Escherichia coli as a host for carotenoids production

A noncarotenogenic microbe E. coli was engineered for high production of carotenoids. To increase the isoprenoid flux, the chromosomal native promoters of the rate-controlling steps (dxs, idi and ispDispF) in the isoprenoid pathway were replaced with a strong bacteriophage T5 promoter (P(T5)) by usi...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 834; p. 49
Main Author Suh, Wonchul
Format Journal Article
LanguageEnglish
Published United States 01.01.2012
Subjects
Carotenoids - biosynthesis
Chromosomes, Bacterial
Escherichia coli - genetics
Escherichia coli - metabolism
Gene Expression Regulation, Bacterial
Gene Order
Genetic Engineering - methods
Plasmids - genetics
Promoter Regions, Genetic
Recombination, Genetic
Terpenes - metabolism
Transduction, Genetic
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Summary:A noncarotenogenic microbe E. coli was engineered for high production of carotenoids. To increase the isoprenoid flux, the chromosomal native promoters of the rate-controlling steps (dxs, idi and ispDispF) in the isoprenoid pathway were replaced with a strong bacteriophage T5 promoter (P(T5)) by using the λ-Red recombinase system in combination with the Flp/FRT site-specific recombination system for marker excision and P1 transduction for gene trait stacking. The resulting high isoprenoid flux E. coli can be used as a starting strain to produce various carotenoids by introducing heterologous carotenoid genes. In this study, the high isoprenoid flux E. coli was transformed with a plasmid carrying the β-carotene biosynthetic genes from Pantoea stewartii for β-carotene production.
ISSN:1940-6029
DOI:10.1007/978-1-61779-483-4_4