FAST, a method based on split-GFP for the detection in solution of proteins synthesized in cell-free expression systems.

Cell-free protein synthesis (CFPS) systems offer a versatile platform for a wide range of applications. However, the traditional methods for detecting proteins synthesized in CFPS, such as radioactive labeling, fluorescent tagging, or electrophoretic separation, may be impractical, due to environmental hazards, high costs, technical complexity, and time consuming procedures. These limitations underscore the need for new approaches that streamline the detection process, facilitating broader application of CFPS. By harnessing the reassembly capabilities of two GFP fragments-specifically, the GFP1-10 and GFP11 fragments-we have crafted a method that simplifies the detection of in vitro synthesized proteins called FAST (Fluorescent Assembly of Split-GFP for Translation Tests). FAST relies on the fusion of the small tag GFP11 to virtually any gene to be expressed in CFPS. The in vitro synthesized protein:GFP11 can be rapidly detected in solution upon interaction with an enhanced GFP1-10 fused to the Maltose Binding Protein (MBP:GFP1-10). This interaction produces a fluorescent signal detectable with standard fluorescence readers, thereby indicating successful protein synthesis. Furthermore, if required, detection can be coupled with the purification of the fluorescent complex using standardized MBP affinity chromatography. The method's versatility was demonstrated by fusing GFP11 to four distinct E. coli genes and analyzing the resulting protein synthesis in both a homemade and a commercial E. coli CFPS system. Our experiments confirmed that the FAST method offers a direct correlation between the fluorescent signal and the amount of synthesized protein:GFP11 fusion, achieving a sensitivity threshold of 8 ± 2 pmol of polypeptide, with fluorescence plateauing after 4 h. Additionally, FAST enables the investigation of translation inhibition by antibiotics in a dose-dependent manner. In conclusion, FAST is a new method that permits the rapid, efficient, and non-hazardous detection of protein synthesized within CFPS systems and, at the same time, the purification of the target protein.

Practical High-Voltage Lithium Metal Batteries Enabled by Tuning the Solvation Structure in Weakly Solvating Electrolyte.

Li metal batteries (LMBs) are ideal candidates for future high-energy-density battery systems. To date, high-voltage LMBs suffer severe limitations because of electrolytes unstable against Li anodes and high-voltage cathodes. Although ether-based electrolytes exhibit good stability with Li metal, compared to carbonate-based electrolytes, they have been used only in ≤4.0 V LMBs because of their limited oxidation stability. Here, a high concentration electrolyte (HCE) comprising lithium bis(fluorosulfonyl)imide (LiFSI) and a weakly solvating solvent (1,2-diethoxyethane, DEE) is designed, which can regulate unique solvation structures with only associated complexes at relatively lower concentration compared to the reported HCEs. This effectively suppresses dendrites on the anode side, and preserves the structural integrity of the cathode side under high voltages by the formation of stable interfacial layers on a Li metal anode and LiNi0.8 Mn0.1 Co0.1 O2 (NMC811) cathode. Consequently, a 3.5 m LiFSI-DEE plays an important role in enhancing the stability of the Li||NMC811 cell with a capacity retention of ≈94% after 200 cycles under a high current density of 2.5 mA cm-2 . In addition, the 3.5 m LiFSI-DEE electrolyte exhibits good performance with anode-free batteries. This study offers a promising approach to enable ether-based electrolytes for high-voltage LMBs applications.

Design of a LiF-Rich Solid Electrolyte Interphase Layer through Highly Concentrated LiFSI-THF Electrolyte for Stable Lithium Metal Batteries.

Lithium metal is a promising anode material for lithium metal batteries (LMBs). However, dendrite growth and limited Coulombic efficiency (CE) during cycling have prevented its practical application in rechargeable batteries. Herein, a highly concentrated electrolyte composed of an ether solvent and lithium bis(fluorosulfonyl)imide (LiFSI) salt is introduced, which enables the cycling of a lithium metal anode at a high CE (up to ≈99%) without dendrite growth, even at high current densities. Using 3.85 m LiFSI in tetrahydrofuran (THF) as the electrolyte, a Li||Li symmetric cell can be cycled at 1.0 mA cm-2 for more than 1000 h with stable polarization of ≈0.1 V, and Li||LFP cells can be cycled at 2 C (1 C = 170 mA g-1 ) for more than 1000 cycles with a capacity retention of 94.5%. These excellent performances are observed to be attributed to the increased cation-anion associated complexes, such as contact ion pairs and aggregate in the highly concentrated electrolyte; revealed by Raman spectroscopy and theoretical calculations. These results demonstrate the benefits of a high-concentration LiFSI-THF electrolyte system, generating new possibilities for high-energy-density rechargeable LMBs.

Simultaneous Stabilization of the Solid/Cathode Electrolyte Interface in Lithium Metal Batteries by a New Weakly Solvating Electrolyte.

So far, the practical application of Li metal batteries has been hindered by the undesirable formation of Li dendrites and low Coulombic efficiencies (CEs). Herein, 1,2-diethoxyethane (DEE) is proposed as a new electrolytic solvent for lithium metal batteries (LMBs), and the performances of 1.0 m LiFSI in DEE are evaluated. Because of the low dielectric constant and dipole moment of DEE, the majority of the FSI- exists in associated states like contact ion pairs and aggregates, which is similar to the highly concentrated electrolytes. These associated complexes are involved in the reduction reaction on the Li metal anode, forming sound solid electrolyte interphase layers. Furthermore, free FSI- ions in DEE are observed to participate in the formation of cathode electrolyte interphase layers. These passivation layers not only suppress dendrite growth on the Li anode but also prevent unwanted side-reactions on the LiFePO4 cathode. The average CE of the Li||Cu cells in LiFSI-DEE is observed to be 98.0%. Moreover, LiFSI-DEE also plays an important role in enhancing the cycling stability of the Li||LiFP cell with a capacity retention of 93.5% after 200 cycles. These results demonstrate the benefits of LiFSI-DEE, which creates new possibilities for high-energy-density rechargeable LMBs.

Effect of ultrasound on removal of persistent organic pollutants (POPs) from different types of soils.

A new and promising technology is utilization of sonochemistry on decontamination of polluted soil. The feasibility of this technology on treatment of contaminated soils (synthetic clay, natural farm clay, and kaolin) was studied by using two target persistent organic pollutants (POPs): hexachlorobenzene (HCB) and phenanthrene (PHE). The soils were highly contaminated in 500 mg/kg. The laboratory experiments were conducted with various conditions (moisture, power, and time duration). The effects of these parameters on ultrasonication (as well as the removal of contaminants) were examined. The reasonable moisture ratio of the slurry could be in range of 2:1-3:1. The process did not change pH values of soils. Experimental results showed that ultrasonication has a potential to reduce the high concentrations of these POPs.