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This study addresses the environmental and resource challenges posed by the growing volume of waste electric porcelain in the power industry by developing solar absorption and thermal storage integrated ceramics (SATS ceramics) from waste electric porcelain. These SATS ceramics, which feature an exceptional mullite structure, were crafted through the optimization of the sintering process. Notably, when sintered at 1400 °C with an 11 wt.% magnetite content, the resulting material boasts an optimal short-clustered, ordered, and interwoven columnar mullite structure. This structure endows the material with a remarkable flexural strength of 96.05 MPa and a specific heat capacity of up to 0.6415 J/(g* °C) at 300 °C, significantly enhancing its thermal energy storage efficiency. This research offers innovative insights into the high-value utilization of waste electric porcelain and the development of solar thermal storage materials, underscoring its significant environmental and economic advantages.

Abstract Background Natural life systems can be significantly modified at the genomic scale by human intervention, demonstrating the great innovation capacity of genome engineering. Large epi-chromosomal DNA structures were established in Escherichia coli cells, but some of these methods were inconvenient, using heterologous systems, or relied on engineered E. coli strains. Results The wild-type model bacterium E. coli has a single circular chromosome. In this work, a novel method was developed to split the original chromosome of wild-type E. coli. With this method, novel E. coli strains containing two chromosomes of 0.10 Mb and 4.54 Mb, and 2.28 Mb and 2.36 Mb were created respectively, designated as E. coli 0.10/4.54 and E. coli 2.28/2.36. The new chromosomal arrangement was proved by PCR amplification of joint regions as well as a combination of Nanopore and Illumina sequencing analysis. While E. coli 0.10/4.54 was quite stable, the two chromosomes of E. coli 2.28/2.36 population recombined into a new chromosome (Chr.4.64MMut), via recombination. Both engineered strains grew slightly slower than the wild-type, and their cell shapes were obviously elongated. Conclusion Finally, we successfully developed a simple CRISPR-based genome engineering technique for the construction of multi-chromosomal E. coli strains with no heterologous genetic parts. This technique might be applied to other prokaryotes for synthetic biology studies and applications in the future.

Abstract Glucose and xylose are two major components of lignocellulose. Simultaneous consumption of glucose and xylose is critical for engineered microorganisms to produce fuels and chemicals from lignocellulosic biomass. Although many production limitations have been resolved, glucose‐induced inhibition of xylose transport remains a challenge. In this study, a cell growth‐based screening strategy was designed to identify xylose transporters uninhibited by glucose. The glucose pathway was genetically blocked in Escherichia coli so that glucose functions only as an inhibitor and cells need xylose as the carbon source for survival. Through adaptive evolution, omics analysis and reverse metabolic engineering, a new phosphoenolpyruvate: carbohydrate phosphotransferase system (PTS) galactitol transporter (GalABC, encoded by EcolC_1640, EcolC_1641, and EcolC_1642 genes) that is not inhibited by glucose was identified. Inactivation of adenylate cyclase led to increased expression of the EcolC_1642 gene, and a point mutation in gene EcolC_1642 (N13S) further enhanced xylose transport. During the second round of gene mining, AraE and a new ABC transporter (AraFGH) of xylose were identified. A point mutation in the transcription regulator araC (L156I) caused increased expression of araE and araFGH genes without arabinose induction, and a point mutation in araE (D223Y) further enhanced xylose transport. These newly identified xylose transporters can support the simultaneous consumption of glucose and xylose and have potential use in producing chemicals from lignocellulose.

Abstract Synthetic biology has been represented by the creation of artificial life forms at the genomic scale. In this work, a CRISPR‐based chromosome‐doubling technique is designed to first construct an artificial diploid Escherichia coli cell. The stable single‐cell diploid E. coli is isolated by both maximal dilution plating and flow cytometry, and confirmed with quantitative PCR, fluorescent in situ hybridization, and third‐generation genome sequencing. The diploid E. coli has a greatly reduced growth rate and elongated cells at 4–5 µm. It is robust against radiation, and the survival rate after exposure to UV increased 40‐fold relative to WT. As a novel life form, the artificial diploid E. coli is an ideal substrate for research fundamental questions in life science concerning polyploidy. And this technique may be applied to other bacteria.

Abstract Background Tailoring gene expression to balance metabolic fluxes is critical for the overproduction of metabolites in yeast hosts, and its implementation requires coordinated regulation at both transcriptional and translational levels. Although synthetic minimal yeast promoters have shown many advantages compared to natural promoters, their transcriptional strength is still limited, which restricts their applications in pathway engineering. Results In this work, we sought to expand the application scope of synthetic minimal yeast promoters by enhancing the corresponding translation levels using specific Kozak sequence variants. Firstly, we chose the reported UASF-E-C-Core1 minimal promoter as a library template and determined its Kozak motif (K0). Next, we randomly mutated the K0 to generate a chimeric promoter library, which was able to drive green fluorescent protein (GFP) expression with translational strengths spanning a 500-fold range. A total of 14 chimeric promoters showed at least two-fold differences in GFP expression strength compared to the K0 control. The best one named K528 even showed 8.5- and 3.3-fold increases in fluorescence intensity compared with UASF-E-C-Core1 and the strong native constitutive promoter P TDH3 , respectively. Subsequently, we chose three representative strong chimeric promoters (K540, K536, and K528) from this library to regulate pathway gene expression. In conjunction with the tHMG1 gene for squalene production, the K528 variant produced the best squalene titer of 32.1 mg/L in shake flasks, which represents a more than 10-fold increase compared to the parental K0 control (3.1 mg/L). Conclusions All these results demonstrate that this chimeric promoter library developed in this study is an effective tool for pathway engineering in yeast.

Summary The unconventional yeast Pichia kudriavzevii is renowned for its ability to survive at low pH and has been exploited for the industrial production of various organic acids, especially succinic acid (SA). However, P. kudriavzevii can also utilize the di‐ and tricarboxylate intermediates of the Krebs cycle as the sole carbon sources for cell growth, which may adversely affect the extracellular accumulation of SA. Because the carboxylic acid transport machinery of P. kudriavzevii remains poorly understood, here, we focused on studying its SA transportation process from the perspective of mining and characterization of dicarboxylate transporters in a newly isolated acid‐tolerant P. kudriavzevii strain CY902. Through genome sequencing and transcriptome analysis, two JEN family carboxylate transporters (PkJEN2‐1 and PkJEN2‐2) were found to be involved in SA transport. Substrate specificity analysis revealed that both PkJEN proteins are active dicarboxylate transporters, that can effectively import succinate, fumarate and L‐malate into the cell. In addition, PkJEN2‐1 can transport α‐ketoglutarate, while PkJEN2‐2 cannot. Since PkJEN2‐1 shows higher transcript abundance than PkJEN2‐2, its role in dicarboxylate transport is more important than PkJEN2‐2. In addition, PKJEN2‐2 is also responsible for the uptake of citrate. To our best knowledge, this is the first study to show that a JEN2 subfamily transporter is involved in tricarboxylate transport in yeast. A combination of model‐based structure analysis and rational mutagenesis further proved that amino acid residues 392‐403 of the tenth transmembrane span (TMS‐X) of PkJEN2‐2 play an important role in determining the specificity of the tricarboxylate substrate. Moreover, these two PkJEN transporters only exhibited inward transport activity for SA, and simultaneous inactivation of both PkJEN transporters reduced the SA influx, resulting in enhanced extracellular accumulation of SA in the late stage of fermentation. This work provides useful information on the mechanism of di‐/tricarboxylic acid utilization in P. kudriavzevii, which will help improve the organic acid production performance of this microbial chassis.

Aspergillus, as a genus of filamentous fungi, has members that display a variety of different behavioural strategies, which are affected by various environmental factors. The decoded genomic sequences of many species vary greatly in their evolutionary similarities, encouraging studies on the functions and evolution of the Aspergillus genome in complex natural environments. Here, we present the 26 Mb de novo assembled high-quality reference genome of Aspergillus glaucus ‘China Changchun halophilic Aspergillus’ (CCHA), which was isolated from the surface of plants growing near a salt mine in Jilin, China, based on data from whole-genome shotgun sequencing using Illumina Solexa technology. The sequence, coupled with data from comprehensive transcriptomic survey analyses, indicated that the redox state and transmembrane transport might be critical molecular mechanisms for the adaptation of A. glaucus ‘CCHA’ to the high-salt environment of the saltern. The isolation of salt tolerance-related genes, such as CCHA-2114, and their overexpression in Escherichia coli demonstrated that A. glucus ‘CCHA’ is an excellent organism for the isolation and identification of salt tolerant-related genes. These data expand our understanding of the evolution and functions of fungal and microbial genomes, and offer multiple target genes for crop salt-tolerance improvement through genetic engineering.

D-glycerate is an attractive chemical for a wide variety of pharmaceutical, cosmetic, biodegradable polymers, and other applications. Now several studies have been reported about the synthesis of glycerate by different biotechnological and chemical routes from glycerol or other feedstock. Here, we present the construction of an Escherichia coli engineered strain to produce optically pure D-glycerate by oxidizing glycerol with an evolved variant of alditol oxidase (AldO) from Streptomyces coelicolor. This is achieved by starting from a previously reported variant mAldO and employing three rounds of directed evolution, as well as the combination of growth-coupled high throughput selection with colorimetric screening. The variant eAldO3-24 displays a higher substrate affinity toward glycerol with 5.23-fold than the wild-type AldO, and a 1.85-fold increase of catalytic efficiency (kcat/KM). Then we introduced an isopropyl-β-D-thiogalactopyranoside (IPTG)-inducible T7 expression system in E. coli to overexpress the variant eAldO3-24, and deleted glucosylglycerate phosphorylase encoding gene ycjM to block the consumption of D-glycerate. Finally, the resulting strain TZ-170 produced 30.1 g/l D-glycerate at 70 h with a yield of 0.376 mol/mol in 5-l fed-batch fermentation. [ABSTRACT FROM AUTHOR]

Background: Escherichia coli suffer from osmotic stress during succinic acid (SA) production, which reduces the performance of this microbial factory. Results: Here, we report that a point mutation leading to a single amino acid change (D654Y) within the β-subunit of DNA-dependent RNA polymerase (RpoB) significantly improved the osmotolerance of E. coli. Importation of the D654Y mutation of RpoB into the parental strain, Suc-T110, increased cell growth and SA production by more than 40% compared to that of the control under high glucose osmolality. The transcriptome profile, determined by RNA-sequencing, showed two distinct stress responses elicited by the mutated RpoB that counterbalanced the osmotic stress. Under non-stressed conditions, genes involved in the synthesis and transport of compatible solutes such as glycine-betaine, glutamate or proline were upregulated even without osmotic stimulation, suggesting a "pre-defense" mechanism maybe formed in the rpoB mutant. Under osmotic stressed conditions, genes encoding diverse sugar transporters, which should be down-regulated in the presence of high osmotic pressure, were derepressed in the rpoB mutant. Additional genetic experiments showed that enhancing the expression of the mal regulon, especially for genes that encode the glycoporin LamB and maltose transporter, contributed to the osmotolerance phenotype. Conclusions: The D654Y single amino acid substitution in RpoB rendered E. coli cells resistant to osmotic stress, probably due to improved cell growth and viability via enhanced sugar uptake under stressed conditions, and activated a potential "pre-defense" mechanism under non-stressed conditions. The findings of this work will be useful for bacterial host improvement to enhance its resistance to osmotic stress and facilitate bio-based organic acids production. [ABSTRACT FROM AUTHOR]

Background: Lignocellulase hypersecretion has been achieved in industrial fungal workhorses such as Trichoderma reesei, but the underlying mechanism associated with this process is not well understood. Although previous comparative genomic studies have revealed that the mutagenic T. reesei strain RUT-C30 harbors hundreds of mutations compared with its parental strain QM6a, how these mutations actually contribute to the hypersecretion phenotype remains to be elucidated. Results: In this study, we systematically screened gene knockout (KO) mutants in the cellulolytic fungus Neurospora crassa, which contains orthologs of potentially defective T. reesei RUT-C30 mutated genes. Of the 86 deletion mutants screened in N. crassa, 12 exhibited lignocellulase production more than 25% higher than in the wild-type (WT) strain and 4 showed nearly 25% lower secretion. We observed that the deletion of Ncap3m (NCU03998), which encodes the µ subunit of the adaptor protein 3 (AP-3) complex in N. crassa, led to the most significant increase in lignocellulase secretion under both Avicel and xylan culture conditions. Moreover, strains lacking the ß subunit of the AP-3 complex, encoded by Ncap3b (NCU06569), had a similar phenotype to ?Ncap3m, suggesting that the AP-3 complex is involved in lignocellulase secretion in N. crassa. We also found that the transcriptional abundance of major lignocellulase genes in ?Ncap3m was maintained at a relatively higher level during the late stage of fermentation compared with the WT, which might add to the hypersecretion phenotype. Finally, we found that importation of the T. reesei ap3m ortholog Trap3m into ?Ncap3m can genetically restore secretion of lignocellulases to normal levels, which suggests that the effect of the AP-3 complex on lignocellulase secretion is conserved in cellulolytic ascomycetes. Conclusions: Using the model cellulolytic fungus N. crassa, we explored potential hypersecretion-related mutations in T. reesei strain RUT-C30. Through systematic genetic screening of 86 corresponding orthologous KO mutants in N. crassa, we identified several genes, particularly those encoding the AP-3 complex that contribute to lignocellulase secretion. These findings will be useful for strain improvement in future lignocellulase and biomass-based chemical production. [ABSTRACT FROM AUTHOR]

Both mammalian tensin-like phosphatase 1 [TEP1; also known as phosphatase deleted on chromosome 10 (PTEN) or mutated in multiple advanced cancer 1 (MMAC1)] and Saccharomyces cerevisiae ScTep1p are involved in the phosphatidylinositol pathway. In this study, we identified the Fusarium graminearum locus FGSG_04982.3 (named FgTEP1) as the functional homologue of ScTEP1 in the sensitivity of S. cerevisiae cells to wortmannin, the phosphatidylinositol-3 kinase inhibitor. Deletion of FgTEP1 causes F. graminearum mycelial growth to become sensitive to lithium and reduces the production of conidia. Although conidia lacking FgTEP1 germinate normally, they show reduced germination efficiency in the presence of wortmannin. In addition, we showed that deletion of FgTEP1 reduces the virulence of F. graminearum on wheat. These results indicate that FgTep1p is linked to the phosphatidylinositol-3 kinase signalling pathway in this plant fungal pathogen. [ABSTRACT FROM AUTHOR]

Shrinkage is observed in many electrospun membranes. The stretched conformation of the macromolecular chains has been proposed as the possible cause. However, so far, our understanding of the fundamentals is still qualitative and cannot provide much help in the shrinkage control. In this paper, based on the crimped fibers after stimulus-induced shrinkage, a clear evidence of buckling, the gradient pre-strain field in the cross-section of the electrospun fibers, which is the result of a gradient solidification field and a tensile force in the fibers during electrospinning, is identified as the underlying mechanism for the stimulus-induced shrinkage. Subsequently, two buckling conditions are derived. Subsequently, a series of experiments are carried out to reveal the influence of four typical processing parameters (namely, the applied voltage, solution concentration, distance between electrodes, and rotation speed of collector), which are highly relevant to the formation of the gradient pre-strain field. It is concluded that there are some different ways to achieve the required shrinkage ratios in two in-plane directions (i.e., the rotational and transverse directions of the roller collector). Some of the combinations of these parameters are more effective at achieving high uniformity than others. Hence, it is possible to optimize the processing parameters to produce high-quality membranes with well-controlled shrinkage in both in-plane directions.

In this work, a novel flexible electrically resistive-type MXene/Thermoplastic polyurethanes(TPU) based strain sensors was developed by a composite process of electrospinning (ES) and electrostatic spray deposition (ESD). Compared with other deposition processes, the sensing layer prepared by ESD has better adhesion to the ES TPU nanofiber membrane and is not easy to crack during the stretching process, thereby greatly improving the working range of the strain sensor. Furthermore, we obtained the sandwich structure easily by ES on the surface of the sensing layer again. This will help make the stress distribution more uniform during the stretching process and further increase the strain sensing range. The ESD-ES strain sensors were attached on skin to monitor various human motions. The results demonstrate that our ESD-ES strain sensors have wide application prospects in smart wearable device.

Laser-induced graphene (LIG) has the advantages of one-step fabrication, prominent mechanical performance, as well as high conductivity; it acts as the ideal material to fabricate flexible strain sensors. In this study, a wearable flexible strain sensor consisting of three-dimensional (3D) wavy LIG and silicone rubber was reported. With a laser to scan on a polyimide film, 3D wavy LIG could be synthesized on the wavy surface of a mold. The wavy-LIG strain sensor was developed by transferring LIG to silicone rubber substrate and then packaging. For stress concentration, the ultimate strain primarily took place in the troughs of wavy LIG, resulting in higher sensitivity and less damage to LIG during stretching. As a result, the wavy-LIG strain sensor achieved high sensitivity (gauge factor was 37.8 in a range from 0% to 31.8%, better than the planar-LIG sensor), low hysteresis (1.39%) and wide working range (from 0% to 47.7%). The wavy-LIG strain sensor had a stable and rapid dynamic response; its reversibility and repeatability were demonstrated. After 5000 cycles, the signal peak varied by only 2.32%, demonstrating the long-term durability. Besides, its applications in detecting facial skin expansion, muscle movement, and joint movement, were discussed. It is considered a simple, efficient, and low-cost method to fabricate a flexible strain sensor with high sensitivity and structural robustness. Furthermore, the wavy-LIG strain senor can be developed into wearable sensing devices for virtual/augmented reality or electronic skin.

The electrospinning electric field is the main factor responsible for the jet stretching force. Previous studies showed that nonhomogeneous electric field would affect the jets formed during the multi-needles electrospinning. Therefore, a multitude of methods were explored to optimize the electric field in multi-needles electrospinning, such as adjusting the supplied voltage on each needle, varying the space of the needles and spatial distribution of the needles. In this paper, three categories of needles distribution were explored for a homogeneous electric field and the design of a higher needles density of electrospinning head. The theoretical and experimental results showed that the homogeneous electric field formed with the trapezoidal head at 15mm distance while the others’ head required over 30mm. Hence, the trapezoidal arrangement is a better multi-needles distribution for electrospinning head, and has much more significance for high yield of electrospinning nanofibers.

In this paper, we synergistically combine electrohydrodynamic (EHD) printing and replica molding for the fabrication of microlenses. Glycerol solution microdroplets was sprayed onto the ITO glass to form liquid mold by an EHD printing process. The liquid mold is used as a master to fabricate a polydimethylsiloxane (PDMS) mold. Finally, the desired micro-optical device can be fabricated on any substrate using a PDMS soft lithography mold. We demonstrate our strategy by generating microlenses of photocurable polymers and by characterizing their optical properties. It is a new method to rapidly and cost-effectively fabricate molds with small diameters by exploiting the advantages of EHD printing, while maintaining the parallel nature of soft-lithography.

Fusarium head blight (FHB) is a worldwide devastating disease of wheat, caused primarily by species in the Fusarium graminearum (Fg) complex. In this study, we obtained 55 Fusarium isolates from wheat with FHB collected from seven provinces along the north of the Yangtze River. One additional phylogenetic species of Fg complex, Fusarium meridionale, was identified for the first time from China in addition to two known ones, Fusarium asiaticum and F.  graminearum. In addition, Fusarium acuminatum, distantly related to Fg complex, was for the first time identified in Northern China. Sensitivities of these isolates to carbendazim were examined and appeared to vary both within and between species. Mycotoxin genotype analyses indicated that F.  asiaticum isolates were potential 3-AcDON and NIV mycotoxin producers, while all F.  graminearum isolates might be 15-AcDON producers. These findings would provide useful information for developing management strategies for the control of FHB in Northern China. [ABSTRACT FROM AUTHOR]

The fabrication of three-dimensional scaffolds has attracted more attention in tissue engineering. The purpose of this study is to explore a new method for the fabrication of three-dimensional micro-nanofiber structures by combining solution blow spinning and rotating collector. In this study, we successfully fabricated fibers with a minimum diameter of 200 nm and a three-dimensional structure with a maximum porosity of 89.9%. At the same time, the influence of various parameters such as the solvent volatility, the shape of the collector, the feed rate of the solution and the applied gas pressure were studied. It is found that solvent volatility has large effect on the formation of the three-dimensional shape of the structure. The shape of the collector affects the porosity and fiber distribution of the three-dimensional structure. The fiber diameter and fiber uniformity can be controlled by adjusting the solution feed rate and the applied gas pressure. It is feasible to fabricate high-quality three-dimensional micro-nanofiber structure by this new method, which has great potential in tissue engineering.

Improvement in the osmotolerance of Escherichia coli is essential for the production of high titers of various bioproducts. In this work, a cusS mutation that was identified in the previously constructed high-succinate-producing E. coli strain HX024 was investigated for its effect on osmotolerance. CusS is part of the twocomponent system CusSR that protects cells from Ag(I) and Cu(I) toxicity. Changing cusS from strain HX024 back to its original sequence led to a 24% decrease in cell mass and succinate titer under osmotic stress (12% glucose). When cultivated with a high initial glucose concentration (12%), introduction of the cusS mutation into parental strain Suc-T110 led to a 21% increase in cell mass and a 40% increase in succinate titer. When the medium was supplemented with 30 g/liter disodium succinate, the cusS mutation led to a 120% increase in cell mass and a 492% increase in succinate titer. Introducing the cusS mutation into the wild-type strain ATCC 8739 led to increases in cell mass of 87% with 20% glucose and 36% using 30 g/liter disodium succinate. The cusS mutation increased the expression of cusCFBA, and gene expression levels were found to be positively related to osmotolerance abilities. Because high osmotic stress has been associated with deleterious accumulation of Cu(I) in the periplasm, activation of CusCFBA may alleviate this effect by transporting Cu(I) out of the cells. This hypothesis was confirmed by supplementing sulfurcontaining amino acids that can chelate Cu(I). Adding methionine or cysteine to the medium increased the osmotolerance of E. coli under anaerobic conditions. IMPORTANCE In this work, an activating Cus copper efflux system was found to increase the osmotolerance of E. coli. In addition, new osmoprotectants were identified. Supplementation with methionine or cysteine led to an increase in osmotolerance of E. coli under anaerobic conditions. These new strategies for improving osmotolerance will be useful for improving the production of chemicals in industrial bioprocesses. [ABSTRACT FROM AUTHOR]