Metabolic engineering of Pseudomonas putida for methylmalonyl-CoA biosynthesis to enable complex heterologous secondary metabolite formation
Research output: Contribution to journal › Research article › Contributed › peer-review
Contributors
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
| Original language | English |
|---|---|
| Pages (from-to) | 1253-64 |
| Number of pages | 12 |
| Journal | Chemistry and Biology |
| Volume | 13 |
| Issue number | 12 |
| Publication status | Published - Dec 2006 |
| Peer-reviewed | Yes |
External IDs
| PubMed | 17185221 |
|---|---|
| Scopus | 33845496065 |
| ORCID | /0000-0002-4754-1707/work/142248089 |
Keywords
Keywords
- 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