Ependymoma is a rare and chemotherapy-resistant brain tumor, which has resulted in a delay in the development of drugs to treat it. A subclass of supratentorial ependymomas (ST-EPN), designated ST-EPN-zinc finger-tran...Ependymoma is a rare and chemotherapy-resistant brain tumor, which has resulted in a delay in the development of drugs to treat it. A subclass of supratentorial ependymomas (ST-EPN), designated ST-EPN-zinc finger-translocation-associated (ZFTA, ST-EPN-ZFTA), exhibits the expression of a fusion protein comprising ZFTA and v-rel reticuloendotheliosis viral oncogene homolog A (RELA), an effector transcription factor of the nuclear factor-kappa B (NF-κB) pathway (ZFTA-RELA). The expression of ZFTA-RELA results in the hyperactivation of the oncogenic NF-κB signaling pathway, which ultimately leads to the development of ST-EPN-ZFTA. To identify inhibitors of the NF-κB signaling pathway activated by the expression of ZFTA-RELA, we used a doxycycline-inducible ZFTA-RELA-expressing NF-κB reporter cell line and found that extracts of the fungus Neosartorya spinosa IFM 47025 exhibited NF-κB inhibitory activity. We identified eight compounds [aszonapyrone A (2), sartorypyrone A (3), epiheveadride (4), acetylaszonalenin (5), (R)-benzodiazepinedione (6), aszonalenin (7), sartorypyrone E (8) and (Z, Z)-N,N’-(1,2-bis[(4-methoxyphenyl)methylene]-1,2-ethanediyl)bis-formamide (9)] from N. spinosa IFM 47025 culture extract using a variety of chromatographic techniques. The structures of these compounds were identified through the analysis of various instrumental data (1D, 2D-NMR, MS, and optical rotation). The NF-κB responsive reporter assay indicated that compounds 2, 3, 5, 7, and 9 exhibited inhibitory activity. We further evaluated the inhibitory activity of these compounds against the expression of endogenous NF-κB responsive genes (CCND1, L1CAM, ICAM1, and TNF) and found that compound 2 showed significant inhibitory activity. Further studies are required to elucidate the mechanism of action of compound 2, which may serve as a lead compound for the development of a novel therapy for ST-EPN-ZFTA.展开更多
Small, cysteine-rich, highly stable antifungal proteins secreted by filamentous Ascomycetes have great po- tential for the development of novel antifungal strate- gies. However, their practical application is still li...Small, cysteine-rich, highly stable antifungal proteins secreted by filamentous Ascomycetes have great po- tential for the development of novel antifungal strate- gies. However, their practical application is still limited due to their not fully clarified mode of action. The aim of this work was to provide a deep insight into the anti-fungal mechanism of Neosartorya fischeri antifungal protein （NFAP）, a novel representative of this protein group. Within a short exposure time to NFAP, reduced cellular metabolism, apoptosis induction, changes in the actin distribution and chitin deposition at the hyphal tip were observed in NFAP-sensitive Aspergillus nidulans. NFAP did show neither a direct membrane disrupting- effect nor uptake by endocytosis. Investigation of A. nidulans signalling mutants revealed that NFAP acti- vates the cAMP/protein kinase A pathway via G-protein signalling which leads to apoptosis and inhibition of polar growth. In contrast, NFAP does not have any in- fluence on the cell wall integrity pathway, but an un- known cell wall integrity pathway-independent mitogen activated protein kinase A-activated target is assumed to be involved in the cell death induction. Taken to- gether, it was concluded that NFAP shows similarities, but also differences in its mode of antifungal action compared to two most investigated NFAP-related pro-teins from Aspergillus giganteus and Penicillium chrysogenum.展开更多
Fungal genomes carry many gene clusters seemingly capable of natural products biosynthesis,yet most clusters remain cryptic or down-regulated. Genome mining revealed an unconventional paraherquonin-like meroterpenoid ...Fungal genomes carry many gene clusters seemingly capable of natural products biosynthesis,yet most clusters remain cryptic or down-regulated. Genome mining revealed an unconventional paraherquonin-like meroterpenoid biosynthetic gene cluster in the chromosome of Neosartorya glabra.The cryptic or down-regulated pathway was activated by constitutive expression of pathway-specific regulator gene ber A encoded within ber biosynthetic gene cluster. Chemical analysis of mutant Ng-OE:ber A extracts enabled the isolation of four berkeleyacetal congeners, in which two of them are new. On the basis of careful bioinformatic analysis of the coding enzymes in the ber gene cluster, the biosynthetic pathway of berkeleyacetals was proposed. These results indicate that this approach would be valuable for discovery of novel natural products and will accelerate the exploitation of prodigious natural products in filamentous fungi.展开更多
A mutualistic relationship between grasses, coal-degrading fungi, and arbuscular mycorrhizal fungi was pro- posed to account for the phyto-biodegradation of coal discard. In this study pot trial experiments were carri...A mutualistic relationship between grasses, coal-degrading fungi, and arbuscular mycorrhizal fungi was pro- posed to account for the phyto-biodegradation of coal discard. In this study pot trial experiments were carried out to confirm transformation of the carbonaceous substrate, in the presence of a suite of coal degrading fungi and arbuscular mycorrhizal fungi, into a humic-enriched soil-like material in the Cynodon dactylon/coal rhizosphere. The results show that after 47 weeks of C. dactylon growth on coal discard the concentration of humics increased from （62.9 -4- 1.5） to （112.1 ＋ 5.4） mg/kg. Substrate humic acid-like substance concentration positively correlated （r2 = 0.95） with accumu- lation of above ground C. dactylon biomass. FTIR spectroscopy of the extracted humic-like substances confirmed both product identity and increased oxidation of the coal discard substrate. Substrate ash content and electrical conductivity declined coincident with an increase in humic acid-like substance concentration, which together reduced the intensity of acidity in the C. dactylon/coal discard rhizosphere. These observations support the proposal that biological oxidative degradation of coal discard leads to increased humic-like substance concentration and formation of a soil-like material. Results have profound implications for use of coal discard as an organic substrate to replace topsoil in phyto-bioreme- diation strategies for sustainable large-scale rehabilitation of coal discard dumps.展开更多
The Aspergillus viridinutans complex includes morphologically similar,soil-inhabiting species.Although its species boundaries have not been fully defined,many isolates from the complex have been isolated as opportunis...The Aspergillus viridinutans complex includes morphologically similar,soil-inhabiting species.Although its species boundaries have not been fully defined,many isolates from the complex have been isolated as opportunistic human and animal pathogens.In the present study,these species were dominant in spoil sites subjected to various types of reclamation management after coal mining.These species were characterised using two different PCR-fingerprinting methods,sequence data from the β-tubulin(benA)and calmodulin(caM)genes,macro-and micromorphology(optical and scanning electron microscopy),maximum growth temperatures and mating experiments.In addition,RNA polymerase II gene(RPB2),actin(act1)and ITS sequences were deposited for the ex-type isolates of newly described species.The mating experiment results,phylogenetic analyses and ascospore morphology suggested the presence of five species in the A.viridinutans complex.Aspergillus aureolus(syn.Neosartorya aureola)was the only homothallic species.Three species,A.felis,A.udagawae(syn.N.udagawae)and A.wyomingensis sp.nov.,were heterothallic and their morphologically distinguishable teleomorph was induced by systematic mating experiments.Aspergillus viridinutans s.str.seems to be a very rare species and was represented only by the ex-type isolate in which the MAT1-1 locus was amplified.Aspegillus viridinutans and A.aureolus were typified in accordance with the rules of the new botanical code.Other species outside the A.viridinutans complex isolated from the reclamation sites were A.fumigatiaffinis and A.lentulus as well as two new sister species,A.brevistipitatus sp.nov.and A.conversis sp.nov.which were closely related each to other and to N.papuensis.Both new species are phylogenetically distant from all anamorphic species and resemble A.brevipes,A.duricaulis and A.unilateralis in micromorphology and are distinguishable from each other by the slower growth of A.conversis on all tested media.Interestingly,no isolate from the reclamation sites represented A.fumigatus s.str.which is usually reported as the dominant species from the section Fumigati in soil.展开更多
文摘为获得酶学性质优良的α-半乳糖苷酶,研究根据Gen Bank数据库提供的嗜热微生物Neosartorya fischeri的α-半乳糖苷酶的氨基酸序列,经密码子优化合成该酶的编码基因。将合成的基因转化大肠杆菌BL21进行原核表达,获得重组α-半乳糖苷酶,并对酶学性质进行研究。结果显示:以棉籽糖为底物时,酶的最适反应温度为55℃,最适p H 6.0,有较好的热稳定性和酸碱稳定性。Fe2+(铁)、Mn2+(锰)和Ca2+(钙)离子可使酶活性大幅提高,而Cu2+(铜)离子则可强烈抑制酶的活性,Pb2+(铅)、Ba2+(钡)、Al3+(铝)、EDTA(乙二胺四乙酸)和SDS(十二烷基硫酸钠)对酶活也有一定的抑制作用。在模拟胃肠环境时发现该酶有良好的稳定性,肠液对该酶甚至有轻微的激活作用。
文摘Ependymoma is a rare and chemotherapy-resistant brain tumor, which has resulted in a delay in the development of drugs to treat it. A subclass of supratentorial ependymomas (ST-EPN), designated ST-EPN-zinc finger-translocation-associated (ZFTA, ST-EPN-ZFTA), exhibits the expression of a fusion protein comprising ZFTA and v-rel reticuloendotheliosis viral oncogene homolog A (RELA), an effector transcription factor of the nuclear factor-kappa B (NF-κB) pathway (ZFTA-RELA). The expression of ZFTA-RELA results in the hyperactivation of the oncogenic NF-κB signaling pathway, which ultimately leads to the development of ST-EPN-ZFTA. To identify inhibitors of the NF-κB signaling pathway activated by the expression of ZFTA-RELA, we used a doxycycline-inducible ZFTA-RELA-expressing NF-κB reporter cell line and found that extracts of the fungus Neosartorya spinosa IFM 47025 exhibited NF-κB inhibitory activity. We identified eight compounds [aszonapyrone A (2), sartorypyrone A (3), epiheveadride (4), acetylaszonalenin (5), (R)-benzodiazepinedione (6), aszonalenin (7), sartorypyrone E (8) and (Z, Z)-N,N’-(1,2-bis[(4-methoxyphenyl)methylene]-1,2-ethanediyl)bis-formamide (9)] from N. spinosa IFM 47025 culture extract using a variety of chromatographic techniques. The structures of these compounds were identified through the analysis of various instrumental data (1D, 2D-NMR, MS, and optical rotation). The NF-κB responsive reporter assay indicated that compounds 2, 3, 5, 7, and 9 exhibited inhibitory activity. We further evaluated the inhibitory activity of these compounds against the expression of endogenous NF-κB responsive genes (CCND1, L1CAM, ICAM1, and TNF) and found that compound 2 showed significant inhibitory activity. Further studies are required to elucidate the mechanism of action of compound 2, which may serve as a lead compound for the development of a novel therapy for ST-EPN-ZFTA.
文摘聚半乳糖醛酸酶在食品和饲料行业具有很大的应用价值,发现新的聚半乳糖醛酸酶不仅能够补充该酶的相关信息,还能为工业应用提供更多选择。从一株嗜热费希新萨托菌（Neosartorya fischeri P1）的基因组中克隆得到一个外切聚半乳糖醛酸酶基因（Nf-pg）,将Nf-pg在毕赤酵母GS115中进行异源表达,重组酶Nf-PG纯化后进行性质测定。Nf-PG在p H 4.5和55℃时活性最高,在p H 2.0~9.0具有较好的稳定性,50℃热处理60 min仍能保持80%的剩余酶活;去除N-糖基化的酶蛋白Nf-PG-D最适温度为50℃,热稳定性降低,最适p H和酸碱稳定性无明显改变。产物分析证明Nf-PG是严格的外切聚半乳糖醛酸酶,优异的性质使其成为食品加工行业的良好选择。
文摘Small, cysteine-rich, highly stable antifungal proteins secreted by filamentous Ascomycetes have great po- tential for the development of novel antifungal strate- gies. However, their practical application is still limited due to their not fully clarified mode of action. The aim of this work was to provide a deep insight into the anti-fungal mechanism of Neosartorya fischeri antifungal protein （NFAP）, a novel representative of this protein group. Within a short exposure time to NFAP, reduced cellular metabolism, apoptosis induction, changes in the actin distribution and chitin deposition at the hyphal tip were observed in NFAP-sensitive Aspergillus nidulans. NFAP did show neither a direct membrane disrupting- effect nor uptake by endocytosis. Investigation of A. nidulans signalling mutants revealed that NFAP acti- vates the cAMP/protein kinase A pathway via G-protein signalling which leads to apoptosis and inhibition of polar growth. In contrast, NFAP does not have any in- fluence on the cell wall integrity pathway, but an un- known cell wall integrity pathway-independent mitogen activated protein kinase A-activated target is assumed to be involved in the cell death induction. Taken to- gether, it was concluded that NFAP shows similarities, but also differences in its mode of antifungal action compared to two most investigated NFAP-related pro-teins from Aspergillus giganteus and Penicillium chrysogenum.
基金supported financially by the National Natural Science Foundation of China (No. 81522043)CAMS Initiative for Innovative Medicine (2017-I2M-4-004)the Thousand Young Talents Program of China
文摘Fungal genomes carry many gene clusters seemingly capable of natural products biosynthesis,yet most clusters remain cryptic or down-regulated. Genome mining revealed an unconventional paraherquonin-like meroterpenoid biosynthetic gene cluster in the chromosome of Neosartorya glabra.The cryptic or down-regulated pathway was activated by constitutive expression of pathway-specific regulator gene ber A encoded within ber biosynthetic gene cluster. Chemical analysis of mutant Ng-OE:ber A extracts enabled the isolation of four berkeleyacetal congeners, in which two of them are new. On the basis of careful bioinformatic analysis of the coding enzymes in the ber gene cluster, the biosynthetic pathway of berkeleyacetals was proposed. These results indicate that this approach would be valuable for discovery of novel natural products and will accelerate the exploitation of prodigious natural products in filamentous fungi.
文摘A mutualistic relationship between grasses, coal-degrading fungi, and arbuscular mycorrhizal fungi was pro- posed to account for the phyto-biodegradation of coal discard. In this study pot trial experiments were carried out to confirm transformation of the carbonaceous substrate, in the presence of a suite of coal degrading fungi and arbuscular mycorrhizal fungi, into a humic-enriched soil-like material in the Cynodon dactylon/coal rhizosphere. The results show that after 47 weeks of C. dactylon growth on coal discard the concentration of humics increased from （62.9 -4- 1.5） to （112.1 ＋ 5.4） mg/kg. Substrate humic acid-like substance concentration positively correlated （r2 = 0.95） with accumu- lation of above ground C. dactylon biomass. FTIR spectroscopy of the extracted humic-like substances confirmed both product identity and increased oxidation of the coal discard substrate. Substrate ash content and electrical conductivity declined coincident with an increase in humic acid-like substance concentration, which together reduced the intensity of acidity in the C. dactylon/coal discard rhizosphere. These observations support the proposal that biological oxidative degradation of coal discard leads to increased humic-like substance concentration and formation of a soil-like material. Results have profound implications for use of coal discard as an organic substrate to replace topsoil in phyto-bioreme- diation strategies for sustainable large-scale rehabilitation of coal discard dumps.
基金This study was supported by the project ME 08085 KONTAKT Soil organisms in spoils after brown coal mining in the U.S.A.and Europe:a bioindication potential and a role in pedogenesisThis research was also partially supported by the Ministry of Education,Youth and Sports(CZ.1.07/2.3.00/20.0055 and CZ.1.07/2.3.00/30.0003)Molecular genetics analyses were supported by the project GAUK 607812.
文摘The Aspergillus viridinutans complex includes morphologically similar,soil-inhabiting species.Although its species boundaries have not been fully defined,many isolates from the complex have been isolated as opportunistic human and animal pathogens.In the present study,these species were dominant in spoil sites subjected to various types of reclamation management after coal mining.These species were characterised using two different PCR-fingerprinting methods,sequence data from the β-tubulin(benA)and calmodulin(caM)genes,macro-and micromorphology(optical and scanning electron microscopy),maximum growth temperatures and mating experiments.In addition,RNA polymerase II gene(RPB2),actin(act1)and ITS sequences were deposited for the ex-type isolates of newly described species.The mating experiment results,phylogenetic analyses and ascospore morphology suggested the presence of five species in the A.viridinutans complex.Aspergillus aureolus(syn.Neosartorya aureola)was the only homothallic species.Three species,A.felis,A.udagawae(syn.N.udagawae)and A.wyomingensis sp.nov.,were heterothallic and their morphologically distinguishable teleomorph was induced by systematic mating experiments.Aspergillus viridinutans s.str.seems to be a very rare species and was represented only by the ex-type isolate in which the MAT1-1 locus was amplified.Aspegillus viridinutans and A.aureolus were typified in accordance with the rules of the new botanical code.Other species outside the A.viridinutans complex isolated from the reclamation sites were A.fumigatiaffinis and A.lentulus as well as two new sister species,A.brevistipitatus sp.nov.and A.conversis sp.nov.which were closely related each to other and to N.papuensis.Both new species are phylogenetically distant from all anamorphic species and resemble A.brevipes,A.duricaulis and A.unilateralis in micromorphology and are distinguishable from each other by the slower growth of A.conversis on all tested media.Interestingly,no isolate from the reclamation sites represented A.fumigatus s.str.which is usually reported as the dominant species from the section Fumigati in soil.
