Identification of isopentenol biosynthetic genes from Bacillus subtilis by a screening method based on isoprenoid precursor toxicity.
Appl Environ Microbiol
; 73(19): 6277-83, 2007 Oct.
Article
en En
| MEDLINE
| ID: mdl-17693564
ABSTRACT
We have developed a novel method to clone terpene synthase genes. This method relies on the inherent toxicity of the prenyl diphosphate precursors to terpenes, which resulted in a reduced-growth phenotype. When these precursors were consumed by a terpene synthase, normal growth was restored. We have demonstrated that this method is capable of enriching a population of engineered Escherichia coli for those clones that express the sesquiterpene-producing amorphadiene synthase. In addition, we enriched a library of genomic DNA from the isoprene-producing bacterium Bacillus subtilis strain 6,051 in E. coli engineered to produce elevated levels of isopentenyl diphosphate and dimethylallyl diphosphate. The selection resulted in the discovery of two genes (yhfR and nudF) whose protein products acted directly on the prenyl diphosphate precursors and produced isopentenol. Expression of nudF in E. coli engineered with the mevalonate-based isopentenyl pyrophosphate biosynthetic pathway resulted in the production of isopentenol.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Pentanos
/
Terpenos
/
Bacillus subtilis
/
Butadienos
/
Ingeniería Genética
/
Transferasas Alquil y Aril
/
Hemiterpenos
/
Pentanoles
Tipo de estudio:
Diagnostic_studies
/
Screening_studies
Idioma:
En
Revista:
Appl Environ Microbiol
Año:
2007
Tipo del documento:
Article
País de afiliación:
Estados Unidos