Overcoming glutamate auxotrophy in Escherichia coli itaconate overproducer by the Weimberg pathway.

Biosynthesis of itaconic acid occurs through decarboxylation of the TCA cycle intermediate cis-aconitate. Engineering of efficient itaconate producers often requires elimination of the highly active isocitrate dehydrogenase to conserve cis-aconitate, leading to 2-ketoglutarate auxotrophy in the producing strains. Supplementation of glutamate or complex protein hydrolysate then becomes necessary, often in large quantities, to support the high cell density desired during itaconate fermentation and adds to the production cost. Here, we present an alternative approach to overcome the glutamate auxotrophy in itaconate producers by synthetically introducing the Weimberg pathway from Burkholderia xenovorans for 2-ketoglutarate biosynthesis. Because of its independence from natural carbohydrate assimilation pathways in Escherichia coli, the Weimberg pathway is able to provide 2-ketoglutarate using xylose without compromising the carbon flux toward itaconate. With xylose concentration carefully tuned to minimize excess 2-ketoglutarate flux in the stationary phase, the final strain accumulated 20 g/L of itaconate in minimal medium from 18 g/L of xylose and 45 g/L of glycerol. Necessity of the recombinant Weimberg pathway for growth also allowed us to maintain multi-copy plasmids carrying in operon the itaconate-producing genes without addition of antibiotics. Use of the Weimberg pathway for growth restoration is applicable to other production systems with disrupted 2-ketoglutarate synthesis.

Dual-functional antibiofilm polymer composite for biodegradable medical devices.

Urinary tract infections (UTI) represent one of the most common problem within the urological disorders, and it is mainly caused by biofilm formation which leads to bacterial infection. Anti-adhesion and antibacterial agents are two primary mechanisms to prevent biofilm formation; however, current strategies are insufficiently effective. In this study, we developed an effective antibiofilm biodegradable polymer with high biocompatibility. Here we embedded silver nanoparticles (AgNPs) in poly(glycerol sebacate) acrylate (PGSA) followed by superhydrophilic modification on the polymer surfaces. The modified surfaces were characterized using SEM, AFM and contact angle measurements. This anti-adhesive surface prevented the adhesion of E. coli and limited the biofilm coverage percentage to less than 3% in 24 h. In the in vitro degradation, the long-term antibiofilm performance was evaluated in Nowatzki-Stoodley artificial urine (NSAU). The surface modified AgNPs embedded PGSA (sPGSA-AgNPs) was able to effectively inhibit the formation of biofilm by reducing the biofilm coverage to less than 0.01%, and it also showed low cytotoxicity with human bladder carcinoma cell. With the effective antibiofilm, biocompatibility and biodegradability, it is possible to be applied in urological devices to ameliorate the situation of UTIs.