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165 result(s) for "Hirai, Hirofumi"
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Transcriptomic analysis reveals ligninolytic enzymes of white-rot fungus Phanerochaete sordida YK-624 participating in bisphenol F biodegradation under ligninolytic conditions
Bisphenol F (BPF) is widely used in the plastic manufacturing industry as a replacement for bisphenol A (BPA) because BPF and BPA have similar structures and comparable properties. However, BPF is ubiquitously present in the environment and has higher toxicity to humans. This study is the first to report BPF degradation using the white-rot fungus Phanerochaete sordida YK-624 under ligninolytic conditions (pH=4.5, 30 °C). P. sordida YK-624 almost completely degraded BPF within 4 days. Moreover, functional genes involved in BPF degradation were detected by RNA-Seq. Metabolic processes and peroxidases were enriched by GO analysis, and the metabolic pathway was enriched according to the KEGG pathway analysis. These results suggested that P. sordida YK-624 could secrete higher levels of ligninolytic enzymes lignin peroxidase (LiP) and manganese peroxidase (MnP) for BPF degradation. The results indicated that LiPs and MnPs are important for BPF degradation and cytochrome P450s play a small role. Furthermore, reliability of the RNA-Seq results was validated by qRT-PCR.
The complete mitochondrial genome of the white-rot fungus Phanerochaete sordida YK-624
The white-rot fungus Phanerochaete sordida (Karsten) Eriksson and Ryvarden 1978 is known for its excellent ligninolytic activity and capability to degrade various recalcitrant organic pollutants. In this study, we determined the complete mitochondrial genome sequence of P. sordida YK-624. The mitochondrial genome is 129,567 bp in length with a GC content of 28.9%, and contains two ribosomal RNA genes, 26 transfer RNA genes, and 50 open reading frames, including 14 conserved proteins. Phylogenetic analysis based on the mitochondrial genome confirmed that P. sordida belongs to the family Phanerochaetaceae in the order Polyporales, and showed the general phylogenetic relationships.
Axl, Immune Checkpoint Molecules and HIF Inhibitors from the Culture Broth of Lepista luscina
Two compounds 1 and 2 were isolated from the culture broth of Lepista luscina. This is the first time that compound 1 was isolated from a natural source. The structure of compound 1 was identified via 1D and 2D NMR and HRESIMS data. Compounds 1 and 2 along with 8-nitrotryptanthrin (4) were evaluated for their biological activities using the A549 lung cancer cell line. As a result, 1 and 2 inhibited the expression of Axl and immune checkpoint molecules. In addition, compounds 1, 2 and 4 were tested for HIF inhibitory activity. Compound 2 demonstrated statistically significant HIF inhibitory effects on NIH3T3 cells and 1 and 2 against ARPE19 cells.
Ergosterol and Its Metabolites Induce Ligninolytic Activity in the Lignin-Degrading Fungus Phanerochaete sordida YK-624
White-rot fungi are the most important group of lignin biodegraders. Phanerochaete sordida YK-624 has higher ligninolytic activity than that of model white-rot fungi. However, the underlying mechanism responsible for lignin degradation by white-rot fungi remains unknown, and the induced compounds isolated from white-rot fungi for lignin degradation have never been studied. In the present study, we tried to screen ligninolytic-inducing compounds produced by P. sordida YK-624. After large-scale incubation of P. sordida YK-624, the culture and mycelium were separated by filtration. After the separation and purification, purified compounds were analyzed by high-resolution electrospray ionization mass spectrometry and nuclear magnetic resonance. The sterilized unbleached hardwood kraft pulp was used for the initial evaluation of ligninolytic activity. Ergosterol was isolated and identified and it induced the lignin-degrading activity of this fungus. Moreover, we investigated ergosterol metabolites from P. sordida YK-624, and the ergosterol metabolites ergosta-4,7,22-triene-3,6-dione and ergosta-4,6,8(14),22-tetraen-3-one were identified and then chemically synthesized. These compounds significantly improved the lignin-degrading activity of the fungus. This is the first report on the ligninolytic-inducing compounds produced by white-rot fungi.
Axl and immune checkpoints inhibitors from fruiting bodies of Pleurocybella porrigens
A novel compound (1) and three known ones (2−4) were isolated from the fruiting bodies of Pleurocybella porrigens. The structure of the novel compound was determined by 1D and 2D NMR and HRESIMS data. The biological activity of 1−3 was evaluated using the A549 lung cancer cell line. The results showed the inhibitory activity of compounds 1−3 on the expression of Axl and immune checkpoint molecules.
Bioactive Compounds from the Mushroom-Forming Fungus Chlorophyllum molybdites
A novel compound (1) along with two known compounds (2 and 3) were isolated from the culture broth of Chlorophyllum molybdites, and three known compounds (4–6) were isolated from its fruiting bodies. The planar structure of 1 was determined by the interpretation of spectroscopic data. By comparing the specific rotation of the compound with that of the analog compound, the absolute configuration of 1 was determined to be R. This is the first time that compounds 2–4 were isolated from a mushroom-forming fungus. Compound 2 showed significant inhibition activity against Axl and immune checkpoints (PD-L1, PD-L2). In the bioassay to examine growth inhibitory activity against the phytopathogenic bacteria Peptobacterium carotovorum, Clavibacter michiganensis and Burkholderia glumae, compounds 2 and 3 inhibited the growth of P. carotovorum and C. michiganensis. In the bioassay to examine plant growth regulatory activity, compounds 1–4 showed a significant regulatory activity on lettuce growth.
A New Omics Data Resource of Pleurocybella porrigens for Gene Discovery
Pleurocybellaporrigens is a mushroom-forming fungus, which has been consumed as a traditional food in Japan. In 2004, 55 people were poisoned by eating the mushroom and 17 people among them died of acute encephalopathy. Since then, the Japanese government has been alerting Japanese people to take precautions against eating the P. porrigens mushroom. Unfortunately, despite efforts, the molecular mechanism of the encephalopathy remains elusive. The genome and transcriptome sequence data of P. porrigens and the related species, however, are not stored in the public database. To gain the omics data in P. porrigens, we sequenced genome and transcriptome of its fruiting bodies and mycelia by next generation sequencing. Short read sequences of genomic DNAs and mRNAs in P. porrigens were generated by Illumina Genome Analyzer. Genome short reads were de novo assembled into scaffolds using Velvet. Comparisons of genome signatures among Agaricales showed that P. porrigens has a unique genome signature. Transcriptome sequences were assembled into contigs (unigenes). Biological functions of unigenes were predicted by Gene Ontology and KEGG pathway analyses. The majority of unigenes would be novel genes without significant counterparts in the public omics databases. Functional analyses of unigenes present the existence of numerous novel genes in the basidiomycetes division. The results mean that the omics information such as genome, transcriptome and metabolome in basidiomycetes is short in the current databases. The large-scale omics information on P. porrigens, provided from this research, will give a new data resource for gene discovery in basidiomycetes.
Meta-analysis of neonicotinoid insecticides in global surface waters
Neonicotinoids (NEOs) are a class of insecticides that have high insecticidal activity and are extensively used worldwide. However, increasing evidence suggests their long-term residues in the environment and toxic effects on nontarget organisms. NEO residues are frequently detected in water and consequently have created increasing levels of pollution and pose significant risks to humans. Many studies have focused on NEO concentrations in water; however, few studies have focused on global systematic reviews or meta-analyses of NEO concentrations in water. The purpose of this review is to conduct a meta-analysis on the concentration of NEOs in global waters based on published detections from several countries to extend knowledge on the application of NEOs. In the present study, 43 published papers from 10 countries were indexed for a meta-analysis of the global NEO distribution in water. Most of these studies focus on the intensive agricultural area, such as eastern Asia and North America. The order of mean concentrations is identified as imidacloprid (119.542 ± 15.656 ng L −1 ) > nitenpyram (88.076 ± 27.144 ng L −1 ) > thiamethoxam (59.752 ± 9.068 ng L −1 ) > dinotefuran (31.086 ± 9.275 ng L −1 ) > imidaclothiz (24.542 ± 2.906 ng L −1 ) > acetamiprid (23.360 ± 4.015 ng L −1 ) > thiacloprid (11.493 ± 5.095 ng L −1 ). Moreover, the relationships between NEO concentrations and some environmental factors are analyzed. NEO concentrations increase with temperature, oxidation–reduction potential, and the percentage of cultivated crops but decrease with stream discharge, pH, dissolved oxygen, and precipitation. NEO concentrations show no significant relations to turbidity and conductivity.
Expression of a manganese peroxidase isozyme 2 transgene in the ethanologenic white rot fungus Phlebia sp. strain MG-60
Background The white-rot fungus Phlebia sp. strain MG-60 was proposed as a candidate for integrated fungal fermentation process (IFFP), which unifies aerobic delignification and semi-aerobic consolidated biological processing by a single microorganism based on its ability to efficiently degrade lignin and ferment the sugars from cellulose. To improve IFFP, the development of a molecular breeding method for strain MG-60 is necessary. The purpose of this study is to establish the transformation method for the strain MG-60 and to obtain the over-expressing transformants of lignin-degrading enzyme, manganese peroxidase. Findings In the present study, the expression vector regulated by Phlebia brevispora glyceraldehyde-3-phosphate dehydrogenase promoter and terminator was constructed. A polyethylene glycol transformation method for the ethanol-fermenting white-rot fungus Phlebia sp. MG-60 was established with high transformation efficiency, and the manganese peroxidase isozyme 2 gene (MG mnp2 ) transformants were obtained, showing higher MnP activity than control transformants. MG mnp2 transformants showed higher selective lignin degradation on Quercus wood powder. Conclusions This first report of MG-60 transformation provides a useful methodology for widely accessible to interested researches. These results indicate the possibility of metabolic engineering of strain MG-60 for improving IFFP.
Transformation by complementation of a uracil auxotroph of the hyper lignin-degrading basidiomycete Phanerochaete sordida YK-624
Phanerochaete sordida YK-624 is a hyper lignin-degrading basidiomycete possessing greater ligninolytic selectivity than either P. chrysosporium or Trametes versicolor. To construct a gene transformation system for P. sordida YK-624, uracil auxotrophic mutants were generated using a combination of ultraviolet (UV) radiation and 5-fluoroorotate resistance as a selection scheme. An uracil auxotrophic strain (UV-64) was transformed into a uracil prototroph using the marker plasmid pPsURA5 containing the orotate phosphoribosyltransferase gene from P. sordida YK-624. This system generated approximately 50 stable transformants using 2 x 10⁷ protoplasts. Southern blot analysis demonstrated that the transformed pPsURA5 was ectopically integrated into the chromosomal DNA of all transformants. The enhanced green fluorescent protein (EGFP) gene was also introduced into UV-64. The transformed EGFP was expressed in the co-transformants driven by P. sordida glyceraldehyde-3-phosphate dehydrogenase gene promoter and terminator regions.