論文 - 詳細
| RRC ID | 53635 |
|---|---|
| 著者 | Fang MY, Zhang C, Yang S, Cui JY, Jiang PX, Lou K, Wachi M, Xing XH. |
| タイトル | High crude violacein production from glucose by Escherichia coli engineered with interactive control of tryptophan pathway and violacein biosynthetic pathway. |
| ジャーナル | Microb Cell Fact |
| Abstract |
BACKGROUND:As bacteria-originated crude violacein, a natural indolocarbazole product, consists of violacein and deoxyviolacein, and can potentially be a new type of natural antibiotics, the reconstruction of an effective metabolic pathway for crude violacein (violacein and deoxyviolacein mixture) synthesis directly from glucose in Escherichia coli was of importance for developing industrial production process. RESULTS:Strains with a multivariate module for varied tryptophan productivities were firstly generated by combinatorial knockout of trpR/tnaA/pheA genes and overexpression of two key genes trpEfbr /trpD from the upstream tryptophan metabolic pathway. Then, the gene cluster of violacein biosynthetic pathway was introduced downstream of the generated tryptophan pathway. After combination of these two pathways, maximum crude violacein production directly from glucose by E. coli B2/pED+pVio was realized with a titer of 0.6±0.01 g L(-1) in flask culture, which was four fold higher than that of the control without the tryptophan pathway up-regulation. In a 5-L bioreactor batch fermentation with glucose as the carbon source, the recombinant E. coli B2/pED+pVio exhibited a crude violacein titer of 1.75 g L(-1) and a productivity of 36 mg L(-1) h(-1), which was the highest titer and productivity reported so far under the similar culture conditions without tryptophan addition. CONCLUSION:Metabolic pathway analysis using 13C labeling illustrated that the up-regulated tryptophan supply enhanced tryptophan metabolism from glucose, whereas the introduction of violacein pathway drew more carbon flux from glucose to tryptophan, thereby contributing to the effective production of crude violacein in the engineered E. coli cell factory. |
| 巻・号 | 14 |
| ページ | 8 |
| 公開日 | 2015-1-16 |
| DOI | 10.1186/s12934-015-0192-x |
| PII | s12934-015-0192-x |
| PMID | 25592762 |
| PMC | PMC4306242 |
| MeSH | Bacterial Proteins / genetics Bacterial Proteins / metabolism Biomass Carbon Isotopes / chemistry Carbon Isotopes / metabolism Corynebacterium / genetics Corynebacterium / metabolism Escherichia coli / genetics Escherichia coli / metabolism* Gene Knockout Techniques Glucose / metabolism* Indoles / metabolism* Metabolic Engineering* Multigene Family Plasmids / genetics Plasmids / metabolism Tryptophan / biosynthesis* |
| IF | 4.187 |
| 引用数 | 39 |
| リソース情報 | |
| 原核生物(大腸菌) | |