Revitalization of Diluent Amide-Based Electrolyte for Building High-Voltage Lithium-Metal Batteries.

Hitherto, highly concentrated electrolyte is the overarching strategy for revitalizing the usage of amide - in lithium-metal batteries (LMBs), which simultaneously mitigates the reactivity of amide toward Li and regulates uniform Li deposition via forming anion-solvated coordinate structure. However, it is undeniable that this would bring the cost burden for practical electrolyte preparation, which stimulates further electrolyte design toward tailoring anion-abundant Li+ solvation structure in stable amide electrolytes under a low salt content. Herein, a distinct method is conceived to design anions-enriched Li+ solvation structure in dilute amide-electrolyte (1 m Li-salt concentration) with the aid of integrating perfluoropolyethers (PFPE-MC) with anion-solvating ability and B/F-involved additives. The optimized electrolyte based on N,N-Dimethyltrifluoroacetamide (FDMAC) exhibits outstanding compatibility with Li and NCM622 cathode, facilitates uniform Li deposition along with robust solid electrolyte interphase (SEI) formation. Accordingly, both the lab-level LMB coin cell and practical pouch cell based on this dilute FDMAC electrolyte deliver remarkable performances with improved capacity and cyclability. This work pioneers the feasibility of diluted amide as electrolyte in LMB, and provides an innovative strategy for highly stable Li deposition via manipulating solvation structure within diluent electrolyte, impelling the electrolyte engineering development for practical high-energy LMBs.

Identification of a xylose-inducible promoter and its application for improving vitamin B12 production in Sinorhizobium meliloti.

Sinorhizobium meliloti 320 is a vitamin B12 (VB12 ) high-producing strain that has been isolated and identified in our previous study. Because the regulatory toolbox for S. meliloti is limited, we searched for new genetic components and identified the two xylose-inducible promoters PA and PB based on a promoter-probe vector with a green fluorescent protein (GFP) as reporter. Compared with the ParaA promoter from S. meliloti, both promoters exhibited higher induced expression and lower basal expression. Subsequently, the influence of glucose or sucrose on the expression of GFP driven by these three promoters was assayed. Glucose repressed all three promoters, and the expression of ParaA was the lowest in the presence of glucose. Although sucrose repressed the expression of PA by 35% and improved the expression of ParaA by 16%, the expression level of PA was the highest and was 13% higher than that of ParaA . Lastly, we overexpressed the hemA gene in the C4 pathway using the PA promoter in S. meliloti 320, and the VB12 production of the engineered strain increased by 11%. The VB12 production was further increased by 11% by adding 0.1% sodium succinate to the culture medium.

Application of different types of CRISPR/Cas-based systems in bacteria.

As important genome editing tools, CRISPR/Cas systems, especially those based on type II Cas9 and type V Cas12a, are widely used in genetic and metabolic engineering of bacteria. However, the intrinsic toxicity of Cas9 and Cas12a-mediated CRISPR/Cas tools can lead to cell death in some strains, which led to the development of endogenous type I and III CRISPR/Cas systems. However, these systems are hindered by complicated development and limited applications. Thus, further development and optimization of CRISPR/Cas systems is needed. Here, we briefly summarize the mechanisms of different types of CRISPR/Cas systems as genetic manipulation tools and compare their features to provide a reference for selecting different CRISPR/Cas tools. Then, we show the use of CRISPR/Cas technology for bacterial strain evolution and metabolic engineering, including genome editing, gene expression regulation and the base editor tool. Finally, we offer a view of future directions for bacterial CRISPR/Cas technology.

O2 Adsorption Associated with Sulfur Vacancies on MoS2 Microspheres.

MoS2 is well-known for its catalytic properties, mainly to adsorb hydrogenous or carbonaceous materials. However, the effect of MoS2 on the oxygen adsorption has been investigated only a few times thus far. In this work, we first studied the adsorbability of O2 by MoS2 through the analysis of Li2O2 growth on the surface of flower-like MoS2 microspheres with different concentrations of sulfur vacancies, which can be applied as the highly active electrocatalysts for Li-O2 batteries. The enhancement of battery performance for the Def-MoS2@CTs (CTs = carbon textile substrates) with a larger concentration of sulfur vacancies (S/Mo = 1.61) can be achieved. The experimental and theoretical results confirm that the sulfur vacancies play a crucial role in the adsorption process and thus affect the morphology and nucleation of Li2O2. In addition, a fundamental catalytic mechanism for this adsorption process is also proposed. These results provide a new insight into the development of a highly active electrocatalyst by introducing a large concentration of defects for Li-O2 batteries.

Pinpointing the L-phenylalanine binding sites of TyrR using biosensors and computer-aided simulation.

OBJECTIVES: To determine the binding sites for L-phenylalanine in TyrR protein via a rational mutation analysis combining biosensors and computer-aided simulation. RESULTS: TyrR protein of Escherichia coli is the chief transcriptional regulator of several genes essential for the biosynthesis and transport of aromatic amino acids. The identification of ligand-binding sites is often the starting point for protein function annotation and structure-based protein design. Here we combined computer-aided prediction methods and biosensors to identify the ligand-binding sites for L-Phe in TyrR protein. CONCLUSIONS: Residues at positions 160, 173 and 184 of TyrR protein are important for transcriptional activation of target genes tyrP induced by L-Phe, which indicates that they are the bona fide L-Phe binding sites of TyrR protein.

The distribution and function characterization of the i type lysozyme from Apostichopus japonicus.

Lysozyme is a very important component of the innate immune system and a key molecule that protects against bacterial infection. Sea cucumber i-type lysozyme (Aj-iLys) has been shown to possess multiple functions. In this study, we investigated the function and characterization of Aj-iLys in detail. Spatial distribution analysis showed that Aj-iLys was constitutively expressed in all tested tissues, with dominant expression in the tentacles and respiratory trees. Challenge with the pathogen V. splendidus and LPS stimulation both significantly up-regulated the mRNA expression of Aj-iLys. More importantly, inhibition of Aj-iLys expression by mRNA interference resulted in significant promotion of coelomocyte apoptosis during LPS challenge in vitro. The results indicated that Aj-iLys serves as an important innate immunity factor and plays a key defense role during host-pathogen interactions in sea cucumbers. From the radius of the antimicrobial zone, it was determined that the non-fusion Aj-iLys exerted a remarkable inhibitive effect on tested bacteria in vitro. Functional investigation revealed that Aj-iLys also exhibited isopeptidase activity based on its ability to hydrolyze l-Glutamic acid γ-(4-nitroanilide) in vitro to produce p-NA, which is an analogue of the isopeptide bond. The optimal catalytic conditions for the isopeptidase activity were 37 °C, pH 6.5, and the optimum ionic strength was about 0.050 mol/L.

Rational design and analysis of an Escherichia coli strain for high-efficiency tryptophan production.

L-tryptophan (L-trp) is a precursor of various bioactive components and has great pharmaceutical interest. However, due to the requirement of several precursors and complex regulation of the pathways involved, the development of an efficient L-trp production strain is challenging. In this study, Escherichia coli (E. coli) strain KW001 was designed to overexpress the L-trp operator sequences (trpEDCBA) and 3-deoxy-D-arabinoheptulosonate-7-phosphate synthase (aroG fbr ). To further improve the production of L-trp, pyruvate kinase (pykF) and the phosphotransferase system HPr (ptsH) were deleted after inactivation of repression (trpR) and attenuation (attenuator) to produce strain KW006. To overcome the relatively slow growth and to increase the transport rate of glucose, strain KW018 was generated by combinatorial regulation of glucokinase (galP) and galactose permease (glk) expression. To reduce the production of acetic acid, strain KW023 was created by repressive regulation of phosphate acetyltransferase (pta) expression. In conclusion, strain KW023 efficiently produced 39.7 g/L of L-trp with a conversion rate of 16.7% and a productivity of 1.6 g/L/h in a 5 L fed-batch fermentation system.

[Emodin-induced increase in expression of ß1 subunit of BKCa channel mediates relaxation of cerebral basilar artery in spontaneously hypertensive rats].

The purposes of this study were to investigate the effect of emodin on expression of BKCa channel ß1 subunit in basilar artery smooth muscle cells (BASMCs) and electrophysiological characteristics of vascular smooth muscle cells in spontaneously hypertensive rats (SHR). Tail artery pressure measurement instrument was used to measure the change of SHR systolic blood pressure before and after emodin intervention. Single vascular smooth muscle cell was electrically recorded by whole-cell patch-clamp technique. Immunohistochemistry and Western blotting were used to study the distribution and expression of the BKCa channel ß1 subunit. The results showed that emodin decreased blood pressure of SHR from (223 ± 16) mmHg to (127 ± 12) mmHg (P < 0.01). There was no difference of blood pressure between emodin-treated SHR and Wistar rats. Emodin significantly increased outward currents of smooth muscle cells in SHR (P < 0.05), and this effect could be reversed by specific inhibitor of BKCa channel, IbTX. Emodin also up-regulated BKCa channel ß1 subunit expression in BASMCs. These results suggest that emodin relaxes cerebral basilar artery by enhancing BKCa current via increasing ß1 subunit expression in BASMCs.

Structurally integrated asymmetric polymer electrolyte with stable Janus interface properties for high-voltage lithium metal batteries.

Solid-state polymer electrolytes are outstanding candidates for next-generation lithium metal batteries in the realm of high specific energy densities, high safeties and tight contact with electrodes. However, their applications are still hindered by the limitations that no single polymer is electrochemically stable with the oxidizing high-voltage cathode and the highly reductive Li anode, simultaneously. Herein, a bilayer asymmetric polymer electrolyte (SL-SPE) without accessional interface resistance that using poly (ethylene glycol) diacrylate (PEGDA) as a "bridge" to connect the sulfonyl (OS = O)-contained oxidation-tolerated layer and polyether-derived reduction-tolerated layer (SPE), is proposed and synthesized by sequential two-step UV polymerizations. SL-SPE can provide widened electrochemical stability window up to 5 V, while simultaneously deploying a stable Janus interface property. Eventually, the superior high-voltage (4.4 V) cycling durability can be displayed in LiNi0.6Co0.2Mn0.2O2|SL-SPE|Li batteries. This finding provides a bran-new idea for designing multifunctional polymer electrolytes in the application of solid-state batteries.

Tackling the Interfacial Issues of Spinel LiNi0.5Mn1.5O4 by Room-Temperature Spontaneous Dediazonation Reaction.

The detrimental interfacial side reactions, inducing electrolyte decomposition and transition-metal dissolution, are regarded as "arch-criminal" for the utilization of spinel LiNi0.5Mn1.5O4 (LNMO) in high-power Li-ion batteries (LIBs). To conquer this issue, herein, we construct a thin polyphenyl film onto the surface of LNMO via the spontaneous dediazonation of C6H5N2+BF4- at room temperature. This conductive film facilitates the Li+ transport within cathode and at LNMO|electrolyte interface while reinforcing the compatibility of LNMO against electrolyte by efficiently suppressing the electrolyte decomposition catalyzed by LNMO and even the transition-metal dissolution. Consequently, polyphenyl-grafted LNMO exhibits improved electrochemical performances, e.g., the considerable discharge capacity of 136.7 mAh g-1 at low current density (0.1C), excellent rate capability, and long-term cyclability with a reversible capacity of 107.4 mAh g-1 along with high capacity retention of ∼85% after cycling 500 times, that are superior to those of the pristine LNMO counterpart. All these results demonstrate that our strategy is instrumental in solving the interface issues with respect to the spinel LNMO cathode, impelling the development of LNMO-based batteries with high energy density.

Recent Insights Into the Prognostic and Therapeutic Applications of Lysozymes.

Lysozymes are naturally occurring enzymes present in a variety of biological organisms, such as bacteria, fungi, and animal bodily secretions and tissues. It is also the main ingredient of many ethnomedicines. It is well known that lysozymes and lysozyme-like enzymes can be used as anti-bacterial agents by degrading bacterial cell wall peptidoglycan that leads to cell death, and can also inhibit fungi, yeasts, and viruses. In addition to its direct antimicrobial activity, lysozyme is also an important component of the innate immune system in most mammals. Increasing evidence has shown the immune-modulatory effects of lysozymes against infection and inflammation. More recently, studies have revealed the anti-cancer activities of lysozyme in multiple types of tumors, potentially through its immune-modulatory activities. In this review, we summarized the major functions and underlying mechanisms of lysozymes derived from animal and plant sources. We highlighted the therapeutic applications and recent advances of lysozymes in cancers, hypertension, and viral diseases, aiming toseeking alternative therapies for standard medical treatment bypassing side effects. We also evaluated the role of lysozyme as a promising cancer marker for prognosis to indicate the outcomes recurrence for patients.

Nitrogen-sulfur dual-doped porous carbon spheres/sulfur composites for high-performance lithium-sulfur batteries.

A nitrogen-sulfur dual-doped porous carbon spheres/sulfur composite (PCS-NS/S) sample was prepared by a simple hydrothermal method with starch and l-methionine as carbon and nitrogen-sulfur resources, respectively. XRD, XPS, and N2 adsorption-desorption tests were used to characterize the crystal and pore structure of the PCS-NS/S sample. The morphology and weight ratio of sulfur were investigated by SEM, TEM, and TG analyses. The sample was used as the positive electrode for lithium-sulfur batteries and found to exhibit excellent electrochemical performance.

Binary Mixtures of Highly Concentrated Tetraglyme and Hydrofluoroether as a Stable and Nonflammable Electrolyte for Li-O2 Batteries.

Developing a long-term stable electrolyte is one of the most enormous challenges for Li-O2 batteries. Equally, the high flammability of frequently used solvents seriously weakens the electrolyte safety in Li-O2 batteries, which inevitably restricts their commercial applications. Here, a binary mixture of highly concentrated tetraglyme electrolyte (HCG4) and 1,1,2,2-tetrafluoroethyl 2,2,3,3-tetrafluoropropyl ether (TTE) was used for a novel electrolyte (HCG4/TTE) in Li-O2 batteries, which exhibit good wettability, enhanced ionic conductivity, considerable nonflammability, and high electrochemical stability. Being a co-solvent, TTE can contribute to increasing ionic conductivity and to improving flame retardance of the as-prepared electrolyte. The cell with this novel electrolyte displays an enhanced cycling stability, resulting from the high electrochemical stability during cycling and the formation of electrochemically stable interfaces prevents parasitic reactions occurring on the Li anode. These results presented here demonstrate a novel electrolyte with a high electrochemical stability and considerable safety for Li-O2 batteries.

Self-assembled layer-by-layer partially reduced graphene oxide-sulfur composites as lithium-sulfur battery cathodes.

Constructing a reliable conductive carbon matrix is essential for the sulfur-containing cathode materials of lithium-sulfur batteries. A ready-made conductive matrix infiltrated with sulfur as the cathode is the usual solution. Here, a partially reduced graphene oxide-sulfur composite (prGO/S) with an ordered self-assembled layer-by-layer structure is introduced as a Li-S battery cathode. The prGO/S composites are synthesized through a facile one-step self-assembly liquid route. An appropriate amount of sulfur is in situ deposited on the surface of the prGO nanosheets by adjusting the reduction degree of the GO nanosheets. The combined effect of the electrostatic repulsions and surface energy makes the sulfur wrapped prGO nanosheets self-assemble to form an ordered layer-by-layer structure, which not only ensures the uniform distribution of sulfur but also accommodates the volume change of the sulfur species during cycling. Moreover, the conductivity of the prGO/S composites improves when the reduction time increases. XPS spectra confirm that sulfur is still chemically bonded to the prGO. After applying the prGO coating of the prGO/S composite particle and as an interlayer in a lithium-sulfur battery configuration, a high initial discharge capacity of 1275.8 mA h g-1 is achieved and the discharge capacity of the 100th cycle is 1013.8 mA h g-1 at 0.1C rate.

[Recombinant expression and antibacterial activity of i-type lysozyme from sea cucumber Stichopus japonicus].

The cDNA of an i type lysozyme was cloned from Stichopus japonicus (named as SjLys). The DNA fragment of the mature SjLys was subcloned into expression vector of pET-32a (+) to construct the recombinant plasmid of pET32a (+)-SjLys. The recombinant plasmid was then transformed into Escherichia coli BL21 (DE3) pLysS and induced by isopropylthio-beta-D-galactoside (IPTG). The recombinant protein expressed as inclusion bodies was denatured, partially purified and refolded to be an active form. The bacteriolytic activity of recombinant protein purified by the metal-chelating was 19.2 U/mg. The antibacterial activity of the purified recombinant SjLys (rSjLys) was analyzed. The rSjLys protein displayed inhibitive effect on the growth of the tested Gram-positive and Gram-negative bacteria. In particular, rSjLys had a strong inhibitive activity on Vibrio parahaemolyticus and Pseudomonas aeruginosa, both the most common pathogenic bacteria in the marine animals. The heat-treated rSjLys exhibited more potent activities against all tested bacteria. These results indicated that the S. japonicus lysozyme was the enzyme with combined enzymatic (glycosidase) and non-enzymatic antibacterial action, and it had a wide antibacterial spectrum. Therefore, it is suggested that the S. japonicus lysozyme should be one of the important molecules against pathogens in the innate immunity of sea cucumbers.

Characterization of an i-type lysozyme gene from the sea cucumber Stichopus japonicus, and enzymatic and nonenzymatic antimicrobial activities of its recombinant protein.

Because sea cucumbers lack a well-developed immune system and can ingest pathogenic bacteria together with food, some form of active antibacterial substances must be present in the body for defense. In this study, the cDNA of an i-type lysozyme from the sea cucumber Stichopus japonicus (designated SjLys) was cloned by RT-PCR and RACE PCR techniques. The full length cDNA of SjLys was 713 bp with an open reading frame of 438 bp coding for 145 amino acids. Two catalytic residues (Glu34 and Asp47), conserved in i-type lysozymes, and a highly conserved region near the active site, MDVGSLSCG(P\Y)(Y\F)QIK, were detected in SjLys. In addition, the domain structure analysis of SjLys showed that it is highly similar to the medicinal leech destabilase, which belongs to a new phylogenetic family of invertebrate lysozymes possessing both glycosidase and isopeptidase activities. To gain insight into the in vitro antimicrobial activities of SjLys, the mature peptide coding region was heterologously expressed in Escherichia coli. The recombinant SjLys protein displayed an inhibitive effect on the growth of the tested Gram-positive and Gram-negative bacteria. A remarkable finding is that the recombinant SjLys exhibited more potent activities against all tested bacterial strains after heat-treating at 100 degrees C for 50 min. These results indicated that the S. japonicus lysozyme is an enzyme with combined enzymatic (glycosidase) and nonenzymatic antibacterial action.

High-level soluble expression of the functional peptide derived from the C-terminal domain of the sea cucumber lysozyme and analysis of its antimicrobial activity

Background The sea cucumber lysozyme belongs to the family of invertebrate lysozymes and is thought to be a key defense factor in protecting aquaculture animals against bacterial infection. Recently, evidence was found that the sea cucumber lysozyme exerts broad spectrum antimicrobial action in vitro against Gram-negative and Gram-positive bacteria, and it also has more potent antimicrobial activity independent of its enzymatic activity. To explore the antimicrobial role of this non-enzymatic lysozyme and model its structure to novel antimicrobial peptides, the peptide from the C-terminal amino acid residues 70-146 of the sea cucumber lysozyme in Stichopus japonicus (SjLys-C) was heterologously expressed in Escherichia coli Rosetta(DE3)pLysS. Results The fusion protein system led to over-expression of the soluble and highly stable product, an approximate 26 kDa recombinant SjLys-C protein (rSjLys-C). The present study showed that rSjLys-C displayed strong antimicrobial activity against the tested Gram-positive and Gram-negative bacteria. In particular, the heat-treated rSjLys-C exhibited more inhibitive activity than the native rSjLys-C. The structural analysis of SjLys-C showed that it is a typical hydrophilic peptide and contains a helix-loop-helix motif. The modeling of SjLys-C molecular structures at different temperatures revealed that the tertiary structure of SjLys-C at 100°C underwent a conformational change which is favorable for enhancing antimicrobial activity. Conclusion These results indicate that the expressed rSjLys-C is a highly soluble product and has a strong antimicrobial activity. Therefore, gaining a large quantity of biologically active rSjLys-C will be used for further biochemical and structural studies and provide a potential use in aquaculture and medicine.

Cloning and characterization of genes encoded in dTDP-D-mycaminose biosynthetic pathway from a midecamycin-producing strain, Streptomyces mycarofaciens.

Two subclusters from Streptomyces mycarofaciens, a midecamycin producer, were cloned and partially sequenced. One region was located at the 5' end of the mid polyketide synthase (PKS) genes and contained the genes midA, midB and midC. The other region was at the 3' end of the PKS genes and contained midK, midI and midH. Analysis of the nucleotide sequence revealed that these genes encode dTDP-glucose synthase (midA), dTDP-glucose dehydratase (midB), aminotransferase (midC), methyltransferase (midK), glycosyltransferase (midI) and an assistant gene (midH). All of these genes are involved in the biosynthesis of dTDP-D-mycaminose, the first deoxysugar of midecamycin, and in transferring the mycaminose to the midecamycin aglycone in S. mycarofaciens. Similar to gene pairs desVIII/desVII in S. venezuelae and tylMIII/tylMII in S. fradiae, the product of midH probably functions as an auxiliary protein required by the MidI protein for efficient glycosyltransfer in midecamycin biosynthesis.