Reference - Detail
| RRC ID | 35348 |
|---|---|
| Author | Boghigian BA, Zhang H, Pfeifer BA. |
| Title | Multi-factorial engineering of heterologous polyketide production in Escherichia coli reveals complex pathway interactions. |
| Journal | Biotechnol Bioeng |
| Abstract |
Polyketides represent a significant fraction of all natural products. Many possess pharmacological activity which makes them attractive drug candidates. The production of the parent macrocyclic aglycones is catalyzed by multi-modular polyketide synthases utilizing short-chain acyl-CoA monomers. When producing polyketides through heterologous hosts, one must not only functionally express the synthase itself, but activate the machinery used to generate the required substrate acyl-CoA's. As a result, metabolic engineering of these pathways is necessary for high-level production of heterologous polyketides. In this study, we over-express three different pathways for provision of the two substrates (propionyl-CoA and (2S)-methylmalonyl-CoA) utilized for the biosynthesis of 6-deoxyerythronolide B (6-dEB; the macrolactone precursor of erythromycin): (1) a propionate → propionyl-CoA → (2S)-methylmalonyl-CoA pathway, (2) a methylmalonate → methylmalonyl-CoA → propionyl-CoA pathway, and (3) a succinate → succinyl-CoA → (2R)-methylmalonyl-CoA → (2S)-methylmalonyl-CoA → propionyl-CoA pathway. The current study revealed that propionate is a necessary component for greater than 5 mg L(-1) titers. Deletion of the propionyl-CoA:succinate CoA transferase (ygfH) or over-expression of the transcriptional activator of short chain fatty acid uptake improved titer to over 100 mg L(-1), while the combination of the two improved titer to over 130 mg L(-1). The addition of exogenous methylmalonate could also improve titer to over 100 mg L(-1). Expression of a Streptomyces coelicolor A3(2) methylmalonyl-CoA epimerase, in conjunction with over-expression of Escherichia coli's native methylmalonyl-CoA mutase, allowed for the incorporation of exogenously fed succinate into the 6-dEB core. |
| Volume | 108(6) |
| Pages | 1360-71 |
| Published | 2011-6-1 |
| DOI | 10.1002/bit.23069 |
| PMID | 21337322 |
| PMC | PMC3076518 |
| MeSH | Base Sequence Erythromycin / analogs & derivatives* Erythromycin / metabolism Escherichia coli / genetics* Escherichia coli / metabolism Gene Expression Regulation, Bacterial* Macrolides / metabolism* Methylmalonic Acid / metabolism Methylmalonyl-CoA Mutase / genetics Methylmalonyl-CoA Mutase / metabolism Polyketide Synthases / genetics* Polyketide Synthases / metabolism Propionates / metabolism Signal Transduction |
| IF | 4.002 |
| Times Cited | 20 |
| WOS Category | BIOTECHNOLOGY & APPLIED MICROBIOLOGY |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 2 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Prokaryotes E. coli | JW2887-KC? |