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Karakoram mountain range contains the tallest mountain peaks of the world with thousands of glaciers which are microbiologically untapped. This is a pioneer approach of isolation and characterization of fungi from Batura glacier, Karakoram, Pakistan. Total number (CFU/mL or g) was determined at 4 °C and 15 °C to isolate psychrophilic and psychrotrophic fungi, respectively. About 33 different fungi were isolated from sediment (29), ice (2) and water (2) and were identified morphologically and by sequencing of specific internal transcribed spacer region of the species through internal transcribed spacer 1 (ITS1) and internal transcribed spacer 2 (ITS2) primers. Mostly, the fungal isolates belonged to genus Penicillium, followed by Cladosporium, Geomyces, Cordyceps, Mrakia, Cadophora, Tetracladium, Eupenicillium, Trametes, Mortierella, Scopulariopsis, Beauveria, Candida and Pseudogymnoascus. They showed growth at wide temperature range from 4-37 °C, few at 45 °C as well. Most of the isolates were able to grow at pH 1- 13. Fungal isolates were able to grow in 2-26% NaCl, with the highest tolerance (26%) by Mrakia robertii. All study isolates showed inhibitory activity against Staphylococcus sp., and four industrially important enzymes (cellulase, DNase, lipase and phosphatase) were produced by Mrakia robertii. The fungal isolates of such cold habitats are potential sources of various industrial and environmental application. [ABSTRACT FROM AUTHOR]

Three new azaphilones, eupenicilazaphilones A–C ( 1 – 3 ), and one new isocoumarin, eupenicillin A ( 4 ), as well as five known azaphilones were isolated from the sponge-derived fungus Eupenicillium sp. 6A-9. Their structures were elucidated by detailed spectroscopic analysis, comparison with literature data, and 13 C NMR chemical shifts and TDDFT/ECD calculations. The antibacterial activity against Staphylococcus aureus ATCC25923, methicillin-resistant Staphylococcus aureus ATCC4330, and Acinetobacter baumanii ATCC19606 as well as the cytotoxic activity towards human cancer lines MCF-7 and A549 were examined. [ABSTRACT FROM AUTHOR]

We investigated the wood-damaging characteristics of microorganisms by conducting decay tests on the surface of woodblocks made from the wood of Prunus sargentii , with the aim of identifying ways to preserve the woodblocks. The microorganisms were classified into 29 types of fungi and 6 types of bacteria. Among them 11 types of fungi showed high wood-degrading enzyme activity, and hence classified under risk levels 2–3. We also found that the anti-mold effect of P. sargentii was low, based on the finding that surface contaminants such as Eupenicillium rubidurum and Penicillium oxalicum grew on the P. sargentii specimen. In addition, these strains were grew on the surface of a lacquered P. sargentii specimen, and the anti-mold effect of the lacquer did not seem to correlate with the number of lacquer applications. The P. sargentii specimen was found to have a higher weight loss rate than a Pinus densiflora specimen, which indicated that the preservative effect of P. sargentii against Coprinellus radians was low. However, the weight loss rate of the lacquered P. sargentii specimen was low, and mycelium was not observed in the wood tissue. In conclusion, we believe that lacquer exerts a preservative effect on P. sargentii specimens. [ABSTRACT FROM AUTHOR]

The combination of hydrothermal pretreatment (autohydrolysis) and enzymatic hydrolysis was comprehensively evaluated for the efficient release of monosaccharides and ferulic acid from corn bran. Arabinan was depolymerized and solubilized more easily during autohydrolysis compared to xylan, esterified ferulic acid, and the acetyl group. Also, the enzymatic xylose yield showed strong linear correlation with arabinan, ferulic acid, and acetic acid content in autohydrolysis residues while correlations between enzymatic glucose yield and hemicellulose contents were separated into two stages with different slopes. The addition of a few debranching enzymes to commercial cellulase and xylanase only slightly enhanced enzymatic hydrolysis of autohydrolysis residues, whereas an enzyme blend from Aspergillus oryzae and Eupenicillium parvum showed a significant synergistic effect. Desirable combined hydrolysis yields of glucose (72.26%), xylose (75.87%), arabinose (76.95%), and ferulic acid (74.13%) were obtained after autohydrolysis at 165 °C for 40 min and subsequent hydrolysis by an equal mixture blend produced by A. oryzae and E. parvum at an enzyme loading dosage of 14.1 mg protein/g dry destarched corn bran. [ABSTRACT FROM AUTHOR]

The mesophilic fungus Eupenicillium parvum 4–14 is an important producer of thermotolerant hemicellulolytic and cellulolytic enzymes. The aim of this study was to establish a method for genetic manipulation of the fungus by Agrobacterium tumefaciens . The promotor PgpdA of a glyceraldehyde-3-phosphate dehydrogenase gene was isolated from E. parvum 4–14. To transform the fungus, an expression plasmid containing a superfolder green fluorescent protein (sfGFP) gene under the control of PgpdA promotor was constructed using the plasmid pAg1-H3 as a parental plasmid. Using the fungal ascospores as receptor and hygromycin B resistance as a selection marker, the recombinant plasmid was successfully introduced into the fungal cells by A. tumefaciens- mediated transformation (ATMT) method. Acetosyringone (AS) was essential to the successful transformation. The transformation frequency was significantly affected by the co-culture temperature and time, the quantity of fungal spores and the AS concentration. The highest transformation frequency was up to 373 transformants per 10 5 fungal spores, which was higher than those of other fungal species. The fungal transformants were genetically stable after five subcultures in the absence of antibiotic. GFP protein was strongly expressed in the hypha of fungal transformants. In conclusion, the ATMT is a highly efficient method for genetic manipulation of E. parvum 4–14, and will improve the molecular researches on the fungus. [ABSTRACT FROM AUTHOR]

Two rare 6(D)/5(E) ring-fused meroterpenoids, eupeniacetal A (1) and eupeniacetal B (2), and three new meroterpenoids, 1-methoxy-hydropreaustinoid A1 (3), hydroberkeleyone B (4), and 22-deoxy-10-oxominiolutelide B (5), as well as five known meroterpenoids, 22-deoxyminiolutelide B (6), preaustinoid A1 (7), berkeleyone C (8), berkeleyacetal A (9), and preaustinoid A3 (10), have been isolated, through the aid of LC-MS, from the sponge-derived fungus Eupenicillium sp. 6A-9. The structures of these meroterpenoids were elucidated by detailed spectroscopic analysis, comparison with literature data, and single-crystal X-ray diffraction. The absolute configurations were assigned by means of TDDFT/ECD and [a]D calculations as well as by X-ray diffraction analysis. These compounds showed inhibitory activity towards TNF-a production in the induced THP-1 cell line but were inactive against normal tumor cell lines. [ABSTRACT FROM AUTHOR]

The genus Talaromyces constitutes an important group of molds with species that are mainly found in soil, indoor environments and food products. Traditionally, it has been considered, together with Eupenicillium, the teleomorphic state of Penicillium. However, the taxonomy of these fungi has changed considerably, and Talaromyces currently includes sexually and asexually reproducing species. In a previous study of the occurrence of penicillium-like fungi from clinical samples in the USA, we used the combined phylogeny of the internal transcribed spacer ( ITS) region of the rDNA and β-tubulin ( BenA) gene to identify 31 isolates of Talaromyces, 85 of Penicillium and two of Rasamsonia. However, seven isolates of Talaromyces were assigned to the corresponding sections but not to any particular species. In this study, we have resolved the taxonomy of these isolates through a multilocus sequence analysis of the ITS, fragments of the BenA, calmodulin ( CaM), and RNA polymerase II second largest subunit ( RPB2) genes, and a detailed phenotypic study. As a result, four new species are described and illustrated, ie Talaromyces alveolaris, T. georgiensis, T. minnesotensis and T. rapidus. [ABSTRACT FROM AUTHOR]

A putative sucrose transporter PdSUT1 included in the same clade that Sut1p from Schizosaccharomyces pombe was identified in Penicillium digitatum , the major citrus postharvest pathogen. PdSUT1 gene was characterized using target gene disruption and gene overexpression. The Δ PdSUT1 mutants generated by gene elimination showed reduction in fungal virulence during citrus fruit infection assayed in mature fruit at 20 °C. However, the overexpression mutants did not increased disease severity neither in the mutants coming from a high virulent nor from a low virulent P. digitatum progenitor strains. Moreover, fungicide sensitivity was affected in the deletant mutants but not in the overexpression transformants. The expression analysis of several genes involved in fungicide resistance showed an intensification of MFS transporters and a decrease of sterol demethylases transcriptional abundance in the Δ PdSUT1 mutants compare to the parental wild type strain. PdSUT1 appear not to be directly involved in fungicide resistance although can affect the gene expression of fungicide related genes. These results indicate that PdSUT1 contribute to P. digitatum fungal virulence and influence fungicide sensitivity through carbohydrate uptake and MFS transporters gene activation. [ABSTRACT FROM AUTHOR]

Citreoviridin (CIT) is a mycotoxin produced by Penicillum citreonigrum , Aspergillus terreus and Eupenicillium ochrosalmoneum . CIT occurs naturally in moldy rice and corn. CIT is associated with the development of atherosclerosis in the general population. Alteration in hepatic lipid metabolism is a pathogenic factor in atherosclerosis. However the effect and the underlying mechanism of CIT on hepatic lipid metabolism are largely unknown. In this study, we reported that CIT induced triglyceride accumulation in mice liver and human liver HepG2 cells as shown in oil red O staining. CIT (0.1 mg/kg–0.3 mg/kg) for 6 weeks elevated liver triglyceride contents in mice. CIT inhibited the transactivation activity of peroxisome proliferator-activated receptor-α (PPAR-α) in hepatocyte in vivo and in vitro , as shown by the reduced mRNA levels of PPAR-α target genes which play key roles in lipid metabolism in various aspects. PPAR-α agonist fenofibrate attenuated CIT-induced triglyceride accumulation in HepG2 cells. Furthermore, CIT increased serum total cholesterol/high-density lipoprotein cholesterol ratio, a strong risk factor for cardiovascular disease. In summary, we reported that CIT induced PPAR-α−dependent hepatic triglyceride accumulation and dyslipidemia. Our data will provide new mechanistic insights into CIT-induced lipid alterations. [ABSTRACT FROM AUTHOR]

A readily accessible template of 1,5,7-trisubstituted-3-pyridyl-xanthones was designed starting from naturally occurring pyripyropene A for agrichemical development. Our originally developed Ag2CO3-mediated oxidative cyclization enabled ready access to the key scaffold, 1,5,7-trihydroxy-3-chloro-xanthone. The chemo- and regioselective sequential introduction of four substituents to the scaffold rapidly afforded the desired, structurally diverse 1,5,7-trisubstituted-3-pyridyl-xanthones. An evaluation of insecticidal activity revealed that one of the synthesized compounds retained insecticidal activity against vetch aphid and green peach aphid. The observed insecticidal spectrum was similar to that of pyripyropene A. The developed template could be a valuable aid for future agrichemical development. [ABSTRACT FROM AUTHOR]

Kaitocephalin (KCP) isolated from Eupenicillium shearii PF1191 is an unusual amino acid natural product in which serine, proline, and alanine moieties are liked with carbon–carbon bonds. KCP exhibits potent and selective binding affinity for one of the ionotropic glutamate receptor subtypes, NMDA receptors ( K i = 7.8 nM). In this study, new structure–activity relationship studies at C9 of KCP were implemented. Eleven new KCP analogs with different substituents at C9 were prepared and employed for binding affinity tests using native ionotropic glutamate receptors. Replacement of the 3,5-dichloro-4-hydroxybenzoyl group of KCP with a 3-phenylpropionyl group resulted in significant loss of binding affinity for NMDARs ( K i = 1300 nM), indicating an indispensable role of the aromatic ring of KCP in the potent and selective binding to NMDARs. Other analogs showed potent binding affinity in a range of 11–270 nM. These findings would directly link to develop useful chemical tools toward imaging and labeling of NMDARs. [ABSTRACT FROM AUTHOR]

The genus Torulomyces was characterised by species that typically have conidiophores consisting of solitary phialides that produce long chains of conidia connected by disjunctors. Based on the phylogenetic position of P. lagena (generic ex-neotype), the genus and its seven species were transferred to Penicillium and classified in sect. Torulomyces along with P. cryptum and P. lassenii. The aim of this study was to review the species currently classified in sect. Torulomyces using morphology and phylogenies of the ITS, BenA, CaM and RPB2 regions. Based on our results, we accept 16 species in sect. Torulomyces, including 12 new species described as P. aeris, P. austricola, P. cantabricum, P. catalonicum, P. oregonense, P. marthae-christenseniae, P. riverlandense, P. tubakianum, P. variratense, P. williamettense, P. wisconsinense and P. wollemiicola. In addition, we reclassify P. laeve and P. ovatum in sect. Exilicaulis and correct the typification of P. lagena. We provide descriptions and notes on the identification of the species. [ABSTRACT FROM AUTHOR]

Background: Microbial lipids are produced by many oleaginous organisms including the well-characterized yeast Yarrowia lipolytica, which can be engineered for increased lipid yield by up-regulation of the lipid biosynthetic pathway and down-regulation or deletion of competing pathways. Results: We describe a strain engineering strategy centered on diacylglycerol acyltransferase (DGA) gene overexpression that applied combinatorial screening of overexpression and deletion genetic targets to construct a high lipid producing yeast biocatalyst. The resulting strain, NS432, combines overexpression of a heterologous DGA1 enzyme from Rhodosporidium toruloides, a heterlogous DGA2 enzyme from Claviceps purpurea, and deletion of the native TGL3 lipase regulator. These three genetic modifications, selected for their effect on lipid production, enabled a 77 % lipid content and 0.21 g lipid per g glucose yield in batch fermentation. In fed-batch glucose fermentation NS432 produced 85 g/L lipid at a productivity of 0.73 g/L/h. Conclusions: The yields, productivities, and titers reported in this study may further support the applied goal of cost-effective, large -scale microbial lipid production for use as biofuels and biochemicals. [ABSTRACT FROM AUTHOR]

Background: Trichoderma reesei is a widely used model cellulolytic fungus, supplying a highly effective cellulase production system. Recently, the biofuel industry discovered filamentous fungi from the Penicillium genus as a promising alternative to T. reesei. Results: In our study, we present a systematic over-expression analysis of nine β-glucosidase encoding genes in the wild-type strain 114-2 of Penicillium oxalicum. We found that the over-expression of BGL1, BGL4, or BGL5 significantly enhanced both β-glucosidase activity and hydrolysis efficiency of the enzyme system on filter paper. We utilised two strategies to over-express β-glucosidase in the strain RE-10 that--although over-producing cellulase, does so at the cost of the cellulase mixture deficiency. The constitutive promoter of gene pde_02864 encoding 40S ribosomal protein S8 was used to over-express three β-glucosidases: BGL1, BGL4, and BGL5. We found that all mutants show significantly enhanced levels of β-glucosidase at transcriptional, protein, and activity levels. Furthermore, the inducible promoter from bgl2 was used to conditionally over-express the β-glucosidases BGL1 and BGL4. Surprisingly, this induced expression strategy enables significantly improved expression efficiency. The BGL1 over-expressing mutant I1-13 particularly improved the β-glucosidase activity at a factor of 65-folds, resulting in levels of up to 150 U/ml. All our BGL over-expression mutants displayed significant enhancement of cellulolytic ability on both microcrystalline cellulose and filter paper. In addition, they substantially reduced the enzyme loads in the saccharification of a natural lignocellulose material delignified corncob residue (DCCR). The mutant I4-32 with over-expression of BGL4 achieved the highest glucose yield in the saccharification of DCCR at only 25 % enzyme load compared to the parental strain RE-10. Conclusions: In summary, genetically engineering P. oxalicum to significantly improve β-glucosidase activity is a potent strategy to substantially boost the hydrolytic efficiency of the cellulase cocktail, which will ultimately lead to a considerable reduction of cost for biomass-based biofuel. [ABSTRACT FROM AUTHOR]

Phenalenones are polyketide natural products that display diverse structures and biological activities. The core of phenalenones is a peri-fused tricyclic ring system cyclized from a linear polyketide precursor via an unresolved mechanism. Toward understanding the unusual cyclization steps, the phn biosynthetic gene cluster responsible for herqueinone biosynthesis was identified from the genome of Penicillium herquei. A nonreducing polyketide synthase (NR-PKS) PhnA was shown to synthesize the heptaketide backbone and cyclize it into the angular, hemiketal-containing naphtho-γ-pyrone prephenalenone. The product template (PT) domain of PhnA catalyzes only the C4-C9 aldol condensation, which is unprecedented among known PT domains. The transformation of prephenalenone to phenalenone requires an FAD-dependent monooxygenase (FMO) PhnB, which catalyzes the C2 aromatic hydroxylation of prephenalenone and ring opening of the γ-pyrone ring simultaneously. Density functional theory calculations provide insights into why the hydroxylated intermediate undergoes an aldol-like phenoxide-ketone cyclization to yield the phenalenone core. This study therefore unveiled new routes and biocatalysts for polyketide cyclization. [ABSTRACT FROM AUTHOR]

The aim of the research was a comparative study of the thermal stability of native and modified by various methods α-L rhamnosidases of Eupenicillium erubescens and Cryptococcus albidus for improving the stability of enzymes. Denaturation of native and modified enzymes were performed at 65 0C, pH 5.2. Enzyme activity was determined using p-nitrophenyl-α-L-rhamnopyranoside and naringin. It is found that in the treatment by polyethylenglycol PEG 1500, dextrans 500 and 70 T thermostability of α-L-rhamnosidases tested decreases, while modification with polyethylenglycol 20000 leads to increase thermal stability of the E. erubescens enzyme to 280% and C. albidus to 150%. Comparative study of the thermal stability of the native and modified by cellulose and its derivatives α-L-rhamnosidases of C. albidus and E. erubescens showed that at concentrations cellulose of 5-15 μg/10 μg protein protective effect of polymers on enzymes was investigated was observed. Hydrophobic modifications using succinic anhydride also can slow down the denaturation of α-L-rhamnosidases tested under experimental conditions. These stabilized C. albidus and E. erubescens α-L-rhamnosidases can be used in biotechnological processes. [ABSTRACT FROM AUTHOR]

Preparation of inoculum of biocontrol species in the most suitable carrier material is imperative to get full benefit from its resistance induction potential. Present study focuses upon the evaluation of supportive behavior of carrier materials towards Penicillium oxalicum Currie & Thom in controlling Alternaria leaf spots of tomato caused by Alternaria alternata (Fr.) Keissl. Inoculum of resistance inducer species was prepared in five different carrier materials namely talc, bentonite, sand, saw dust and Bhagaas; and incubated at room temperature for 0, 10, 20×80 days. Then soil applications of all those treatments under the field conditions revealed that Bhagaas based inducer inoculum with no incubation period controlled fungal disease most efficiently. Furthermore, evaluation of agro-economic aspects of tomato plants also proved Bhagaas as the best carrier material by which high yield and maximum vigor index was achieved. Second most efficient carrier was saw dust followed by talc, sand and bentonite, respectively. Present study concludes that inoculum of P. oxalicum should be prepared using Bhagaas as cerrier material. [ABSTRACT FROM AUTHOR]

The total synthesis of epoxyeujindole A, a structurally unusual indole diterpenoid isolated from Eupenicillium javanicum, has been accomplished for the first time. The synthesis features a late-stage cationic cyclization strategy, which took advantage of an electron-rich olefinic substrate. The CDE ring system was assembled via an enantioselective conjugate addition/alkylation, a Luche cyclization, and a Nozaki-Hiyama-Kishi reaction. The heavily substituted A ring was constructed through a Suzuki-Miyaura coupling and a cationic cyclization, and the bridged fused B ring was formed through a Prins reaction. [ABSTRACT FROM AUTHOR]

The recent discovery and functional characterization of opposite mating-type loci in the industrial penicillin producer Penicillium chrysogenum demonstrated their regulatory role in sexual as well as asexual development. Subsequent experiments further showed that a sexual life cycle can be induced in P. chrysogenum that was for long believed to reproduce exclusively by asexual propagation. Finally, crossing of wild type and production strains resulted in the generation of recombinant ascospore isolates. We predict from these recent findings that recombinant progeny for industrial applications can be obtained by sexual crossings and discuss experimental difficulties that occur when parental strains with karyotype heterogeneity are used for mating. [ABSTRACT FROM AUTHOR]

Recombinant strains of Penicillium canescens producing homologous pectin lyase A and heterologous endo-1,5-α-arabinase A and endo-1,4-α-polygalacturonase, as well as enzymes of the host strain (α-L-arabinofuranosidases, xylanases, and others), were obtained by genetic engineering. The enzyme preparations (EPs) obtained from the cultural medium of recombinant P. canescens strains efficiently hydrolyzed raw plant material with a high content of pectin compounds. It was shown that the yield of reducing sugars and arabinose increased 16 and 22% in comparison with the control EP based on the host strain when one of the obtained EPs was used for beet pulp hydrolysis. It was established that the most active EP consisted of pectin lyase (10%), endo-1,5-arabinase (26%),α-L-arabinofuranosidase and arabinoxylan-arabinofuranohydrolase (12%), and xylanase (10%). The activities of pectin lyase, polygalacturonase, and arabinase of the EP in reactions with various substrates were determined. The specificity, pH and T-optima, and thermal stability of the homogenous recombinant endo-1,5-α-arabinase were investigated. The kinetic parameters ( K, k) of the linear arabinan hydrolysis were determined. [ABSTRACT FROM AUTHOR]

A phytase from Penicillium oxalicum PJ3, PhyA, was purified near to homogeneity with 427-fold increase in specific phytase activity by ammonium sulfate precipitation, gel filtration, and ion-exchange chromatographies. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and zymogram analysis of the purified enzyme indicated an estimated molecular mass of 65 kD. The optimal pH and temperature of the purified enzyme were pH 4.5 and 55°C, respectively. The enzyme activity was strongly inhibited by Ca2+, Cu2+, Zn2+, and phenylmethylsulfonyl fluoride (PMSF). The Km value for sodium phytate was 0.545 mM with a Vmax of 600 U/mg of protein. The phyA gene was cloned, and it contains an open reading frame of 1,383 with a single intron (118 bp), and encodes a protein of 461 amino acids. [ABSTRACT FROM AUTHOR]

The FAOD/FPOD family of proteins has the potential to be useful for the longterm detection of blood glucose levels in diabetes patients. A bottleneck for this application is to find or engineer a FAOD/FPOD family enzyme that is specifically active towards α-fructosyl peptides but is inactive towards other types of glycated peptides. Here, the crystal structure of fructosyl peptide oxidase from Eupenicillium terrenum (EtFPOX) is reported at 1.9 Å resolution. In contrast to the previously reported structure of amadoriase II, EtFPOX has an open substrate entrance to accommodate the large peptide substrate. The functions of residues critical for substrate selection are discussed based on structure comparison and sequence alignment. This study reveals the first structural details of group I FPODs that prefer α-fructosyl substrates and could provide significant useful information for uncovering the mechanism of substrate specificity of FAOD/FPODs and guidance towards future enzyme engineering for diagnostic purposes. [ABSTRACT FROM AUTHOR]

In this work, a biosensor was constructed by physical adsorption of the isolated endophytic fungus Eupenicillium shearii FREI-39 esterase on halloysite, using graphite powder, multi-walled carbon nanotubes and mineral oil for the determination of carbofuran pesticide by inhibition of the esterase using square-wave voltammetry (SWV). Specific esterase activities were determined each 2 days over a period of 15 days of growth in four different inoculation media. The highest specific activity was found on 6th day, with 33.08 U on PDA broth. The best performance of the proposed biosensor was obtained using 0.5 U esterase activity. The carbofuran concentration response was linear in the range from 5.0 to 100.0 µg L −1 ( r =0.9986) with detection and quantification limits of 1.69 µg L −1 and 5.13 µg L −1 , respectively. A recovery study of carbofuran in spiked water samples showed values ranging from 103.8±6.7% to 106.7±9.7%. The biosensor showed good repeatability and reproducibility and remained stable for a period of 20 weeks. The determination of carbofuran in spiked water samples using the proposed biosensor was satisfactory when compared to the chromatographic reference method. The results showed no significant difference at the 95% confidence level with t -test statistics. The application of enzymes from endophytic fungi in constructing biosensors broadens the biotechnological importance of these microorganisms. [ABSTRACT FROM AUTHOR]

Extracellular alkalinization and HO production are important early events during induced systemic resistance (ISR) establishment in plants. In a screen for metabolites as potential ISR activators from 98 fungal isolates associated with marine sponge Hymeniacidon perleve, the crude metabolites of fungus Eupenicillium brefeldianum HMP-F96 induced significant extracellular alkalinization coupled with HO production in tobacco cell suspensions. A combined bioactivity and H NMR-guided fractionation approach was used to disclose the chemical determinants responsible for the activities. Eight cyclic dipeptides were purified from the fermentation broth of the strain and were structurally characterized by NMR and MS experiments. This study represents the first report of the occurrence of cyclic dipeptides in E. brefeldianum and of their activities of inducing extracellular alkalinization and HO production in tobacco cell suspensions. [ABSTRACT FROM AUTHOR]

Biocatalysts with lipase activity were produced by Penicillium corylophilum under basal and olive oil-induced conditions. In the presence of olive oil, both the extracellular and the mycelium-bound lipase activities were increased significantly. The four biocatalysts maintained a residual lipase activity after incubation in a wide pH range showing high stability in a defined acidic to neutral pH range. Some differences between biocatalyst residual activities were also observed after incubation at various temperatures or in the presence of organic solvents such as methanol, ethanol, acetone, butanol, hexanol, n-hexane, and heptane. Enzymes which are stable in the presence of ethanol or butanol could be useful in ecofriendly processes for biodiesel production. [ABSTRACT FROM AUTHOR]

A major challenge for further promotion of lipase productivity in Penicillium expansum PE-12 is to find a suitable promoter that can function efficiently in this industrial strain. In this study, the 5′ flanking region of P. expansum lipase ( Ppel) containing a putative novel promoter sequence was characterized by fusing to [ABSTRACT FROM AUTHOR]

The ethyl acetate extracts of the culture of Eupenicillium parvum, Gelasinospora brevispora, Neosartoryap seudofischeri, N. quadricincta and N. multiplicata were assessed for their antifungal activity against ten economically important plant pathogenic fungi: Pythiumaphanidermatum, Phytophthorapalmivora, Alternaria sp., Fusariumoxysporum, Colletotrichumgloeosporioides, Lasiodiplodiatheobromae, Helminthosporiummaydis, Sclerotiumrolfsii and Rhizoctoniasolani, which arecausative agents of fruit and vegetable diseases. The bioassay for the antifungal activity of the fungal crude extracts was based on the dilution plate method. Although all the extracts exhibited a complete inhibition of the mycelial growth of some plant pathogenic fungi at the highest concentration tested (10,000 ppm), an interesting antifungal effect was observed for the crude extract of N. pseudofischeri KUFA 0060 against Ph. palmivora and C. capsiciat 100 ppm, as well as for N. quadricincta KUFA 0064 against Ph. palmivora and Alternaria sp. at 10 ppm. The results obtained from this screening allow us to identify new potential sources for the development of alternative fungicides. [ABSTRACT FROM AUTHOR]

Literature on the hyphomycete genus Penicillium Link and its teleomorphs, Eupenicillium Ludwig and Talaromyces C.R.Benjamin, is surveyed in the Republic of South Africa, Lesotho, Mozambique, Namibia, Swaziland and Transkei up to 1990. References are grouped under the headings, general mycology, plant pathology, industrial application, medical importance, mycotoxins and chemical work. An alphabetical list of the species recorded in southern Africa as well as the host and/or substrate from which each species has been reported is presented with relevant literature references; specimens in various culture collections are also incorporated. Although most of the known Penicillium species have already been reported from southern Africa, in-depth work is still required in all fields of research concerning this genus.

Moldy Civil is a mold-ripened variety of cheese produced mainly in eastern Turkey. This cheese is produced with Civil cheese and whey curd cheese (Lor). Civil cheese has had a geographical presence since 2009 and is manufactured with skim milk. In the production of Moldy Civil cheese, Civil cheese or a mixture of Civil and Lor cheese is pressed into goat skins or plastic bags and ripened for 3 months or longer. During the ripening period, natural contaminating molds grow on the surface of and inside the cheese. In this study, 186 mold strains were isolated from 41 samples of Moldy Civil cheese, and 165 of these strains were identified as Penicillium roqueforti. Identification and mycotoxicologic analyses were conducted using morphotypic and molecular methods. PCR amplicons of the ITS1-5.SS-ITS4 region were subjected to sequence analysis. This research is the first using molecular methods on Moldy Civil cheese. Mycotoxicologic analyses were conducted using thin-layer chromatography, and random amplified polymorphic DNA genotypes were determined using the aril primer. Of 165 isolates, only 28 produced no penicillic acid, P. roqueforti toxin, or roquefortine. [ABSTRACT FROM AUTHOR]

In this work, a robust brefeldin A-synthesizing fungus, Eupenicillium brefeldianum ZJB082702, was bred from a Murraya paniculata endophytic fungus E. brefeldianum A1163. Using one-factor-at-a-time experimental design, optimization of media composition for E. brefeldianum ZJB082702 fermenting brefeldin A was conducted. Outcomes indicated that mixed carbon source and mixed nitrogen source were of c ritical importance to brefeldin A fermentation. After 6d culture in the optimized fermentation media, composed of (gl−1) 13.33 starch, 26.67 glucose, 1.0 yeast extract powder, 1.0 corn steep liquor, 0.5 soybean meal, 0.75 NaNO3, 2.5 malt extract, 6.0 CaCO3, 3.0 MgSO4, 4.0 KH2PO4, 1.0 × 10−2 CuSO4, brefeldin A yield peaked at 1304.7mgl−1, 648.2mgl−1 in 500ml baffled flask and 15l stirred fermentor respectively, formed as a growth associated type of secondary metabolite based on fermentation profile analysis. [Copyright &y& Elsevier]

Abstract: Brefeldin A (BFA) is a macrolide lactone antibiotic, possessing antitumor, antiviral, antifungal activities. In this work, a separation strategy involving one-step macroporous resin adsorption chromatography combined with crystallization was established for BFA purification from Eupenicillium brefeldianum CCTCC M 208113 fermentation broth. Among six macroporous resin adsorbents tested, the non-polar resin HZ830 had the best adsorption and desorption performance. The static equilibrium adsorption data fitted well with the Freundlich equation, and the adsorption kinetic followed the pseudo-second order model. Through experimental optimization of column adsorption and desorption, BFA in purity of 90.4% (w/w), 92.1% (w/w) yield was obtained by a one-step macroporous resin adsorption chromatography, using a stepwise elution protocol. Furthermore, high purity (>99%, w/w) of BFA crystals were prepared from E. brefeldianum CCTCC M 208113 fermentation broth in an overall recovery of 67.0% (w/w), using a combination of adsorption chromatography packed with non-polar macroporous adsorbent HZ830 and crystallization in acetone. [Copyright &y& Elsevier]

Twenty-six samples of spices most frequently occurring in the stores of the Republic of Macedonia were examined for their fungal contamination and the incidence of Aspergillus and Penicillium species and their teleomorphs. It included mainly commercial packages most frequently occurring in the stores of the Republic of Macedonia. According to the relative frequency of each of the isolated species, the typical mycoflora of these samples includes A. niger, A. flavus, A. fumigatus, P. chrysogenum and Eurotium sp. Fungal counts varied from log10 < 2 CFU g-1 (DG18 at 25° C; DRBC at 25°C), for a sample of paprika, to log10 6.17 CFU g-1 (DG18 at 25°C), for a sample of bay leaf. During the experiment, A. flavus was detected in 17 samples, out of which 7 isolates were capable of producing Af-B1, and 4 isolates produced Af-B2. All isolates of A. nomius and A. parasiticus, in the experimental conditions, produced Af-B1, Af-B22, Af-G1 and Af-G2. [ABSTRACT FROM AUTHOR]

Abstract: Eupenicillium species are the teleomorphic (sexual) forms of anamorphic (asexual) members of the genus Penicillium, which contains many species of industrial importance. Here we describe the first molecular analysis of the mating-type (MAT) locus from a homothallic (self-fertile) Eupenicillium species, E. crustaceum. This ascomycete is a sexual relative of the penicillin producer Penicillium chrysogenum, which while long considered asexual, was recently shown to possess the required genetic machinery for heterothallic breeding. The E. crustaceum genome contains two MAT loci, MAT1-1 and MAT1-2, in an arrangement characteristic of other known homothallic euascomycetes, such as Neosartorya fischeri. MAT1-1 is flanked by conserved APN2 (DNA lyase) and SLA2 (cytoskeleton assembly control) genes and encodes a homologue of the α-box domain protein MAT1-1-1. Conversely, MAT1-2 carries a HMG-domain gene MAT1-2-1, and is flanked by a degenerate SLA2 gene and an intact homologue of the P. chrysogenum ORF Pc20g08960. Here we demonstrate the transcriptional expression of both mating-type genes during vegetative development. Furthermore, the MAT1-1-1 and MAT1-2-1 sequences were used to resolve the phylogenetic relationship of E. crustaceum with other ascomycetes. Phylogenetic trees confirmed a very close relationship between the homothallic E. crustaceum and the supposedly heterothallic P. chrysogenum. This close taxonomic association makes E. crustaceum an ideal candidate for future expression and evolutionary studies of sexual reproduction, with the ultimate aim of inducing sex in P. chrysogenum. [Copyright &y& Elsevier]

Bingtang sweet orange processing waste was utilised to produce four feed enzymes (Endoglucanase, β-glucosidase, pectinase and xylanase) by the solid-state fermentation (SSF) with Eupenicillium javanicum. The factors related with SSF including moisture content, temperature, initial pH, time, carbon source (0.5 g), nitrogen sources (0.05 g), inorganic mineral salts (0.1 g) were investigated separately. The corresponding optimal condition was: moisture content 80% (w/w), temperature 30 °C, natural pH, time 96 h, wheat bran 0.5 g, (NH)SO 0.05 g or NaNO 0.05 g, CaCl 0.1 g. The L(3) orthogonal experiment results showed that the optimal condition for producing above multiple enzymes was: moisture content 80% (w/w), temperature 30 °C, wheat bran 1 g, (NH)SO 0.05 g, NaNO 0.05 g, CaCl 0.1 g, fermentation time 96 h and natural pH. Under this condition, the average activity of Endoglucanase (CMCase), β-glucosidase, pectinase and xylanase by E. javanicum could reach 46.80, 49.64, 51.87 and 106.42 U g, respectively, which was significantly higher than those in single factor experiments. Our present results demonstrated that E. javanicum could also be an effective and useful fungus for multienzyme preparation especially for β-glucosidase and xylanase from citrus processing wastes. [ABSTRACT FROM AUTHOR]

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Abstract: Dextransucrase from Leuconostoc mesenteroides and dextranase from Penicillium lilacinum were co-immobilized and used to produce isomaltooligosaccharides from sucrose. The enzymes were co-immobilized by encapsulating soluble dextransucrase and dextranase covalently attached to Eupergit C in alginate (beads, fibers, and capsules). The alginate capsule co-immobilization was done in the presence of soluble starch and resulted in a high immobilization yield (71%), and the enzymes retained their activities during 20 repeated batch reactions and for a month in storage at 4°C. The presence of starch was essential for the stability of dextransucrase in alginate capsules. Furthermore, it is important that the dextranase be pre-immobilized prior to alginate capsule co-immobilization to prevent dextranase leakage and inactivation of dextransucrase. The co-immobilized enzymes formed oligosaccharides from sucrose, which can be used as prebiotics. In addition, the oligosaccharides that were produced after the addition of sucrose reacted with the alginate fiber-encapsulted dextransucrase, thus increasing the amount of prebiotics. Co-immobilization in alginate fiber and beads also resulted in high yields (70 and 64%), but enzymatic activities decreased by 74 and 99%, respectively, after a month in storage at 4°C. The newly developed alginate capsule method for co-immobilization of dextransucrase and dextranase is simple yet effective and has the potential for industrial-scale production of isomaltooligosaccharides. [ABSTRACT FROM AUTHOR]

Abstract: A mature phytase cDNA, encoding 441 amino acids, from Eupenicillium parvum (BCC17694) was cloned into a Pichia pastoris expression vector, pPICZαA, and was successfully expressed as active extracellular glycosylated protein. The recombinant phytase contained the active site RHGXRXP and HD sequence motifs, a large α/β domain and a small α-domain that are typical of histidine acid phosphatase. Glycosylation was found to be important for enzyme activity which is most active at 50°C and pH 5.5. The recombinant phytase displayed broad substrate specificity toward p-nitrophenyl phosphate, sodium-, calcium-, and potassium-phytate. The enzyme lost its activity after incubating at 50°C for 5min and is 50% inhibited by 5mM Cu2+. However, the enzyme exhibits broad pH stability from 2.5 to 8.0 and is resistant to pepsin. In vitro digestibility test suggested that BCC17694 phytase is at least as effective as another recombinant phytase (r-A170) which is comparable to Natuphos, a commercial phytase, in releasing phosphate from corn-based animal feed, suggesting that BCC17694 phytase is suitable for use as phytase supplement in the animal diet. [Copyright &y& Elsevier]

Abstract: Herbimycin A, a typically water-insoluble anti-tumor drug and an analog of geldanamycin, was transformed by fungus Eupenicillium sp. SD017. The water solubility and anti-tumor activity of converted products were evaluated after separation, purification, and structure elucidation. The results showed that herbimycin A can be converted into herbimycin C 1 and 11-hydroxy-(11-demethoxy)-herbimycin C 2 with water solubility increased. The latter is a novel compound, and its IC50 to A549, MCF-7, Ehrlich and HeLa cells is 152.9, 0.46, 10.7 and 109.3μg/ml, respectively, which are nearly in the range of those between herbimycin A and herbimycin C. It is suggested that an O-demethylase or/and methyltransferase may play an important role on biotransformation in this case. This study will firstly serve as a fundamental methodology for further structural alteration and functional improvement of the water-insoluble anti-tumor drugs using microbial resources. [Copyright &y& Elsevier]

Studies on production of secondary metabolites by fungi have received a substantial boost lately, particularly with reference to applications of their biological properties in human medicine. Funicones represent a series of related compounds for which there is accumulating evidence supporting their possible use as pharmaceuticals. This paper provides a review on the current status of knowledge on these fungal extrolites, with special reference to aspects concerning their molecular structures and biological activities. [ABSTRACT FROM AUTHOR]