In silico and in vivo stability analysis of a heterologous biosynthetic pathway for 1,4-butanediol production in metabolically engineered E. coli
| Autor: | Szabolcs Lányi, Beáta Albert, Zsolt Bodor, Réka Sinkler, Kálmán Csongor Orbán, Ildikó Miklóssy |
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| Rok vydání: | 2016 |
| Předmět: |
Glycerol
0301 basic medicine In silico Heterologous Biology Xylose medicine.disease_cause Metabolic engineering Gene Knockout Techniques 03 medical and health sciences chemistry.chemical_compound Biosynthesis Structural Biology Escherichia coli medicine Computer Simulation Butylene Glycols Molecular Biology General Medicine Biosynthetic Pathways Flux balance analysis Bacteria Aerobic Glucose 030104 developmental biology Metabolic Engineering chemistry Biochemistry Fermentation |
| DOI: | 10.6084/m9.figshare.3531320.v1 |
| Popis: | Recently, several approaches have been published in order to develop a functional biosynthesis route for the non-natural compound 1,4-butanediol (BDO) in E. coli using glucose as a sole carbon source or starting from xylose. Among these studies, there was reported as high as 18 g/L product concentration achieved by industrial strains, however BDO production varies greatly in case of the reviewed studies. Our motivation was to build a simple heterologous pathway for this compound in E. coli and to design an appropriate cellular chassis based on a systemic biology approach, using constraint-based flux balance analysis and bi-level optimization for gene knock-out prediction. Thus, the present study reports, at the “proof-of concept” level, our findings related to model-driven development of a metabolically engineered E. coli strain lacking key genes for ethanol, lactate and formate production (ΔpflB, ΔldhA and ΔadhE), with a three-step biosynthetic pathway. We found this strain to produce a limited quantity of 1,4-BDO (.89 mg/L BDO under microaerobic conditions and .82 mg/L under anaerobic conditions). Using glycerol as carbon source, an approach, which to our knowledge has not been tackled before, our results suggest that further metabolic optimization is needed (gene-introductions or knock-outs, promoter fine-tuning) to address the redox potential imbalance problem and to achieve development of an industrially sustainable strain. Our experimental data on culture conditions, growth dynamics and fermentation parameters can consist a base for ongoing research on gene expression profiles and genetic stability of such metabolically engineered E. coli strains. |
| Databáze: | OpenAIRE |
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