No abstract

No abstract

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.

Succinic acid production through biological fermentation led to new pathways in the integration of renewable feedstock from different industries into biosynthesis. In this article, we investigate the population growth dynamics and succinic acid production potential of the recently isolated natural succinic acid producer, Basfia succiniciproducens, using in silico constraint-based metabolic models as well as in vitro experiments. Our work focuses on the influence of different renewable substrates and added yeast extract on fermentation dynamics, and the produced metabolites of the strain cultured in mineral (minimal) medium. According to our experiments, which were carried out as small-scale fermentations and in bioreactor conditions, glucose is the preferred carbon source, while the addition of 1% yeast extract has a significant positive effect on biomass formation. In the case of B. succiniciproducens cultured in minimal salt medium, a production potential as high as 47.09 mM succinic acid was obtained in these conditions. Industrial applications related to this bacterial strain could contribute to new possibilities for the re-use of byproducts by using fermentation processes, leading to high added-value compounds.

No abstract