Controlling catabolite repression for isobutanol production using glucose and xylose by overexpressing the xylose regulator.

Using lignocellulosic biomass is immensely beneficial for the economical production of biochemicals. However, utilizing mixed sugars from lignocellulosic biomass is challenging because of bacterial preference for specific sugar such as glucose. Although previous studies have attempted to overcome this challenge, no studies have been reported on isobutanol production from mixed sugars in the Escherichia coli strain. To overcome catabolite repression of xylose and produce isobutanol using mixed sugars, we applied the combination of three strategies: (1) deletion of the gene for the glucose-specific transporter of the phosphotransferase system (ptsG); (2) overexpression of glucose kinase (glk) and glucose facilitator protein (glf); and (3) overexpression of the xylose regulator (xylR). xylR gene overexpression resulted in 100% of glucose and 82.5% of xylose consumption in the glucose-xylose mixture (1:1). Moreover, isobutanol production increased by 192% in the 1:1 medium, equivalent to the amount of isobutanol produced using only glucose. These results indicate the effectiveness of xylR overexpression in isobutanol production. Our findings demonstrated various strategies to overcome catabolite repression for a specific product, isobutanol. The present study suggests that the selected strategy in E. coli could overcome the major challenge using lignocellulosic biomass to produce isobutanol.

Coproduction of exopolysaccharide and polyhydroxyalkanoates from Sphingobium yanoikuyae BBL01 using biochar pretreated plant biomass hydrolysate.

Sphingobium yanoikuyae BBL01 can produce exopolysaccharides (EPS) and polyhydroxyalkanoates (PHAs). The effect of side products (furfural, hydroxymethylfurfural (HMF), vanillin, and acetate) produced during pretreatment of biomass was evaluated on S. yanoikuyae BBL01. It was observed that a certain concentration range (0.01-0.03 %) of these compounds can improve growth, EPS production, and polyhydroxybutyrate (PHB) accumulation. The addition of HMF increases glucose and xylose utilization while other side products have a negative effect. The C/N of 5 favors EPS production (3.24 ± 0.05 g/L), while a higher C/N ratio of 30 promotes PHB accumulation (38.7 ± 0.08 % w/w), when commercial sugar is used as a carbon source. Pine biomass-derived biochar was able to remove 40 ± 2.1 % of total phenolic. Various biomass hydrolysates were evaluated and the use of detoxified pine biomass hydrolysate (DPH) as a carbon source resulted in the higher coproduction of EPS (2.83 ± 0.03 g/L) and PHB (40.8 ± 2.4 % w/w).

Leucyl-tRNA Synthetase Inhibitor, D-Norvaline, in Combination with Oxacillin, Is Effective against Methicillin-Resistant Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) is a pathogenic bacterium that causes severe diseases in humans. For decades, MRSA has acquired substantial resistance against conventional antibiotics through regulatory adaptation, thereby posing a challenge for treating MRSA infection. One of the emerging strategies to combat MRSA is the combinatory use of antibacterial agents. Based on the dramatic change in phospholipid fatty acid (PLFA) composition of MRSA in previous results, this study investigated branched-chain amino acid derivatives (precursors of fatty acid synthesis of cell membrane) and discovered the antimicrobial potency of D-norvaline. The compound, which can act synergistically with oxacillin, is among the three leucine-tRNA synthetase inhibitors with high potency to inhibit MRSA cell growth and biofilm formation. PLFA analysis and membrane properties revealed that D-norvaline decreased the overall amount of PLFA, increasing the fluidity and decreasing the hydrophobicity of the bacterial cell membrane. Additionally, we observed genetic differences to explore the response to D-norvaline. Furthermore, deletion mutants and clinically isolated MRSA strains were treated with D-norvaline. The study revealed that D-norvaline, with low concentrations of oxacillin, was effective in killing several MRSA strains. In summary, our findings provide a new combination of aminoacyl-tRNA synthetase inhibitor D-norvaline and oxacillin, which is effective against MRSA.