4-Coumarate-CoA ligase (4-CL) enhances flavonoid accumulation, lignin synthesis, and fruiting body formation in Ganoderma lucidum.

It is now understood that 4-Coumarate-CoA ligases (4-CL) are pivotal in bridging the phenylpropanoid metabolic pathway and the lignin biosynthesis pathway in plants. However, limited information on 4-CL genes and their functions in fungi is available. In this study, we cloned the 4-CL gene (Gl21040) from Ganoderma lucidum, which spans 2178 bp and consists of 10 exons and 9 introns. We also developed RNA interference and overexpression vectors for Gl21040 to investigate its roles in G. lucidum. Our findings indicated that in the Gl21040 interference transformants, 4-CL enzyme activities decreased by 31 %-57 %, flavonoids contents decreased by 10 %-22 %, lignin contents decreased by 20 %-36 % compared to the wild-type (WT) strain. Conversely, in the Gl21040 overexpression transformants, 4-CL enzyme activity increased by 108 %-143 %, flavonoids contents increased by 8 %-37 %, lignin contents improved by 15 %-17 % compared to the WT strain. Furthermore, primordia formation was delayed by approximately 10 days in the Gl21040-interferenced transformants but occurred 3 days earlier in the Gl21040-overexpressed transformants compared to the WT strain. These results underscored the involvement of the Gl21040 gene in flavonoid synthesis, lignin synthesis, and fruiting body formation in G. lucidum.

The transcription factor Ste12-like increases the mycelial abiotic stress tolerance and regulates the fruiting body development of Flammulina filiformis.

Introduction: Flammulina filiformis is one of the most commercially important edible fungi worldwide, with its nutritional value and medicinal properties. It becomes a good model species to study the tolerance of abiotic stress during mycelia growth in edible mushroom cultivation. Transcription factor Ste12 has been reported to be involved in the regulation of stress tolerance and sexual reproduction in fungi. Methods: In this study, identification and phylogenetic analysis of ste12-like was performed by bioinformatics methods. Four ste12-like overexpression transformants of F. filiformis were constructed by Agrobacterium tumefaciens-mediated transformation. Results and Discussion: Phylogenetic analysis showed that Ste12-like contained conserved amino acid sequences. All the overexpression transformants were more tolerant to salt stress, cold stress and oxidative stress than wild-type strains. In the fruiting experiment, the number of fruiting bodies of overexpression transformants increased compared with wild-type strains, but the growth rate of stipes slowed down. It suggested that gene ste12-like was involved in the regulation of abiotic stress tolerance and fruiting body development in F. filiformis.

Controllable Synthesis of Sheet-Flower ZnO for Low Temperature NO2 Sensor.

ZnO is a wide band gap semiconductor metal oxide that not only has excellent electrical properties but also shows excellent gas-sensitive properties and is a promising material for the development of NO2 sensors. However, the current ZnO-based gas sensors usually operate at high temperatures, which greatly increases the energy consumption of the sensors and is not conducive to practical applications. Therefore, there is a need to improve the gas sensitivity and practicality of ZnO-based gas sensors. In this study, three-dimensional sheet-flower ZnO was successfully synthesized at 60 °C by a simple water bath method and modulated by different malic acid concentrations. The phase formation, surface morphology, and elemental composition of the prepared samples were studied by various characterization techniques. The gas sensor based on sheet-flower ZnO has a high response value to NO2 without any modification. The optimal operating temperature is 125 °C, and the response value to 1 ppm NO2 is 125. At the same time, the sensor also has a lower detection limit (100 ppb), good selectivity, and good stability, showing excellent sensing performance. In the future, water bath-based methods are expected to prepare other metal oxide materials with unique structures.

Conversion of Waste Oil from Oil Refinery into Emulsion Liquid Membrane for Removal of Phenol: Stability Evaluation, Modeling and Optimization.

The waste oil emulsion liquid membrane produced by waste oil from oil refineries (WELM) is used to separate the phenol in purified water from the sour water stripper in oil refinery facilities, and the stability of WELM was studied. It is verified that waste refinery oil can be produced into emulsion liquid membrane with good stability and high removal rate for the first time. The WELM stability models were established by response surface methodology (RSM) and artificial neural network (ANN), respectively. The principle and mechanism of various parameters, as well as the interaction effects on the stability of WELM, are proposed. The effects of parameters, including the ratio of Span-80, liquid paraffin, the ratio of internal and oil, and the rotational speed of the homogenizer, were investigated. Under the optimal operating parameters, the WELM had a demulsification percentage of just 0.481%, and the prediction results of RSM and ANN were 0.536% and 0.545%, respectively. Both models demonstrate good predictability. The WELM stability model has a high application value in the treatment of phenol-containing wastewater in the oil refining industry, and provides a green method of resource recovery.

A Mechanistic Study of Asymmetric Transfer Hydrogenation of Imines on a Chiral Phosphoric Acid Derived Indium Metal-Organic Framework.

A density functional theory (DFT) study is reported to examine the asymmetric transfer hydrogenation (ATH) of imines catalyzed by an indium metal-organic framework (In-MOF) derived from a chiral phosphoric acid (CPA). It is revealed that the imine and reducing agent (i.e., thiazoline) are simultaneously adsorbed on the CPA through H-bonding to form an intermediate, subsequently, a proton is transferred from thiazoline to imine. The transition state TS-R and TS-S are stabilized on the CPA via H-bonding. Compared to the TS-S, the TS-R has shorter H-bonding distances and longer C-H···π distances, it is more stable and experiences less steric hindrance. Consequently, the TS-R exhibits a lower activation barrier affording to the (R)-enantiomer within 68.1% ee in toluene. Imines with substituted groups such as -NO2, -F, and -OCH3 are used to investigate the substitution effects on the ATH. In the presence of an electron-withdrawing group like -NO2, the electrophilicity of imine is enhanced and the activation barrier is decreased. The non-covalent interactions and activation-strain model (ASM) analysis reveal that the structural distortions and the differential noncovalent interactions of TSs in a rigid In-MOF provide the inherent driving force for enantioselectivity. For -OCH3 substituted imine, the TS-S has the strongest steric hindrance, leading to the highest enantioselectivity. When the solvent is changed from toluene to dichloromethane, acetonitrile, and dimethylsulfoxide with increasing polarity, the activation energies of transition state increase whereas their difference decreases. This implies the reaction is slowed down and the enantioselectivity becomes lower in a solvent of smaller polarity. Among the four solvents, toluene turns out to be the best for the ATH. The calculated results in this study are in fairly good agreement with experimental observations. This study provides a mechanistic understanding of the reaction mechanism, as well as substitution and solvent effects on the activity and enantioselectivity of the ATH. The microscopic insights are useful for the development of new chiral MOFs toward important asymmetric reactions.

Transcriptome and metabolome analyses reveal transcription factors regulating ganoderic acid biosynthesis in Ganoderma lucidum development.

Ganoderma lucidum is an important medicinal fungus in Asian countries. Ganoderic acid (GA) is the major variety of bioactive and medicative components in G. lucidum. Biosynthesis of secondary metabolites is usually associated with cell differentiation and development. However, the mechanism underlying these phenomena remain unclear. Transcription factors play an essential regulatory role in the signal transduction pathway, owing to the fact that they represent the major link between signal transduction and expression of target genes. In the present study, we performed transcriptome and metabolome analyses to identify transcription factors involved in GA biosynthesis during development of G. lucidum. Transcriptome data revealed differentially expressed genes between mycelia and primordia, as well as between mycelia and the fruiting body. Results from gene ontology enrichment analysis and metabolome analyses suggested that GAs and flavonoids biosynthetic process significantly changed during fungal development. The analysis of predicted occurrences of DNA-binding domains revealed a set of 53 potential transcription factor families in G. lucidum. Notably, we found homeobox transcription factor and velvet family protein played important role in GA biosynthesis. Combined with previous studies, we provided a model diagram of transcription factors involved in GA biosynthesis during fruiting body formation. Collectively, these results are expected to enhance our understanding into the mechanisms underlying secondary metabolite biosynthesis and development in fungi.

Integrated Transcriptomics and Nontargeted Metabolomics Analysis Reveal Key Metabolic Pathways in Ganoderma lucidum in Response to Ethylene.

Ganoderic acid (GA) is an important secondary metabolite of Ganoderma lucidum with a diverse array of pharmacological properties. In this study, we found that exogenous ethylene increased the production of endogenous ethylene and ganoderic acid in G. lucidum. However, the mechanism by which ethylene is regulated remains unclear. As a result, we performed a combined transcriptomics and nontargeted metabolomics analysis to evaluate the regulatory mechanism of ethylene. A total of 4070 differentially expressed genes (1835 up-regulated and 2235 down-regulated) and 378 differentially accumulated metabolites (289 up-regulated and 89 down-regulated) were identified in all groups. The transcriptomics and nontargeted metabolomics data revealed that genes involved in the tricarboxylic acid (TCA) cycle, polyamine metabolic pathway, acetyl-CoA carboxylase (ACC) pathway, and triterpenoid metabolism were up-regulated, whereas the metabolic intermediates involved in these metabolic pathways were down-regulated. These findings imply that ethylene potentially accelerates normal glucose metabolism, hence increasing the number of intermediates available for downstream biological processes, including polyamine metabolism, ethylene synthesis pathway, and ganoderic acid biosynthesis. The findings will contribute significantly to our understanding of secondary metabolites biosynthesis in fungi.

Transcriptomic and Non-Targeted Metabolomic Analyses Reveal the Flavonoid Biosynthesis Pathway in Auricularia cornea.

Flavonoids, which are abundant in plants, are recognized for their antioxidant and anticancer roles in clinical applications. However, little is known about the molecular basis of flavonoid biosynthesis in fungi. In this study, we found that inclusion of leachate of Korshinsk peashrub (Caragana korshinskii) in the fermentation medium increased the total flavonoid content of the edible fungus Auricularia cornea by 23.6% relative to that grown in a control medium. Combined transcriptomic and non-targeted metabolomic analysis of the flavonoid biosynthesis pathway in A. cornea illustrated that there are important metabolites in the phenylpropanoid, coumarin and isoflavonoid biosynthesis pathways. In addition, we found that certain homologous genes encode phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO) and chalcone isomerase (CHI) in these biosynthesis pathways. These results, in this study, provide a new line for studying the regulation of flavonoid production in edible fungi.

Preparation and Separation Properties of Electrospinning Modified Membrane with Ionic Liquid Terminating Polyimide/Polyvinylpyrrolidone@Polydopamine.

In this paper, superhydrophilic polyimide (PI) membranes were prepared using the electrostatic spinning method, capped with a hydrophilic ionic liquid (IL), and blended with polyvinylpyrrolidone (PVP). Using this preparation, the surface of the fiber membranes was coated in polydopamine (PDA) by means of an in-growth method. Scanning electron micrographs showed prepared blend films can form continuous fibers, for whom the distributions of diameter and pore were uniform. Post-modification (carried out by adding hydrophilic substances), the ability of the membrane surface to adhere to water was also significantly improved. The water contact angle was reduced from 128.97 ± 3.86° in unmodified PI to 30.26 ± 2.16°. In addition, they displayed a good separation effect on emulsified oil/water mixtures. The membrane flux reached a maximum value of 290 L·m-2·h-1, with a maximum separation efficiency reached of more than 99%. After being recycled 10 times, the separation efficiency maintained a level exceeding 95%. The purpose of this study is to demonstrate the simplicity and efficiency of this experiment, thereby providing new ideas for the future application of membrane separation technology in wastewater treatment.

Highly Selective Gas Sensor Based on Litchi-like g-C3N4/In2O3 for Rapid Detection of H2.

Hydrogen (H2) has gradually become a substitute for traditional energy, but its potential danger cannot be ignored. In this study, litchi-like g-C3N4/In2O3 composites were synthesized by a hydrothermal method and used to develop H2 sensors. The morphology characteristics and chemical composition of the samples were characterized to analyze the gas-sensing properties. Meanwhile, a series of sensors were tested to evaluate the gas-sensing performance. Among these sensors, the sensor based on the 3 wt% g-C3N4/In2O3 (the mass ratio of g-C3N4 to In2O3 is 3:100) showeds good response properties to H2, exhibiting fast response/recovery time and excellent selectivity to H2. The improvement in the gas-sensing performance may be related to the special morphology, the oxygen state and the g-C3N4/In2O3 heterojunction. To sum up, a sensor based on 3 wt% g-C3N4/In2O3 exhibits preeminent performance for H2 with high sensitivity, fast response, and excellent selectivity.

The Fvclp1 gene regulates mycelial growth and fruiting body development in edible mushroom Flammulina velutipes.

Fruiting body development in Agaricomycetes represents the most complex and unclear process in the fungi. Mating type pathways (A and B) and transcription factors are important regulators in the sexual development of mushrooms. It is known that clampless1 (clp1) is an additional gene that participate under the homeodomain (HD) genes in the matA pathway and clp1 inactivation blocks clamps formation in Coprinopsis cinerea. In this study we identified and analyzed a homologous Fvclp1 gene in the edible mushroom Flammulina velutipes. The coding sequence of the Fvclp1 was 1011 bp without intron interruption, encoding a protein of 336 amino acids. To exhibit the role of Fvclp1 in clamp development and fruiting body formation, knockdown and overexpression mutants were prepared. No significant difference was observed in the monokaryotic hyphal morphology of overexpression and knockdown transformants. In the dikaryotic hyphae from the compatible crossings between the wild-type L22 strain and Fvclp1 knockdown or overexpression mutants, clamp connections developed. However, knockdown mutants could generate fewer fruiting bodies than the wild-type strain. On the contrary, reduced mycelial growth rate but improved fruiting ability was observed in the dikaryotic Fvclp1 overexpression mutants as compared to the wild-type strain. These results indicate that Fvclp1 is necessary and actively involved in fruiting body development in F. velutipes. Overall, these findings suggest that further studies on the function of Fvclp1 would advance our understanding of sexual reproduction and fruiting body development in edible mushrooms.

The transcription factor FvHmg1 negatively regulates fruiting body development in Winter Mushroom Flammulina velutipes.

Fruiting body formation in Agaricomycetes represents the most complex and unclear process in the fungi. Mating type pathways (matA and matB) and transcription factors are important regulators in the process. Here, we report a new High-mobility-group (HMG) box domain protein FvHmg1 that acts as a negative transcription regulator in fruiting body development in Winter Mushroom Flammulina velutipes. However, the expression of Fvhmg1 in dikaryon and primordial stages was significantly lower than that of monokaryon. The Fvhmg1-RNAi mutants had a better ability of fruiting than wild type strain. Overall expression of Fvhmg1 was controlled under compatible matA and matB genes where compatible matA genes could increase its expression level, while compatible matB genes had the opposite effect. It means when two monokaryons with compatible matA and matB genes were crossed, the negatively transcription factor FvHmg1 was inhibited, and normal fully fruiting body could formation and develop. The relationship between FvHmg1 and mating type pathway would advance to understand of sexual reproduction and fruiting body development in edible mushrooms.

Study of thermotolerant mechanism of Stropharia rugosoannulata under high temperature stress based on the transcriptome sequencing.

Stropharia rugosoannulata is a popular edible mushroom in the world. High temperature seriously affects its yield and quality. In this study, transcriptome sequencing was performed on the mycelia of heat-resistant strains and heat-sensitive strains heat-treated at 38 °C for 0 h and 24 h. The changes of catalase (CAT) activity, superoxide dismutase (SOD) activity and trehalose content in the mycelia under high temperature stress were also measured and analyzed. We find that the differential genes are mainly enriched in the pathways of glycerophospholipid metabolism, starch and sucrose metabolism, protein processing in the endoplasmic reticulum, etc. The expression levels of genes encoding trehalose-6-phosphate phosphatase (TPP), CAT, SOD, etc. are quite different. And these genes' variation range in the thermotolerant strain are higher than that in heat-sensitive strain. The CAT activity and trehalose content of the two strains increase first and then decrease, and the SOD activity increase slowly. The CAT, SOD activity and trehalose content of the thermotolerant strain are higher than those of the heat-sensitive strain. This study will provide a basis for further research on important signal pathways and gene function identification of S. rugosoannulata related to high temperature stress.

Optimization of a MOF Blended with Modified Polyimide Membrane for High-Performance Gas Separation.

The preparation, characterization and gas separation properties of mixed matrix membranes (MMMs) were obtained from polyimide capped with ionic liquid and blended with metal-organic frameworks (MOFs). The synthesized MOF was amine functionalized to produce UiO-66-NH2, and its amino group has a higher affinity for CO2. Mixed matrix membranes exhibited good membrane forming ability, heat resistance and mechanical properties. The polyimide membrane exclusively capped by ionic liquid exhibited good permselectivity of 74.1 for CO2/CH4, which was 6.2 times that of the pure polyimide membrane. It is worth noting that MMM blended with UiO-66-NH2 demonstrated the highest ideal selectivity for CO2/CH4 (95.1) with a CO2 permeability of 7.61 Barrer, which is close to the 2008 Robeson upper bound. The addition of UiO-66-NH2 and ionic liquid enhanced the permselectivity of MMMs, which may be one of the promising technologies for high performance CO2/CH4 gas separation.

A New Environmentally-Friendly System for Extracting Cellulose from Corn Straw: The Low Temperature Laccase System.

Corn straw is an agricultural waste. The system for extracting cellulose from corn straw at a high temperature has been widely reported by researchers. However, the system for extracting cellulose from corn straw at a low temperature has been rarely reported. In this paper, a new system for extracting cellulose from corn straw at a low temperature was reported for the first time. This new system is designated as the low temperature laccase system (LTLS). Cellulose was successfully extracted from corn straw by the LTLS, and the used solution could be recycled. Therefore, the low temperature laccase system is an environmentally-friendly system. The cellulose content in corn straw is 30-40%. The yield of cellulose extracted by LTLS was 33%. The obtained cellulose product was creamy white. The extracted cellulose samples were characterized by using infrared spectroscopy (IR), thermogravimetry (TG), and X-ray diffraction (XRD). The results were consistent with that of standard cellulose. We confirmed that the LTLS extracted cellulose from corn straw with high purity.

High Efficiency Gas Permeability Membranes from Ethyl Cellulose Grafted with Ionic Liquids.

Ethyl cellulose was grafted with ionic liquids in optimal yields (62.5-64.1%) and grafting degrees (5.93-7.90%) by the esterification of the hydroxyl groups in ethyl cellulose with the carboxyl groups in ionic liquids. In IR spectra of the ethyl cellulose derivatives exhibited C=O bond stretching vibration peaks at 1760 or 1740 cm-1, confirming the formation of the ester groups and furnishing the evidence of the successful grafting of ethyl cellulose with ionic liquids. The ethyl cellulose grafted with ionic liquids could be formed into membranes by using the casting solution method. The resulting membranes exhibited good membrane forming ability and mechanical properties. The EC grafted with ionic liquids-based membranes demonstrated PCO2/PCH4 separation factors of up to 18.8, whereas the PCO2/PCH4 separation factor of 9.0 was obtained for pure EC membrane (both for CO2/CH4 mixture gas). The membranes also demonstrated an excellent gas permeability coefficient PCO2, up to 199 Barrer, which was higher than pure EC (PCO2 = 46.8 Barrer). Therefore, it can be concluded that the ionic liquids with imidazole groups are immensely useful for improving the gas separation performances of EC membranes.

Hydrothermal synthesis of hierarchical CoO/SnO2 nanostructures for ethanol gas sensor.

In this work, ethanol gas sensor with high performance was fabricated successfully with hierarchical CoO/SnO2 heterojunction by two-steps hydrothermal method. The response value of CoO/SnO2 sensor is up to 145 at 250 °C when exposed to 100 ppm ethanol gas, which is much higher than that (13.5) of SnO2 sensor. These good sensing performances mainly attribute to the formation of the CoO/SnO2 heterojunction, which makes great variation of resistance in air and ethanol gas. Thus, the combination of n-type SnO2 and p-type CoO provides an effective strategy to design new ethanol gas sensors. The unique nanostructure also played an important role in detecting ethanol, due to its contribution in facilitating the transport rate of the ethanol gas molecules. Also, we provide a general two-step strategy for designing the heterojunction based on the SnO2 nanostructure.

The Morphologies of the Semiconductor Oxides and Their Gas-Sensing Properties.

Semiconductor oxide chemoresistive gas sensors are widely used for detecting deleterious gases due to low cost, simple preparation, rapid response and high sensitivity. The performance of gas sensor is greatly affected by the morphology of the semiconductor oxide. There are many semiconductor oxide morphologies, including zero-dimensional, one-dimensional, two-dimensional and three-dimensional ones. The semiconductor oxides with different morphologies significantly enhance the gas-sensing performance. Among the various morphologies, hollow nanostructures and core-shell nanostructures are always the focus of research in the field of gas sensors due to their distinctive structural characteristics and superior performance. Herein the morphologies of semiconductor oxides and their gas-sensing properties are reviewed. This review also proposes a potential strategy for the enhancement of gas-sensing performance in the future.

Elucidation of a carboxylate O-methyltransferase NcmP in nocamycin biosynthetic pathway.

Nocamycins belong to the tetramic acid family natural products and show potent antimicrobial activity. Recently, the biosynthetic gene cluster of nocamycin was identified from the rare actinomycete Saccharothrix syringae and an S-adenosylmethionine (SAM) dependent methyltransferase gene NcmP was found to be located within the gene cluster. In this report, the methyltransferase gene NcmP was disrupted and a new nocamycin intermediate nocamycin E was isolated from the mutant strain. Meanwhile, NcmP was heterologously expressed in Escherichia coli BL21 (DE3) and biochemically characterized as a carboxylate O-methyltransferase in nocamycin biosynthetic pathway. Compared to nocamycin I, nocamycin E showed inferior antibacterial activity, indicating the methyl group is essential to antibacterial activity.

Porous α-Fe2O3 microflowers: Synthesis, structure, and enhanced acetone sensing performances.

Porous α-Fe2O3 microflowers, which were composed of many nanospindles assembled by large numbers of nanoparticles, were successfully synthesized by calcining the FeSO4(OH) precursor prepared through a simple ethanol-mediated method. Various techniques were employed to obtain the crystalline and morphological properties of the as-prepared products. The formation process of such microstructure was proposed according to the morphology and component of the products obtained at different reaction time. Moreover, the obtained α-Fe2O3 was utilized as sensing materials upon exposure to various test gases. As expected, in virtue of the less-agglomerated configuration and unique porous structure, the hierarchical α-Fe2O3 microflowers exhibited higher response as well as faster response/recovery time to acetone when compared with α-Fe2O3 nanoparticles. Significantly, the response time was measured to be 1s at the low operating temperature of 210°C.