Publications

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genome mining of a prenylated and immunosuppressive polyketide from pathogenic fungi.activation of the polycyclic polyketide prenyltransferase (pcptase)-containing silent clusters in aspergillus fumigatus and neosartorya fischeri led to isolation of a new metabolite neosartoricin (3). the structure of 3 was solved by x-ray crystallography and nmr to be a prenylated anthracenone. 3 exhibits t-cell antiproliferative activity with an ic(50) of 3 μm, suggestive of a physiological role as an immunosuppressive agent.201323368997
complexity generation in fungal peptidyl alkaloid biosynthesis: a two-enzyme pathway to the hexacyclic mdr export pump inhibitor ardeemin.ardeemins are hexacyclic peptidyl alkaloids isolated from aspergillus fischeri as agents that block efflux of anticancer drugs by multidrug resistance (mdr) export pumps. to evaluate the biosynthetic logic and enzymatic machinery for ardeemin framework assembly, we sequenced the a. fischeri genome and identified the ardabc gene cluster. through both genetic deletions and biochemical characterizations of purified arda and ardb we show this ardab enzyme pair is sufficient to convert anthranilate ( ...201323330675
disparate proteome responses of pathogenic and nonpathogenic aspergilli to human serum measured by activity-based protein profiling (abpp).aspergillus fumigatus is the primary pathogen causing the devastating pulmonary disease invasive aspergillosis in immunocompromised individuals. there is high genomic synteny between a. fumigatus and closely related rarely pathogenic neosartorya fischeri and aspergillus clavatus genomes. we applied activity-based protein profiling to compare unique or overexpressed activity-based probe-reactive proteins of all three fungi over time in minimal media growth and in response to human serum. we found ...201323599423
functionality and prevalence of trehalose-based oligosaccharides as novel compatible solutes in ascospores of neosartorya fischeri (aspergillus fischeri) and other fungi.ascospores of neosartorya, byssochlamys and talaromyces can be regarded as the most stress-resistant eukaryotic cells. they can survive exposure at temperatures as high as 85°c for 100 min or more. neosartorya fischeri ascospores are more viscous and more resistant to the combined stress of heat and desiccation than the ascospores of talaromyces macrosporus which contain predominantly trehalose. these ascospores contain trehalose-based oligosaccharides (tos) that are novel compatible solutes, wh ...201425040129
occurrence of azole-resistant species of aspergillus in the uk environment.the aim of this study was to survey environmental isolates of aspergillus resistant to azoles in azole-treated and naïve areas to determine whether resistance could be related to azole treatment history. aspergillus fumigatus was sampled from the centre of a large city and from fields with known azole history. azole resistance was determined and sequencing was performed to identify strains and mutations in the cyp51a gene. azole resistance was detected in azole-treated field isolates but not in ...201427873687
fungal colonization and enzyme-mediated metabolism of waste coal by neosartorya fischeri strain eccn 84.colonization and oxidative metabolism of south african low-rank discard coal by the fungal strain eccn 84 previously isolated from a coal environment and identified as neosartorya fischeri was investigated. results show that waste coal supported fungal growth. colonization of waste coal particles by n. fischeri eccn 84 was associated with the formation of compact spherical pellets or sclerotia-like structures. dissection of the pellets from liquid cultures revealed a nucleus of "engulfed" coal w ...201425273148
molecular characterization of a highly-active thermophilic β-glucosidase from neosartorya fischeri p1 and its application in the hydrolysis of soybean isoflavone glycosides.isoflavone occurs abundantly in leguminous seeds in the form of glycoside and aglycone. however, isoflavone glycoside has anti-nutritional effect and only the free type is beneficial to human health. in the present study we identified a β-glucosidase from thermophilic neosartorya fischeri p1, termed nfbgl1, capable of efficiently converting isoflavone glycosides into free isoflavones. the gene, belonging to glycoside hydrolase family 3, was successfully overexpressed in pichia pastoris at high c ...201425188254
survey of molds, yeast and alicyclobacillus spp. from a concentrated apple juice productive process.bacteria and molds may spoil and/or contaminate apple juice either by direct microbial action or indirectly by the uptake of metabolites as off-flavours and toxins. some of these microorganisms and/or metabolites may remain in the food even after extensive procedures. this study aim to identify the presence of molds (including heat resistant species) and alicyclobacillus spp., during concentrated apple juice processing. molds were isolated at different steps and then identified by their macrosco ...201424948913
a promiscuous prenyltransferase from aspergillus oryzae catalyses c-prenylations of hydroxynaphthalenes in the presence of different prenyl donors.prenyltransferases of the dimethylallyltryptophan synthase (dmats) superfamily are involved in the biosynthesis of secondary metabolites and show broad substrate specificity towards their aromatic substrates with a high regioselectivity for the prenylation reactions. most members of this superfamily accepted as prenyl donor exclusively dimethylallyl diphosphate (dmapp). one enzyme, anapt from neosartorya fischeri, was reported recently to use both dmapp and geranyl diphosphate (gpp) as prenyl do ...201424430210
mannitol is essential for the development of stress-resistant ascospores in neosartorya fischeri (aspergillus fischeri).the polyol mannitol is one of the main compatible solutes in neosartorya fischeri and accumulates in conidia and ascospores. here, it is shown that biosynthesis of mannitol in n. fischeri mainly depends on mannitol 1-phosphate dehydrogenase (mpda). reporter studies and qpcr analysis demonstrated that mpda is moderately expressed in vegetative hyphae and conidiophores, while it is highly expressed during development of ascospores. deletion of mpda reduced mannitol in whole cultures as much as 85% ...201424412483
a thermophilic α-galactosidase from neosartorya fischeri p1 with high specific activity, broad substrate specificity and significant hydrolysis ability of soymilk.an extracellular α-galactosidase (gal27a) with high specific activity of 423umg(-1) was identified in thermophilic neosartorya fischeri p1. its coding gene (1680bp) was cloned and functionally expressed in pichia pastoris. sequence analysis indicated that deduced gal27a contains a catalytic domain of glycoside hydrolase family 27. the native and recombinant enzymes shared some similar properties, such as ph optima at 4.5, temperature optima at 60-70°c, resistance to most chemicals and saccharide ...201424360500
genome mining for new α-amylase and glucoamylase encoding sequences and high level expression of a glucoamylase from talaromyces stipitatus for potential raw starch hydrolysis.mining fungal genomes for glucoamylase and α-amylase encoding sequences led to the selection of 23 candidates, two of which (designated tsgam-2 and nfamy-2) were advanced to testing for cooked or raw starch hydrolysis. tsgam-2 is a 66-kda glucoamylase recombinantly produced in pichia pastoris and originally derived for talaromyces stipitatus. when harvested in a 20-l bioreactor at high cell density (od600 > 200), the secreted tsgam-2 enzyme activity from p. pastoris strain gs115 reached 800 u/ml ...201424046254
production of a defensin-like antifungal protein nfap from neosartorya fischeri in pichia pastoris and its antifungal activity against filamentous fungal isolates from human infections.neosartorya fischeri nrrl 181 isolate secretes a defensin-like antifungal protein (nfap) which has a remarkable antifungal effect against ascomycetous filamentous fungi. this protein is a promising antifungal agent of biotechnological value; however in spite of the available knowledge of the nature of its 5'-upstream transcriptional regulation elements, the bulk production of nfap has not been resolved yet. in this study we carried out its heterologous expression in the yeast pichia pastoris and ...201424269762
a decrease in bulk water and mannitol and accumulation of trehalose and trehalose-based oligosaccharides define a two-stage maturation process towards extreme stress resistance in ascospores of neosartorya fischeri (aspergillus fischeri).fungal propagules survive stresses better than vegetative cells. neosartorya fischeri, an aspergillus teleomorph, forms ascospores that survive high temperatures or drying followed by heat. not much is known about maturation and development of extreme stress resistance in fungal cells. this study provides a novel two-step model for the acquisition of extreme stress resistance and entry into dormancy. ascospores of 11- and 15-day-old cultures exhibited heat resistance, physiological activity, acc ...201525040022
microarray analysis of neosartorya fischeri using different carbon sources, petroleum asphaltenes and glucose-peptone.asphaltenes are considered as the most recalcitrant petroleum fraction and represent a big problem for the recovery, separation and processing of heavy oils and bitumens. neosartorya fischeri is a saprophytic fungus that is able to grow using asphaltenes as the sole carbon source [1]. we performed transcription profiling using a custom designed microarray with the complete genome from n. fischeri nrrl 181 in order to identify genes related to the transformation of asphaltenes [1]. data analysis ...201526484261
a novel gh7 endo-β-1,4-glucanase from neosartorya fischeri p1 with good thermostability, broad substrate specificity and potential application in the brewing industry.an endo-β-1,4-glucanase gene, cel7a, was cloned from the thermophilic cellulase-producing fungus neosartorya fischeri p1 and expressed in pichia pastoris. the 1,410-bp full-length gene encodes a polypeptide of 469 amino acids consisting of a putative signal peptide at residues 1-20, a catalytic domain of glycoside hydrolase family 7 (gh7), a short thr/ser-rich linker and a family 1 carbohydrate-binding module (cbm 1). the purified recombinant cel7a had ph and temperature optima of ph 5.0 and 60° ...201526360701
genome mining for sesterterpenes using bifunctional terpene synthases reveals a unified intermediate of di/sesterterpenes.genome mining is a promising method to discover novel secondary metabolites in the postgenomic era. we applied the aspergillus oryzae heterologous expression system to functionally characterize cryptic bifunctional terpene synthase genes found in fungal genomes and identified the sesterfisherol synthase gene (nfss) from neosartorya fischeri. sesterfisherol contains a characteristic 5-6-8-5 tetracyclic ring system and is modified by cytochrome p450 monooxygenase (nfp450) to sesterfisheric acid. t ...201526332841
biochemical characterization of three distinct polygalacturonases from neosartorya fischeri p1.polygalacturonase is one of the most important industrial pectinases. to enrich the genetic resources and develop new enzyme candidates, three polygalacturonase genes (nfpg i-iii) of glycosyl hydrolase family 28 were cloned from neosartorya fischeri p1 and functionally expressed in pichia pastoris. the purified recombinant proteins exhibited some distinguished properties. in comparison with other counterparts, nfpg i showed the highest specific activity (40, 123 u/mg), nfpg ii had the highest te ...201526041232
insight into the antifungal mechanism of neosartorya fischeri antifungal protein.small, cysteine-rich, highly stable antifungal proteins secreted by filamentous ascomycetes have great potential for the development of novel antifungal strategies. 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 antifungal mechanism of neosartorya fischeri antifungal protein (nfap), a novel representative of this protein group. within a short exposure time to nfap, reduced cellular ...201525994413
structural analysis of novel trehalose-based oligosaccharides from extremely stress-tolerant ascospores of neosartorya fischeri (aspergillus fischeri).different fungi, including the genera neosartorya, byssochlamys and talaromyces, produce (asco)spores that survive pasteurization treatments and are regarded as the most stress-resistant eukaryotic cells. here, the nmr analysis of a series of trehalose-based oligosaccharides, being compatible solutes that are accumulated to high levels in ascospores of the fungus neosartorya fischeri, is presented. these oligosaccharides consist of an α,α-trehalose backbone, extended with one [α-d-glcp-(1 → 6)-α ...201525974853
the first case of feline sinonasal aspergillosis due to aspergillus fischeri in japan.feline upper respiratory tract infection due to aspergillus spp. is considered an emerging disease, with the number of reported cases continuing to rise. in this study, we report the first case of feline sinonasal aspergillosis caused by aspergillus fischeri in japan. the patient presented after 2 months of progressive facial deformity around the nose and nasal discharge. the isolate from this case was susceptible to itraconazole (itz), voriconazole and micafungin, but was resistant to amphoteri ...201525947561
targeted production of secondary metabolites by coexpression of non-ribosomal peptide synthetase and prenyltransferase genes in aspergillus.prenylated tryptophan-containing cyclic dipeptides are found in different fungi and serve as precursors for the biosynthesis of diverse biologically active secondary metabolites. they show distinct and usually higher biological and pharmacological activities than the respective non-prenylated dipeptides. successful production of such compounds were achieved by a new approach based on the coexpression of ftmps, a non-ribosomal peptide synthetase from neosartorya fischeri, with three cyclic dipept ...201525744649
sartorypyrone d: a new nadh-fumarate reductase inhibitor produced by neosartorya fischeri fo-5897. 201525586023
erratum to: insights into the substrate specificity and synergy with mannanase of family 27 α-galactosidases from neosartorya fischeri p1. 201525573475
avenaciolides: potential mura-targeted inhibitors against peptidoglycan biosynthesis in methicillin-resistant staphylococcus aureus (mrsa).discovery of new antibiotics for combating methicillin-resistant staphylococcus aureus (mrsa) is of vital importance in the post-antibiotic era. here, we report four avenaciolide derivatives (1-4) isolated from neosartorya fischeri, three of which had significant antimicrobial activity against mrsa. the morphology of avenaciolide-treated cells was protoplast-like, which indicated that cell wall biosynthesis was interrupted. comparing the structures and minimum inhibitory concentrations of 1-4, t ...201525521652
insights into the substrate specificity and synergy with mannanase of family 27 α-galactosidases from neosartorya fischeri p1.thermophilic neosartorya fischeri p1 is an excellent carbohydrate-active enzyme (cazyme) producer. two α-galactosidases of gh (glycoside hydrolase ) family 27 with a very low sequence identity (28.7%), gal27a and gal27b, were identified in strain p1 and functionally expressed in pichia pastoris. in comparison to other characterized gh27 fungal counterparts, rgal27b has a higher temperature optimum (75 °c) and better thermostability (>50% activity at 70 °c for 15 min), and rgal27a shows stability ...201525487894
a thermophilic β-mannanase from neosartorya fischeri p1 with broad ph stability and significant hydrolysis ability of various mannan polymers.a new β-mannanase gene, man5p1, was cloned from the thermophilic fungus neosartorya fischeri p1, and successfully expressed in pichia pastoris. the predicted amino acid sequence of man5p1 consists of a putative 19-residue signal peptide at the n-terminus and a catalytic domain of glycoside hydrolase family 5. the purified recombinant man5p1 (rman5p1) was optimally active at ph 4.0 and 80 °c, and was acid and alkali tolerant, exhibiting >20% of the maximal activity at ph 2.0 and 9.0. rman5p1 had ...201525466024
modeling the growth of byssochlamys fulva and neosartorya fischeri on solidified apple juice by measuring colony diameter and ergosterol content.byssochlamys fulva and neosartorya fischeri are heat-resistant fungi which are a concern to food industries (e.g. apple juice industry) since their growth represents significant economic liabilities. although the most common method used to assess fungal growth in solid substrates is by measuring the colony's diameter, it is difficult to apply this method to food substrates. alternatively, ergosterol contents have been used to quantify fungal contamination in some types of food. the current study ...201525462919
enhanced biological straw saccharification through coculturing of lignocellulose-degrading microorganisms.lignocellulosic waste (lcw) is an abundant, low-cost, and inedible substrate for the induction of lignocellulolytic enzymes for cellulosic bioethanol production using an efficient, environmentally friendly, and economical biological approach. in this study, 30 different lignocellulose-degrading bacterial and 18 fungal isolates were quantitatively screened individually for the saccharification of four different ball-milled straw substrates: wheat, rice, sugarcane, and pea straw. rice and sugarcan ...201525724976
revisiting overexpression of a heterologous β-glucosidase in trichoderma reesei: fusion expression of the neosartorya fischeri bgl3a to cbh1 enhances the overall as well as individual cellulase activities.the filamentous fungus trichoderma reesei has the capacity to secret large amounts of cellulase and is widely used in a variety of industries. however, the t. reesei cellulase is weak in β-glucosidase activity, which results in accumulation of cellobiose inhibiting the endo- and exo-cellulases. by expressing an exogenous β-glucosidase gene, the recombinant t. reesei cellulase is expected to degrade cellulose into glucose more efficiently.201627400964
aspergillus fumigatus-related species in clinical practice.aspergillus fumigatus is the main etiologic agent of invasive aspergillosis (ia). other aspergillus species belonging to the section fumigati (a. fumigatus complex) may occasionally be the cause of ia. these strains are often misidentified, as they cannot be distinguished from a. fumigatus by conventional morphological analysis and sequencing methods. this lack of recognition may have important consequences as these a. fumigatus-related species often display some level of intrinsic resistance to ...201627242710
[neosartorya fischeri (wehmer) malloch & cain]. 201626965879
a penicillium chrysogenum-based expression system for the production of small, cysteine-rich antifungal proteins for structural and functional analyses.small, cysteine-rich and cationic antifungal proteins (aps) from filamentous ascomycetes, such as nfap from neosartorya fischeri and paf from penicillium chrysogenum, are promising candidates for novel drug development. a prerequisite for their application is a detailed knowledge about their structure-function relation and mode of action, which would allow protein modelling to enhance their toxicity and specificity. technologies for structure analyses, such as electronic circular dichroism (ecd) ...201627835989
heterologous production of an acidic thermostable lipase with broad-range ph activity from thermophilic fungus neosartorya fischeri p1.thermophilic neosartorya fischeri p1 is an excellent lipase producer and harbors seven lipase genes. all genes were found to be functional after heterologous expression in escherichia coli. one of them, lip09, showed high-level expression in pichia pastoris with the yield of 2.0 g/l in a 3.7-l fermentor. deduced amino acid sequence of lip09 consists of a putative signal peptide (residues 1-19) and a mature polypeptide (residues 20-562). compared with other fungal counterparts, purified recombina ...201627503553
biotransformation of petroleum asphaltenes and high molecular weight polycyclic aromatic hydrocarbons by neosartorya fischeri.neosartorya fischeri, an aspergillaceae fungus, was evaluated in its capacity to transform high molecular weight polycyclic aromatics hydrocarbons (hmw-pahs) and the recalcitrant fraction of petroleum, the asphaltenes. n. fischeri was able to grow in these compounds as sole carbon source. coronene, benzo(g,h,i)perylene, and indeno(1,2,3-c,d)pyrene, together with the asphaltenes, were assayed for fungal biotransformation. the transformation of the asphaltenes and hmw-pahs was confirmed by reverse ...201626893177
comparison of chemical sensitivity of fresh and long-stored heat resistant neosartorya fischeri environmental isolates using biolog phenotype microarray system.spoilage of heat processed food and beverage by heat resistant fungi (hrf) is a major problem for food industry in many countries. neosartorya fischeri is the leading source of spoilage in thermally processed products. its resistance to heat processing and toxigenicity makes studies about neosartorya fischeri metabolism and chemical sensitivity essential. in this study chemical sensitivity of two environmental neosartorya fischeri isolates were compared. one was isolated from canned apples in 19 ...201626815302
hydrophilins in the filamentous fungus neosartorya fischeri (aspergillus fischeri) have protective activity against several types of microbial water stress.hydrophilins are proteins that occur in all domains of life and protect cells and organisms against drought and other stresses. they include most of the late embryogenesis abundant (lea) proteins and the heat shock protein (hsp) hsp12. here, the role of a predicted lea-like protein (leama) and two hsp12 proteins (hsp12a and hsp12b) of neosartorya fischeri was studied. this filamentous fungus forms ascospores that belong to the most stress-resistant eukaryotic cells described to date. heterologou ...201626487515
nfap2, a novel cysteine-rich anti-yeast protein from neosartorya fischeri nrrl 181: isolation and characterization.the increasing incidence of fungal infections and damages due to drug-resistant fungi urges the development of new antifungal strategies. the cysteine-rich antifungal proteins from filamentous ascomycetes provide a feasible base for protection against molds due to their potent antifungal activity on them. in contrast to this, they show no or weak activity on yeasts, hence their applicability against this group of fungi is questionable. in the present study a 5.6 kda anti-yeast protein (nfap2) is ...201627637945
cloning and characterization of f3pyc gene encoding pyruvate carboxylase in aspergillus flavus strain (f3).pyruvate carboxylase is a major enzyme for biosynthesis of organic acids like; citric acid, fumeric acid, and l-malic acid. these organic acids play very important role for biological remediation of heavy metals. in this study, gene walking method was used to clone and characterize pyruvate carboxylase gene (f3pyc) from heavy metal resistant indigenous fungal isolate aspergillus flavus (f3). 3579 bp of an open reading frame which encodes 1193 amino acid protein (isoelectric point: 6.10) with a c ...201728710744
aspergillus fischeri mediated biosynthesis of gold nanoparticles and their beneficially comparative effect on normal and cancer cell lines.background along with the intensified use of metal nanoparticles, growing concern of their adverse outcome on human health has also expanded, indicating that this work is an integral part of nanobioscience study. objective the aim of this study was to evaluate varied effect of biosynthesized gold nanoparticles (aunps) on normal and cancerous mammalian cells. methods aunp synthesized and characterized by different characterization methods, here are produced by specifically isolated aspergillus sp ...201728530537
structural determinants of neosartorya fischeri antifungal protein (nfap) for folding, stability and antifungal activity.the recent global challenges to prevent and treat fungal infections strongly demand for the development of new antifungal strategies. the structurally very similar cysteine-rich antifungal proteins from ascomycetes provide a feasible basis for designing new antifungal molecules. the main structural elements responsible for folding, stability and antifungal activity are not fully understood, although this is an essential prerequisite for rational protein design. in this study, we used the neosart ...201728512317
biochemical properties and evaluation of washing performance in commercial detergent compatibility of two collagenolytic serine peptidases secreted by aspergillus fischeri and penicillium citrinum.filamentous fungi secrete diverse peptidases with different biochemical properties, which is of considerable importance for application in various commercial sectors. in this study, we describe the isolation of two fungal species collected from the soil of decayed organic matter: aspergillus fischeri and penicillium citrinum. in a submerged bioprocess, we observed better peptidase production with the fungus p. citrinum, which reached a peak production at 168 h with 760 u/ml, in comparison with t ...201727552601
crude extracts of marine-derived and soil fungi of the genus neosartorya exhibit selective anticancer activity by inducing cell death in colon, breast and skin cancer cell lines.the crude ethyl acetate extracts of marine-derived fungi neosartorya tsunodae kufc 9213 (e1) and n. laciniosa kufc 7896 (e2), and soil fungus n. fischeri kufc 6344 (e3) were evaluated for their in vitro anticancer activities on a panel of seven human cancer cell lines.201726941530
loop 3 of fungal endoglucanases of glycoside hydrolase family 12 modulates catalytic efficiency.glycoside hydrolase (gh) family 12 comprises enzymes with a wide range of activities critical for the degradation of lignocellulose. however, the important roles of the loop regions of gh12 enzymes in substrate specificity and catalytic efficiency remain poorly understood. this study examined how the loop 3 region affects the enzymatic properties of gh12 glucanases using nfeg12a from neosartorya fischeri p1 and eg (pdb 1ks4) from aspergillus niger acidophilic and thermophilic nfeg12a had the hig ...201728039140
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