Metabolic engineering of Pseudomonas putida for methylmalonyl-CoA biosynthesis to enable complex heterologous secondary metabolite formation
Publikation: Beitrag in Fachzeitschrift › Forschungsartikel › Beigetragen › Begutachtung
Beitragende
Abstract
An operon consisting of three open reading frames, annotated in silico as methylmalonyl-CoA (mm-CoA) epimerase, mm-CoA mutase (MCM), and meaB, was identified in the sequencing project of the myxobacterium Sorangium cellulosum So ce56. This putative MCM pathway operon was subcloned from a bacterial artificial chromosome by Red/ET recombineering onto a minimal replicon derived from p15A. This plasmid was modified for integration and heterologous expression in Pseudomonas putida to enable the production of complex secondary metabolites requiring mm-CoA as precursor. Methylmalonate was identified in the recombinant P. putida strain by an analysis method based on gas chromatography/mass spectrometry. The engineered strain is able to synthesize polyketides requiring mm-CoA as an extender unit, which was demonstrated by the production of myxothiazol after integration of the biosynthetic gene cluster into the chromosome, followed by induction of expression.
Details
Originalsprache | Englisch |
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Seiten (von - bis) | 1253-64 |
Seitenumfang | 12 |
Fachzeitschrift | Chemistry and Biology |
Jahrgang | 13 |
Ausgabenummer | 12 |
Publikationsstatus | Veröffentlicht - Dez. 2006 |
Peer-Review-Status | Ja |
Externe IDs
PubMed | 17185221 |
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Scopus | 33845496065 |
ORCID | /0000-0002-4754-1707/work/142248089 |
Schlagworte
Schlagwörter
- Acyl Coenzyme A/biosynthesis, Amino Acid Sequence, Gene Expression Regulation, Gene Transfer Techniques, Genetic Engineering/methods, Methacrylates/metabolism, Molecular Sequence Data, Myxococcales/genetics, Operon/genetics, Pseudomonas putida/genetics, Sequence Alignment, Thiazoles/metabolism