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 |
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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 |
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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