Synthesis of soybean soluble polysaccharide-based eco-friendly emulsions for soil erosion prevention and control.

This paper introduces the synthesis of an environmentally friendly emulsion that can be used as a soil anti-water erosion material. SSPS-g-P(BA-co-MMA-co-AA) emulsions were prepared using free radical copolymerization with soybean soluble polysaccharide (SSPS), acrylic acid (AA), butyl acrylate (BA), and methyl methacrylate (MMA). The structure, thermal stability, and morphology were characterized using FT-IR,TG,SEM, and particle diameter analysis. The resistance to water erosion, compressive strength and water retention of emulsion-treated loess/laterite was studied and germination tests were conducted. The results demonstrated that the duration of washout resistance of loess with 0.50 wt% emulsion exceeded 99 h, and the water erosion rate was 56.0 % after 72 h, while the water erosion rate of pure loess is 100.0 % after 4 min;the duration of washout resistance of laterite with 0.50 wt% emulsion exceeded 2 h, which was 8 times longer than pure laterite;The compressive strengths of 0.5 wt% emulsion-treated loess/laterite were 3.5 Mpa and 5.8 MPa, respectively, which were 7 and 9 times higher than that of pure soil. The plant seeds germinated normally half a month after planting. These findings suggest that emulsions can be used to control soil erosion without affecting the germination of plant seeds.

Establishment of a selectable marker recycling system for iterative gene editing in Fusarium fujikuroi.

Gibberellic acid (GA3) is a vital plant growth hormone widely used in agriculture. Currently, GA3 production relies on liquid fermentation by the filamentous fungus Fusarium fujikuroi. However, the lack of an effective selection marker recycling system hampers the application of metabolic engineering technology in F. fujikuroi, as multiple-gene editing and positive-strain screening still rely on a limited number of antibiotics. In this study, we developed a strategy using pyr4-blaster and CRISPR/Cas9 tools for recycling orotidine-5'-phosphate decarboxylase (Pyr4) selection markers. We demonstrated the effectiveness of this method for iterative gene integration and large gene-cluster deletion. We also successfully improved GA3 titers by overexpressing geranylgeranyl pyrophosphate synthase and truncated 3-hydroxy-3-methyl glutaryl coenzyme A reductase, which rewired the GA3 biosynthesis pathway. These results highlight the efficiency of our established system in recycling selection markers during iterative gene editing events. Moreover, the selection marker recycling system lays the foundation for further research on metabolic engineering for GA3 industrial production.

Species-specificity of the secondary biosynthetic potential in Bacillus.

Introduction: Although Bacillus species have produced a wide variety of structurally diverse and biologically active natural products, the secondary biosynthetic potential of Bacillus species is widely underestimated due to the limited number of biosynthetic gene clusters (BGCs) in this genus. The significant variation in the diversity and novelty of BGCs across different species within the Bacillus genus presents a major obstacle to the efficient discovery of novel natural products from Bacillus. Methods: In this study, the number of each class of BGCs in all 6,378 high-quality Bacillus genomes was predicted using antiSMASH, the species-specificity of BGC distribution in Bacillus was investigated by Principal component analysis. Then the structural diversity and novelty of the predicted secondary metabolites in Bacillus species with specific BGC distributions were analyzed using molecular networking. Results: Our results revealed a certain degree of species-specificity in the distribution of BGCs in Bacillus, which was mainly contributed by siderophore, type III polyketide synthase (T3PKS), and transAT-PKS BGCs. B. wiedmannii, B. thuringiensis, and B. cereus are rich in RiPP-like and siderophore BGCs, but lack T3PKS BGCs, while B. amyloliquefaciens and B. velezensis are abundant in transAT-PKS BGCs. These Bacillus species collectively encode 77,541 BGCs, with NRPS and RiPPs being the two most dominant types, which are further categorized into 4,291 GCFs. Remarkably, approximately 54.5% of GCFs and 93.8% of the predicted metabolite scaffolds are found exclusively in a single Bacillus species. Notably, B. cereus, B. thuringiensis, and B. velezensis exhibit the highest potential for producing species-specific NRPS and PKS bioinformatic natural products. Taking two species-specific NRPS gene clusters as examples, the potential of Bacillus to synthesize novel species-specific natural products is illustrated. Conclusion: This study highlights the species-specificity of the secondary biosynthetic potential in Bacillus and provides valuable insights for the targeted discovery of novel natural products from this genus.

Global analysis of the biosynthetic chemical space of marine prokaryotes.

BACKGROUND: Marine prokaryotes are a rich source of novel bioactive secondary metabolites for drug discovery. Recent genome mining studies have revealed their great potential to bio-synthesize novel secondary metabolites. However, the exact biosynthetic chemical space encoded by the marine prokaryotes has yet to be systematically evaluated. RESULTS: We first investigated the secondary metabolic potential of marine prokaryotes by analyzing the diversity and novelty of the biosynthetic gene clusters (BGCs) in 7541 prokaryotic genomes from cultivated and single cells, along with 26,363 newly assembled medium-to-high-quality genomes from marine environmental samples. To quantitatively evaluate the unexplored biosynthetic chemical space of marine prokaryotes, the clustering thresholds for constructing the biosynthetic gene cluster and molecular networks were optimized to reach a similar level of the chemical similarity between the gene cluster family (GCF)-encoded metabolites and molecular family (MF) scaffolds using the MIBiG database. The global genome mining analysis demonstrated that the predicted 70,011 BGCs were organized into 24,536 mostly new (99.5%) GCFs, while the reported marine prokaryotic natural products were only classified into 778 MFs at the optimized clustering thresholds. The number of MF scaffolds is only 3.2% of the number of GCF-encoded scaffolds, suggesting that at least 96.8% of the secondary metabolic potential in marine prokaryotes is untapped. The unexplored biosynthetic chemical space of marine prokaryotes was illustrated by the 88 potential novel antimicrobial peptides encoded by ribosomally synthesized and post-translationally modified peptide BGCs. Furthermore, a sea-water-derived Aquimarina strain was selected to illustrate the diverse biosynthetic chemical space through untargeted metabolomics and genomics approaches, which identified the potential biosynthetic pathways of a group of novel polyketides and two known compounds (didemnilactone B and macrolactin A 15-ketone). CONCLUSIONS: The present bioinformatics and cheminformatics analyses highlight the promising potential to explore the biosynthetic chemical diversity of marine prokaryotes and provide valuable knowledge for the targeted discovery and biosynthesis of novel marine prokaryotic natural products. Video Abstract.

Characterization of Secondary Metabolites from Mycelial Cultures of Black Morel Mushroom Morchella importuna (Ascomycota).

Species of the genus Morchella are highly prized worldwide for their excellent flavor and high medicinal value. In recent years, artificial cultivations of medicinal fungi with many advantages have elicited great interest as a promising alternative to produce certain valuable metabolites. Therefore, the secondary metabolites of fermented M. importuna belonging to the black morel clade isolated from China were investigated. The strain was cultured in a fermentation tank in PDB liquid medium by two-step method. The mycelia and fermentation broth were extracted by ethyl acetate. The secondary metabolites were separated and purified by repeated silica gel column chromatography. Structures of compounds were determined by NMR data and references. One new natural compound (1) and six known compounds (2-7) were obtained. Compounds 1, 2, 4, and 5 were first isolated from genus Morchella and compounds 3, 6, and 7 are first isolated from species M. importuna.

Safety and antifatigue effect of Korean Red Ginseng capsule: A randomized, double-blind and placebo-controlled clinical trial.

Background: In physical activity or labor, the human body is in a state of high intensity stress, and all parts or physiological functions of the body respond positively to maintain or balance the need for movement. The human body has many physiological changes in the process of movement, and fatigue is the external manifestation of various complex changes inside the human body. Fatigue is also a physiological mechanism of self-protection after the body reaches a certain level of activity, which can prevent the occurrence of life-threatening excessive functional failure. The generation of fatigue is a very complex process, and its mechanism has not been concluded yet. Therefore, it is an important work to search and screen the effective components of natural plants that have anti-fatigue effect and to explore their mechanism. Methods: This was a 8-week, randomized, double-blind, placebo-controlled clinical trial. A total of 110 subjects who passed physical examination were included according to the scheme design, and randomly divided into a test group which was given KRG and a placebo control group. The calculation is carried out according to the standard of sub-high-intensity exercise test. Results: There was no adverse effect on safety index of subjects after taking red ginseng capsule. After KRG treatment, subjective strength grade is significant lower than placebo treatment. Blood lactic acid content is significantly get lower after trial in KRG group, and significant lower than placebo group. Creatine phosphokinase(CK) content is significantly get lower after trial in KRG group, and significant lower than placebo group. Conclusion: According to the criterion in the test scheme, the result shows that KRG is helpful on relieving physical fatigue.

RETRACTED ARTICLE: Mutagenesis combined with fermentation optimization to enhance gibberellic acid GA3 yield in Fusarium fujikuroi.

Gibberellic acid (GA3) is a plant growth hormone that plays an important role in the production of crops, fruits, and vegetables with a wide market share. Due to intrinsic advantages, liquid fermentation of Fusarium fujikuroi has become the sole method for industrial GA3 production, but the broader application of GA3 is hindered by low titer. In this study, we combined atmospheric and room-temperature plasma (ARTP) with ketoconazole-based screening to obtain the mutant strain 3-6-1 with high yield of GA3. Subsequently, the medium composition and fermentation parameters were systematically optimized to increase the titer of GA3, resulting in a 2.5-fold increase compared with the titer obtained under the initial conditions. Finally, considering that the strain is prone to substrate inhibition and glucose repression, a new strategy of fed-batch fermentation was adopted to increase the titer of GA3 to 575.13 mg/L, which was 13.86% higher than the control. The strategy of random mutagenesis combined with selection and fermentation optimization developed in this study provides a basis for subsequent research on the industrial production of GA3.

Removal of multiple metal ions from wastewater by a multifunctional metal-organic-framework based trap.

The design and preparation of multifunctional adsorbent for practical wastewater treatment is still an enormous challenge. To remove multiple metal ions from wastewater, we developed a broad-spectrum metal ions trap named UIO-67-EDTA by incorporation of ethylenediaminetetraacetic acid into robust UIO-67. The adsorption experiments for 15 kinds of heavy metal ions including hard acid (Mn2+, Ba2+, Al3+, Cr3+, Fe3+), borderline acid (Co2+, Ni2+, Cu2+, Zn2+, Pb2+, Sn2+, Bi2+), soft acid (Ag+, Cd2+, Hg2+), and two kinds of dissolved minerals (Mg2+, Ca2+) show that the trap is very effective both in batch adsorption processes and breakthrough processes. At a pH value of 4.0, the removal efficiency for all metal ions was over 98% within 10 min, and the maximum static adsorption capacity for the representative metal ions Cr3+, Hg2+and Pb2+ was up to 416.67, 256.41, and 312.15 mg g-1, respectively. The adsorption kinetics fitted well with the pseudo-second-order model, indicating that the chemical adsorption was the rate-determining step in the adsorption process. Meanwhile, the material showed high stability and recyclability, the removal efficiency for the three representative metals was still maintained over 93% after five consecutive adsorption cycles.

Microbacterium karelineae sp. nov. isolated from a halophyte plant in the Taklamakan desert.

A bacterial strain, designated TRM 80801T, was isolated from the Karelinea in Taklamakan desert, Xinjiang Uygur Autonomous Region, north-west China. Cells were Gram-stain-positive, aerobic, non-motile, short rods. Strain TRM 80801T grew at 4-50 °C, with optimum growth at 28 °C, and grew at pH 6.0-11.0 and 1-15 % (w/v) NaCl. Phylogenetic analyses of the 16S rRNA gene sequences placed strain TRM 80801T within the genus Microbacterium with the highest similarities to Microbacterium suaedae YZYP 306T (98.97 %) and Microbacterium indicum BBH6T (98.17 %), respectively. The DNA G+C content of TRM 80801T is 69.38 mol%. The cell-wall peptidoglycan contained the amino acids ornithine, glutamic acid, glycine and alanine, the diagnostic diamino acid was ornithine. The acyl type of the peptidoglycan was glycolyl. Whole-cell sugars were ribose, mannose, glucose, rhamnose and galactose. The major cellular fatty acids were anteiso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0. The predominant menaquinones were MK-10, MK-11 and MK-12. The polar lipids were diphosphatidylglycerol, phosphatidylglycerol. The whole-genome average nucleotide identity (ANI) value between strain TRM 80801T and Microbacterium suaedae YZYP 306T is 70.2 %. On the basis of the evidence presented in this study, strain TRM 80801T is representative of a novel species in the genus Microbacterium, for which the name Microbacterium karelineae sp. nov. is proposed. The type strain is TRM 80801T (=CCTCC AB 2019248T=KCTC 49357T).

YALIcloneNHEJ: An Efficient Modular Cloning Toolkit for NHEJ Integration of Multigene Pathway and Terpenoid Production in Yarrowia lipolytica.

Non-homologous end-joining (NHEJ)-mediated random integration in Yarrowia lipolytica has been demonstrated to be an effective strategy for screening hyperproducer strains. However, there was no multigene assembly method applied for NHEJ integration, which made it challenging to construct and integrate metabolic pathways. In this study, a Golden Gate modular cloning system (YALIcloneNHEJ) was established to develop a robust DNA assembly platform in Y. lipolytica. By optimizing key factors, including the amounts of ligase and the reaction cycles, the assembly efficiency of 4, 7, and 10 fragments reached up to 90, 75, and 50%, respectively. This YALIcloneNHEJ system was subsequently applied for the overproduction of the sesquiterpene (-)-α-bisabolol by constructing a biosynthesis route and enhancing the flux in the mevalonate pathway. The resulting strain produced 4.4 g/L (-)-α-bisabolol, the highest titer reported in yeast to date. Our study expands the toolbox of metabolic engineering and is expected to enable a highly efficient production of various terpenoids.

Investigation of Chemical Compounds and DPPH Radical Scavenging Activity of Oudemansiella raphanipes (Agaricomycetes) Based on Fermentation.

Oudemansiella raphanipes, an edible medicinal mushroom, is very popular in the market. With the development of technology, liquid fermentation is becoming more and more popular in edible fungi cultivation. Strain O. raphanipes was cultured by submerged fermentation and secondary metabolites were investigated simultaneously. Six chemical constituents including two unsaturated fatty acids (1-2), one ergosterol (3), one phenol (4), and two benzamides (5-6) were acquired from ethyl acetate extract. Their structures were mainly elucidated by NMR spectral data and known products. All compounds were isolated from O. raphanipes for the first time. DPPH radical scavenging activity was evaluated. Orcinol (4) exhibited remarkable antioxidant activity; its free radical scavenging rate can reach up to 80% of 20 mg/mL. 2-pyruvoybenzamide (5) and 2-(2-hydroxypropanamido) benzamide (6) showed less active antioxidant capacity.

Preparation of Uracil by bacteria isolated from Morchella.

Morchella is one of the most famous rare edible and medicinal fungi over the world. Highly nutritious and immature cultivation techniques led to the high price and the markets have remained tight. The pathogenic bacteria were serious in artificial cultivation of Morchella that affected the growth and yield of Morchella. Isolation of pathogenic bacteria and metabolites were investigated in order to improve the artificial cultivation technology. The isolated strain (YDJZ-01-01C) was identified by Gram staining and sequence of 16S rDNA. Structures of metabolites were confirmed based on NMR spectra and literatures. However, the main products were uracil and thymine that considered as important intermediate of anti-tumor 5-fluorouracil. Interestingly, a new synthetic pathway for preparation of uracil by microorganism was found except for chemical synthesis. The new preparation pathway provided mild, green, sustainable and environment friendly method to produce uracil that meets the needs of modern chemistry.

Chemical constituents of fungus F03 belonging to Basidiomycota.

Filamentous fungus F03 belonging to Basidiomycota was obtained and identified as Phlebiopsis crassa based on ITS sequence when Morchella. sp was isolated from the wild fruiting body by spores releasing method. Chemical constituents were separated by gel chromatography, HPLC and recrystallization. Structures of compounds were confirmed by NMR data. Four products orsellinic acid (1), α-nigerose (2), uridine (3), N-(4-hydroxyphenyl)acetamide (4) were identified and all compounds were isolated from the genus Phlebiopsis for the first time.

Secondary Metabolites and Antiradical Activity of Liquid Fermentation of Morchella sp. Isolated from Southwest China.

Morels famous for their taste and nutrition are in short supply all over the world although they were considered as one of the most highly prized edible and medicinal mushrooms. Because of the limitation of resource and cultivation technology, fermentation of edible mushroom was gradually applied to nutrient, bioactivity and breeder seed preparation. At present, there are more reports on sugar and amino acid but less on other components. Morchella sp. YDJ-ZY-1 was isolated from the wild fruiting body by the spores releasing method in Zunyi Guizhou province in Southwest China and identified based on phenotype and genotype characteristics. Chemical compositions of YDJ-ZY-1 were investigated from liquid fermentation that will lay the foundation for further development and utilization. Four pyranoids (1-4) and 2-(1-oxo-2-hydroxyethyl) furan (5), linoleic acid (6), Morelin (2-hydroxy-cinnamic acid methyl ester, (7) and 1-O-ß-d-ribofuranose-Morelin (8) were obtained from EtOAc extraction and elucidated by spectral data. Product 4 and 8 were new compounds and 7 was isolated from nature for the first time. Antiradical activity was evaluated by free radical scavenging effect on DPPH (1,1-Diphenyl-2-picrylhydrazyl radical 2,2-Diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl). Compound 5 exhibited strong antiradical activity while compounds 1 and 2 exhibited moderate activity. Thus, incubation of Morchella sp YDJ-ZY-1 separated from the wild fruit body afforded eight compounds. Secondary metabolites with new structures were mined from fermentation of Morchella sp. and antiradical activity was evaluated.

New pyrazinoquinazoline alkaloids Isolated from a culture of Stenotrophomonas maltophilia QB-77.

Two new pyrazinoquinazoline alkaloids, epi-fiscalin D (1) and epi-fiscalin E (2), as well as three known analogues, norquinadoline A (3), quinadoline A (4), and fiscalin C (5), were isolated from ethyl acetate extract of the fermentation broth of Stentrophomonas maltophilia QB-77. The structures of new compounds were elucidated on the basis of extensive spectroscopic data analysis including UV, HRESIMS, and 1D and 2D NMR experiments. All the isolated compounds were tested for their in vitro cytotoxicity against five human cancer cell lines (SMMC-7721, MCF-7, HL-60, SW480, and A-549) and antibacterial activities against Bacillus subtilis, Escherichia coli, and Staphylococcus aureus.

A simple ratiometric fluorescent sensor for fructose based on complexation of 10-hydroxybenzo[h]quinoline with boronic acid.

A simple ratiometric fluorescent sensor for fructose was presented. It consisted of 10-hydroxybenzo[h]quinoline (HBQ) which showed emission at 572nm and 3-pyridylboronic acid (PDBA) whose complex with HBQ gave emission at 500nm. The reaction of fructose with PDBA inhibited the complexation of HBQ with PDBA, resulting in the change of dual-emission intensity ratio. The sensor well quantified fructose in the range of 0.015-2.5mM with detection limit of 0.005mM. Besides, this sensor exhibited excellent selectivity and was successfully applied to fructose detection in food. This work provides a simple ratiometric sensing platform for sensitive and selective detection of fructose.

Sulindac attenuates valproic acid-induced oxidative stress levels in primary cultured cortical neurons and ameliorates repetitive/stereotypic-like movement disorders in Wistar rats prenatally exposed to valproic acid.

Accumulating evidence suggests that anti-inflammatory agents and antioxidants have neuroprotective properties and may be beneficial in the treatment of neurodevelopental disorders, such as autism. In the present study, the possible neuroprotective properties of sulindac, a non-steroidal anti-inflammatory drug (NSAID), were investigated in vitro using cultured cortical neurons with valproic acid (VPA)-induced neurotoxicity, as well as in vivo through the behavioral analysis of rats prenatally exposed to VPA as a model of autism. VPA induced 4-hydroxynonenal (4-HNE) expression, reactive oxygen species (ROS) generation and decreased cell viability in primary cultured cortical neurons established from timed-pregnant (embryonic day 18) Wistar rat pups. However, co-incubation of the neurons with VPA and sulindac reduced oxidative stress and increased cell viability. The rats were administered an intraperitoneal injection with one of the following: VPA, sulindac, VPA and sulindac, or physiological saline, and their offspring were subjected to the open field test. During the test trials, repetitive/stereotypic-like movements for each rat were recorded and analyzed. The results revealed that treatment with both sulindac and VPA reduced the VPA-induced repetitive/stereotypic-like activity and the sulindac and VPA-treated animals responded better in the open field test compared to the VPA-treated animals. The results from the present study demonstrate that the antioxidant properties of sulindac may prove to be beneficial in the treatment of autism, suggesting that the upregulation of the Wnt/ß-catenin signaling pathway disrupts oxidative homeostasis and facilitates susceptibility to autism.

Induction of Tca8113 tumor cell apoptosis by icotinib is associated with reactive oxygen species mediated p38-MAPK activation.

Icotinib, a selective EGFR tyrosine kinase inhibitor (EGFR-TKI), has been shown to exhibit anti-tumor activity against several tumor cell lines. However, the exact molecular mechanism of icotinib's anti-tumor effect remains unknown. This study aims to examine the zytotoxic effect of icotinib on Tca8113 cells and its potential molecular mechanism. Icotinib significantly resulted in dose-dependent cell death as determined by MTT assay, accompanied by increased levels of Bax and DNA fragmentation. Icotinib could also induce Reactive Oxygen Species (ROS) generation. Further studies confirmed that scavenging of reactive oxygen species by N-acetyl-L-cysteine (NAC), and pharmacological inhibition of MAPK reversed icotinib-induced apoptosis in Tca8113 cells. Our data provide evidence that icotinib induces apoptosis, possibly via ROS-mediated MAPK pathway in Tca8113 cells.

Icotinib inhibits the invasion of Tca8113 cells via downregulation of nuclear factor κB-mediated matrix metalloproteinase expression.

Icotinib is an epidermal growth factor receptor tyrosine kinase inhibitor, which has been revealed to inhibit proliferation in tumor cells. However, the effect of icotinib on cancer cell metastasis remains to be explained. This study examines the effect of icotinib on the migration and invasion of squamous cells of tongue carcinoma (Tca8113 cells) in vitro. The results of the Boyden chamber invasion assay demonstrated that icotinib reduced cell invasion, suppressed the protein levels of matrix metalloproteinases (MMPs), MMP-2 and MMP-9, and increased the expression of tissue inhibitor of metalloproteinase-1. In addition, icotinib was found to significantly decrease the protein levels of nuclear factor κB (NF-κB) p65, which suggested that icotinib inhibits NF-κB activity. Furthermore, treatment with the NF-κB inhibitor, pyrrolidine dithiocarbamate, suppressed cell invasion and MMP-2 expression. These results suggested that icotinib inhibits the invasion of Tca8113 cells by downregulating MMP via the inactivation of the NF-κB signaling pathways.