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Sample records for penicillium corylophilum dierckx

  1. Biodegradation of glyceryl trinitrate by Penicillium corylophilum Dierckx.

    PubMed Central

    Zhang, Y Z; Sundaram, S T; Sharma, A; Brodman, B W

    1997-01-01

    Penicillium corylophilum Dierckx, isolated from a contaminated water wet, double-base propellant, was able to completely degrade glyceryl trinitrate (GTN) in a buffered medium (pH 7.0) containing glucose and ammonium nitrate. In the presence of 12 mg of initial fungal inoculum, GTN (48.5 to 61.6 mumol) was quantitatively transformed in a stepwise process to glyceryl dinitrate (GDN) and glyceryl mononitrate (GMN) within 48 h followed by a decrease in the GDN content with a concomitant increase in the GMN level. GDN was totally transformed to GMN within 168 h, and the complete degradation of GMN was achieved within 336 h. The presence of glucose and ammonium nitrate in the growth medium was essential for completion of the degradation of GTN and its metabolites. Complete degradation of GTN by a fungal culture has not been previously reported in the literature. PMID:9143106

  2. Secondary metabolites from Penicillium corylophilum isolated from damp buildings.

    PubMed

    McMullin, David R; Nsiama, Tienabe K; Miller, J David

    2014-01-01

    Indoor exposure to the spores and mycelial fragments of fungi that grow on damp building materials can result in increased non-atopic asthma and upper respiratory disease. The mechanism appears to involve exposure to low doses of fungal metabolites. Penicillium corylophilum is surprisingly common in damp buildings in USA, Canada and western Europe. We examined isolates of P. corylophilum geographically distributed across Canada in the first comprehensive study of secondary metabolites of this fungus. The sesquiterpene phomenone, the meroterpenoids citreohybridonol and andrastin A, koninginin A, E and G, three new alpha pyrones and four new isochromans were identified from extracts of culture filtrates. This is the first report of koninginins, meroterpenoids and alpha pyrones from P. corylophilum. These secondary metabolite data support the removal of P. corylophilum from Penicillium section Citrina and suggest that further taxonomic studies are required on this species.

  3. Identification of citrinin as the defence metabolite of Penicillium corylophilum stressed with the antagonist fungus Beauveria bassiana.

    PubMed

    Dos Santos, Claudia Maria Campinha; da Costa, Gisela Lara; Figueroa-Villar, José Daniel

    2012-01-01

    Penicillium corylophilum isolated from mosquitoes was cultivated in liquid media leading to the first reported identification of citrinin (1a) as one metabolic component of this fungus. The produced amount of 1a indicated this compound as the most abundant secondary metabolite of this fungus. Stressing the culture of P. corylophilum with the presence of the antagonistic fungus Beauveria bassiana increased in 65% the production of 1a. Similar results were obtained with the presence of other fungi in the culture media, indicating that citrinin is the main defence metabolite of P. corylophilum. In agreement with this conclusion, citrinin showed a reasonable fungicidal activity against Colletotrichum gloeosporioides and B. bassiana.

  4. Entomopathogenic effect of Aspergillus giganteus and Penicillium corylophilum on two triatomine vectors of Chagas disease.

    PubMed

    da Costa, Gisela Lara; de Moraes, Aurea Maria Lage; Galvão, Cleber

    2003-01-01

    Two strains, Penicillium corylophilum and Aspergillus giganteus, of the most frequent species found in a survey of triatomines, were used for bioassays in the second and fourth nymphs stage of Triatoma infestans and Panstrongylus megistus. Two procedures, bite and pulverization, were used and compared. A. giganteus was most effective, causing mortality in at least 50% of the nymphs of the two species tested with exception of the nymphs of the fourth stage of P. megistus. Variation in entomopathogenic capacity of the fungal species were observed in the experiments. The two procedures used proved effective.

  5. Inflammatory and cytotoxic responses in mouse lungs exposed to purified toxins from building isolated Penicillium brevicompactum Dierckx and P. chrysogenum Thom.

    PubMed

    Rand, Thomas G; Giles, S; Flemming, J; Miller, J David; Puniani, Eva

    2005-09-01

    In vitro and in vivo studies have shown that building-associated Penicillium spores and spore extracts can induce significant inflammatory responses in lung cells and animal models of lung disease. However, because spores and spore extracts comprise mixtures of bioactive constituents often including toxins, it is impossible to resolve which constituent mediates inflammatory responses. This study examined dose-response (0.5 nM, 2.5 nM, 5.0 nM, 12.5 nM/g body weight (BW) animal) and time-course (3, 6, 24 and 48 h post instillation (PI)) relationships associated with inflammatory and cytotoxic responses in mouse lungs intratracheally instilled with pure brevianamide A, mycophenolic acid, and roquefortine C. High doses (5.0 nM and/or 12.5 nM/g BW animal) of brevianamide A and mycophenolic acid, the dominant metabolites of P. brevicompactum, and roquefortine C, the dominant metabolite of P. chrysogenum, induced significant inflammatory responses within 6 h PI, expressed as differentially elevated macrophage, neutrophil, MIP-2, TNF, and IL-6 concentrations in the bronchioalveolar lavage fluid (BALF) of intratracheally exposed mice. Macrophage and neutrophil numbers were maximal at 24 h PI; responses of the other inflammatory markers were maximal at 6 h PI. Except for macrophage numbers in mycophenolic acid-treatment animals, cells exhibited significant dose-dependent-like responses; for the chemo-/cytokine markers, dose dependency was lacking except for MIP-2 concentration in brevianamide A-treatment animals. It was also found that brevianamide A induced cytotoxicity expressed as significantly increased LDH concentration in mouse BALF, at concentrations of 12.5 nM/g BW animal and at 6 and 24 h PI. Albumin concentrations, measured as a nonspecific marker of vascular leakage, were significantly elevated in the BALF of mice treated with 12.5 nM/g nM brevianamide A/animal from 6 to 24 h PI and in > or =5.0 nM/g mycophenolic acid-treated animals at 6 to 24 h PI. These results

  6. Effect of water activity and temperature on growth of three Penicillium species and Aspergillus flavus on a sponge cake analogue.

    PubMed

    Abellana, M; Sanchis, V; Ramos, A J

    2001-12-30

    This study compared the effect of temperature and water activity and their interactions on the rate of mycelial growth of Penicillium aurantiogriseum, P. chrysogenum, P. corylophilum and Aspergillus flavus on a sponge cake analogue. As expected, growth rates showed dependence on a(w), and temperature. However, no significant differences were observed in the growth rates of different isolates. The minimum a(w) values for growth of the Penicillium spp. was 0.85-0.90. A. flavus was able to grow at 0.90 a(w) when the temperature was above 15 degrees C. This study has shown that fungal growth by these species on a sponge cake analogue, with a composition similar to usual bakery products, is prevented if the a(w) is kept at < 0.85.

  7. Phylogeny and nomenclature of the genus Talaromyces and taxa accommodated in Penicillium subgenus Biverticillium

    PubMed Central

    Samson, R.A.; Yilmaz, N.; Houbraken, J.; Spierenburg, H.; Seifert, K.A.; Peterson, S.W.; Varga, J.; Frisvad, J.C.

    2011-01-01

    The taxonomic history of anamorphic species attributed to Penicillium subgenus Biverticillium is reviewed, along with evidence supporting their relationship with teleomorphic species classified in Talaromyces. To supplement previous conclusions based on ITS, SSU and/or LSU sequencing that Talaromyces and subgenus Biverticillium comprise a monophyletic group that is distinct from Penicillium at the generic level, the phylogenetic relationships of these two groups with other genera of Trichocomaceae was further studied by sequencing a part of the RPB1 (RNA polymerase II largest subunit) gene. Talaromyces species and most species of Penicillium subgenus Biverticillium sensu Pitt reside in a monophyletic clade distant from species of other subgenera of Penicillium. For detailed phylogenetic analysis of species relationships, the ITS region (incl. 5.8S nrDNA) was sequenced for the available type strains and/or representative isolates of Talaromyces and related biverticillate anamorphic species. Extrolite profiles were compiled for all type strains and many supplementary cultures. All evidence supports our conclusions that Penicillium subgenus Biverticillium is distinct from other subgenera in Penicillium and should be taxonomically unified with the Talaromyces species that reside in the same clade. Following the concepts of nomenclatural priority and single name nomenclature, we transfer all accepted species of Penicillium subgenus Biverticillium to Talaromyces. A holomorphic generic diagnosis for the expanded concept of Talaromyces, including teleomorph and anamorph characters, is provided. A list of accepted Talaromyces names and newly combined Penicillium names is given. Species of biotechnological and medical importance, such as P. funiculosum and P. marneffei, are now combined in Talaromyces. Excluded species and taxa that need further taxonomic study are discussed. An appendix lists other generic names, usually considered synonyms of Penicillium sensu lato that

  8. Two new Penicillium species Penicillium buchwaldii and Penicillium spathulatum, producing the anticancer compound asperphenamate.

    PubMed

    Frisvad, Jens C; Houbraken, Jos; Popma, Suuske; Samson, Robert A

    2013-02-01

    Penicillium buchwaldii sp. nov. (type strain CBS 117181(T)  = IBT 6005(T)  = IMI 30428(T) ) and Penicillium spathulatum sp. nov. (CBS 117192(T)  = IBT 22220(T) ) are described as new species based on a polyphasic taxonomic approach. Isolates of P. buchwaldii typically have terverticillate conidiophores with echinulate thick-walled conidia and produce the extrolites asperphenamate, citreoisocoumarin, communesin A and B, asperentin and 5'-hydroxy-asperentin. Penicillium spathulatum is unique in having restricted colonies on Czapek yeast agar (CYA) with an olive grey reverse, good growth on CYA supplemented with 5% NaCl, terverticillate bi- and ter-ramulate conidiophores and consistently produces the extrolites benzomalvin A and D and asperphenamate. The two new species belong to Penicillium section Brevicompacta and are phylogenetically closely related to Penicillium tularense. With exception of Penicillium fennelliae, asperphenamate is also produced by all other species in section Brevicompacta (P. tularense, Penicillium brevicompactum, Penicillium bialowiezense, Penicillium olsonii, Penicillium astrolabium and Penicillium neocrassum). Both new species have a worldwide distribution. The new species were mainly isolated from indoor environments and food and feedstuffs. The fact that asperphenamate has been found in many widely different plants may indicate that endophytic fungi rather than the plants are the actual producers.

  9. Penicillium viridicatum, Penicillium verrucosum, and production of ochratoxin A.

    PubMed Central

    Pitt, J I

    1987-01-01

    The taxonomy of the important mycotoxigenic species Penicillium viridicatum and P. verrucosum was reviewed to clarify disagreements relating to the three P. viridicatum groups erected by Ciegler and coworkers (A. Ciegler, D. I. Fennell, G. A. Sansing, R. W. Detroy, and G. A. Bennett, Appl. Microbiol. 26:271-278, 1973) and the mycotoxins produced by them. Cultures derived from the types of these two species and authentic cultures from each group and from many other sources were examined culturally, microscopically, and for mycotoxin production. It was concluded that P. viridicatum group II has affinities with P. verrucosum and not with P. viridicatum, as indicated by J. I. Pitt in the 1979 monograph (The Genus Penicillium and Its Teleomorphic States Eupenicillium and Talaromyces). As a result of this study it can now be unequivocally stated that the mycotoxins ochratoxin A and citrinin are not produced by P. viridicatum. Of species in subgenus Penicillium, only P. verrucosum is known to produce ochratoxin A. PMID:3566267

  10. Fructooligosaccharide production by Penicillium expansum.

    PubMed

    Prata, Margarida B; Mussatto, Solange I; Rodrigues, Lígia R; Teixeira, José A

    2010-06-01

    Fructooligosaccharides (FOS) production by Penicillium expansum was evaluated. In a first stage, the best conditions for P. expansum growth and sporulation were established with potato/dextrose/agar being the most suitable medium at between 22 and 25 degrees C, giving good growth and good sporulation. The inocula from this medium were used for FOS production using shake-flask cultures, and yielded 0.58 g FOS/g sucrose (3.25 g FOS/l.h), demonstrating the potential of this strain for sucrose conversion to FOS.

  11. Taxonomy of Penicillium section Citrina

    PubMed Central

    Houbraken, J.; Frisvad, J.C.; Samson, R.A.

    2011-01-01

    Species of Penicillium section Citrina have a worldwide distribution and occur commonly in soils. The section is here delimited using a combination of phenotypic characters and sequences of the nuclear ribosomal RNA gene operon, including the internal transcribed spacer regions ITS1 and ITS2, the 5.8S nrDNA (ITS) and partial RPB2 sequences. Species assigned to section Citrina share the production of symmetrically biverticillate conidiophores, flask shaped phialides (7.0–9.0 μm long) and relatively small conidia (2.0–3.0 μm diam). Some species can produce greyish-brown coloured cleistothecia containing flanged ascospores. In the present study, more than 250 isolates presumably belonging to section Citrina were examined using a combined analysis of phenotypic and physiological characters, extrolite profiles and ITS, β-tubulin and/or calmodulin sequences. Section Citrina includes 39 species, and 17 of those are described here as new. The most important phenotypic characters for distinguishing species are growth rates and colony reverse colours on the agar media CYA, MEA and YES; shape, size and ornamentation of conidia and the production of sclerotia or cleistothecia. Temperature-growth profiles were made for all examined species and are a valuable character characters for species identification. Species centered around P. citrinum generally have a higher maximum growth temperature (33–36 °C) than species related to P. westlingii (27–33 °C). Extrolite patterns and partial calmodulin and β-tubulin sequences can be used for sequence based identification and resolved all species. In contrast, ITS sequences were less variable and only 55 % of the species could be unambiguously identified with this locus. Taxonomic novelties: Penicillium argentinense Houbraken, Frisvad & Samson, P. atrofulvum Houbraken, Frisvad & Samson, P. aurantiacobrunneum Houbraken, Frisvad & Samson, P. cairnsense Houbraken, Frisvad & Samson, P. christenseniae Houbraken, Frisvad & Samson

  12. Identification and nomenclature of the genus Penicillium.

    PubMed

    Visagie, C M; Houbraken, J; Frisvad, J C; Hong, S-B; Klaassen, C H W; Perrone, G; Seifert, K A; Varga, J; Yaguchi, T; Samson, R A

    2014-06-01

    Penicillium is a diverse genus occurring worldwide and its species play important roles as decomposers of organic materials and cause destructive rots in the food industry where they produce a wide range of mycotoxins. Other species are considered enzyme factories or are common indoor air allergens. Although DNA sequences are essential for robust identification of Penicillium species, there is currently no comprehensive, verified reference database for the genus. To coincide with the move to one fungus one name in the International Code of Nomenclature for algae, fungi and plants, the generic concept of Penicillium was re-defined to accommodate species from other genera, such as Chromocleista, Eladia, Eupenicillium, Torulomyces and Thysanophora, which together comprise a large monophyletic clade. As a result of this, and the many new species described in recent years, it was necessary to update the list of accepted species in Penicillium. The genus currently contains 354 accepted species, including new combinations for Aspergillus crystallinus, A. malodoratus and A. paradoxus, which belong to Penicillium section Paradoxa. To add to the taxonomic value of the list, we also provide information on each accepted species MycoBank number, living ex-type strains and provide GenBank accession numbers to ITS, β-tubulin, calmodulin and RPB2 sequences, thereby supplying a verified set of sequences for each species of the genus. In addition to the nomenclatural list, we recommend a standard working method for species descriptions and identifications to be adopted by laboratories working on this genus.

  13. [HIV-associated Penicillium marneffei infection].

    PubMed

    Kronauer, C M; Schär, G; Barben, M; Bühler, H

    1993-03-06

    We report on an HIV positive patient with a disseminated Penicillium marneffei infection. A 35-year-old Swiss homosexual male with HIV-associated immunodeficiency with a CD4 cell count of 90/mm3 presented with a two-month history of malaise, intermittent fever, loss of weight, unproductive cough and widespread molluscum contagiosum-like skin lesions, mainly on the face. The patient had travelled extensively and had last visited Thailand 19 months before admission. The chest X-ray showed bilateral diffuse reticulonodular markings. The diagnosis was suspected in bronchoalveolar lavage, which showed round-to-oval intracellular yeast cells but also elongated sausage-shaped extracellular forms. The diagnosis was confirmed on culture. Penicillium marneffei was further isolated from the following specimens: blood cultures, bone marrow, stool, skin and tracheal mucosa biopsy. Intravenous amphotericin B therapy led to a complete subsidence of all symptoms and the skin lesions healed without leaving a scar. The infection, with its clinical presentation, epidemiology, diagnostic problems and therapy is reviewed. We stress that since Penicillium marneffei is an increasingly important pathogen in HIV positive patients in Southeast Asia, this fungus can also be imported to Europe by travellers. If immunocompromised patients have molluscum contagiosum-like skin lesions, pneumonitis and a history of travelling in Southeast Asia, disseminated Penicillium marneffei infection should be considered in differential diagnosis.

  14. Identification and nomenclature of the genus Penicillium

    PubMed Central

    Visagie, C.M.; Houbraken, J.; Frisvad, J.C.; Hong, S.-B.; Klaassen, C.H.W.; Perrone, G.; Seifert, K.A.; Varga, J.; Yaguchi, T.; Samson, R.A.

    2014-01-01

    Penicillium is a diverse genus occurring worldwide and its species play important roles as decomposers of organic materials and cause destructive rots in the food industry where they produce a wide range of mycotoxins. Other species are considered enzyme factories or are common indoor air allergens. Although DNA sequences are essential for robust identification of Penicillium species, there is currently no comprehensive, verified reference database for the genus. To coincide with the move to one fungus one name in the International Code of Nomenclature for algae, fungi and plants, the generic concept of Penicillium was re-defined to accommodate species from other genera, such as Chromocleista, Eladia, Eupenicillium, Torulomyces and Thysanophora, which together comprise a large monophyletic clade. As a result of this, and the many new species described in recent years, it was necessary to update the list of accepted species in Penicillium. The genus currently contains 354 accepted species, including new combinations for Aspergillus crystallinus, A. malodoratus and A. paradoxus, which belong to Penicillium section Paradoxa. To add to the taxonomic value of the list, we also provide information on each accepted species MycoBank number, living ex-type strains and provide GenBank accession numbers to ITS, β-tubulin, calmodulin and RPB2 sequences, thereby supplying a verified set of sequences for each species of the genus. In addition to the nomenclatural list, we recommend a standard working method for species descriptions and identifications to be adopted by laboratories working on this genus. PMID:25505353

  15. Detection of Extracellular Enzyme Activity in Penicillium using Chromogenic Media.

    PubMed

    Yoon, Ji Hwan; Hong, Seung Beom; Ko, Seung Ju; Kim, Seong Hwan

    2007-09-01

    A total of 106 Penicillium species were tested to examine their ability of degrading cellobiose, pectin and xylan. The activity of β-glucosidase was generally strong in all the Penicillium species tested. P. citrinum, P. charlesii, P. manginii and P. aurantiacum showed the higher ability of producing β-glucosidase than other tested species. Pectinase activity was detected in 24 Penicillium species. P. paracanescens, P. sizovae, P. sartoryi, P. chrysogenum, and P. claviforme showed strong pectinase activity. In xylanase assay, 84 Penicillium species showed activity. Strong xylanase activity was detected from P. megasporum, P. sartoryi, P. chrysogenum, P. glandicola, P. discolor, and P. coprophilum. Overall, most of the Penicillium species tested showed strong β-glucosidase activity. The degree of pectinase and xylanase activity varied depending on Penicillium species.

  16. Diclavatol and tetronic acids from Penicillium griseoroseum.

    PubMed

    da Silva, José Vinicius; Fill, Taicia Pacheco; da Silva, Bianca Ferreira; Rodrigues-Fo, Edson

    2013-01-01

    In our continuous studies on the chemistry of the endophytic fungus Penicillium griseoroseum, an endophyte isolated from fruits of Coffea arabica, we isolated clavatol, a dimethylated tetraketide, and its dimer which appears to be a novel natural compound. The studies also resulted in the identification of two known tetronic acids, viridicatic acid and terrestric acid, found in ethyl acetate and n-butanol extracts. Spectroscopic studies using 1-D and 2-D NMR and MS/MS analysis were performed to determine the structures of these compounds, first reported by this Penicillium. Two other tetronic acids congeners were identified through HPLC/MS/MS studies, based on fragmentation pattern of ions produced from ionised tetronic acids, and UV light absorptions.

  17. Expanding the species and chemical diversity of Penicillium section Cinnamopurpurea

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A set of isolates very similar to or potentially conspecific with an unidentified Penicillium isolate NRRL 735, was assembled using a BLAST search of ITS similarity among described (GenBank) and undescribed Penicillium isolates in our laboratories. DNA was amplified from six loci of the assembled is...

  18. Expanding the species and chemical diversity of Penicillium section Cinnamopurpurea

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A set of isolates genetically similar to or potentially conspecific with an unidentified Penicillium isolate NRRL 735, was assembled using a Basic Local Alignment Search Tool (BLAST) search of internal transcribed spacer (ITS) similarity among described (GenBank) and undescribed Penicillium isolates...

  19. Draft Genome Sequence of the Fungus Penicillium brasilianum MG11

    PubMed Central

    Linde, Jörg; Mattern, Derek J.; Walther, Grit; Guthke, Reinhard; Brakhage, Axel A.

    2015-01-01

    The genus Penicillium belongs to the phylum Ascomycota and includes a variety of fungal species important for food and drug production. We report the draft genome sequence of Penicillium brasilianum MG11. This strain was isolated from soil, and it was reported to produce different secondary metabolites. PMID:26337871

  20. Non-gravitational effects on genus penicillium

    NASA Technical Reports Server (NTRS)

    Loup, Mackenzie

    1995-01-01

    In September 1994, Shuttle Orbiter Discovery, STS-64, launched into space. Aboard that shuttle was a payload containing Fungi spores, genus Penicillium. With the over looking help of Dr. Audrey Gabel, Associate Professor of Biology at Black Hills State University, investigations on differing media types began. Basis for this experimentation was to determine if there was any differences between the space exposed spores and control spores. Studies concluded that there were differences and those differences were then recorded. It was hypothesized the spores may have been effected causing differences in growth rate, colony size, depth and margins, coloring, germination, and growth on different media.

  1. Non-gravitational effects on genus penicillium

    SciTech Connect

    Loup, M.

    1995-09-01

    In September 1994, Shuttle Orbiter Discovery, STS-64, launched into space. Aboard that shuttle was a payload containing Fungi spores, genus Penicillium. With the over looking help of Dr. Audrey Gabel, Associate Professor of Biology at Black Hills State University, investigations on differing media types began. Basis for this experimentation was to determine if there was any differences between the space exposed spores and control spores. Studies concluded that there were differences and those differences were then recorded. It was hypothesized the spores may have been effected causing differences in growth rate, colony size, depth and margins, coloring, germination, and growth on different media.

  2. Species-specific identification of penicillium linked to patulin contamination.

    PubMed

    Dombrink-Kurtzman, Mary Ann; McGovern, Amy E

    2007-11-01

    Certain species of Penicillium have been reported to produce the mycotoxin patulin, and research was undertaken to identify these with the use of oligonucleotide primer pairs. Species examined were found in food, plants, and soil and were reported to produce patulin. Penicillium expansum is the most commonly detected species linked to the presence of patulin in apple juice. At least 10 different enzymes are involved in the patulin biosynthetic pathway, including the isoepoxydon dehydrogenase (idh) gene. Based on nucleotide sequences previously determined for the idh gene in Penicillium species, PCR primers were designed for the species-specific detection of patulin-producing species. The 5' primers were based on differences in the second intron of the idh gene. To ensure that the primer pairs produced a PCR product restricted to the species for which it was designed, and not to unrelated species, all of the primer pairs were tested against all of the Penicillium species. With one exception, it was possible to detect a reaction only with the organism of interest. The primer pair for Penicillium griseofulvum also amplified DNA from Penicillium dipodomyicola, a closely related species; however, it was possible to distinguish between these two species by doing a second amplification, with a different primer pair specific only for P. dipodomyicola. Consequently, with different primer sets, it was possible to identify individual patulin-producing species of Penicillium.

  3. Hydrolysis of lignocelluloses by penicillium funiculosum cellulase

    SciTech Connect

    Mishra, C.; Rao, M.; Seeta, R.; Srinivasan, M.C.; Deshpande, V.

    1984-04-01

    Enzymatic hydrolysis of cellulose is a promising method for the conversion of waste cellulose to glucose. During the past few years, the development of this technology has proceeded rapidly, with significant advances made in enzyme production, pretreatment, and hydrolysis. A variety of fungi are reported to produce cellulases but among these Trichoderma reesei and its mutants are powerful producers of cellulases. However, the search for new and possibly better sources of cellulase is continued due to the low levels of beta-glucosidase of T. reesei. Penicillium funiculosum produces a complete cellulase having endo-beta-1,4-glucanase (15-20 U/mL), exo-beta-1,4-glucanase (1.5-2.0 U/mL), and high beta-glucosidase (8-10 U/mL). The saccharification of alkali-treated cotton and bagasse by P. funiculosum enzyme was 70 and 63%, respectively. It was possible to obtain glucose concentration as high as 30% using 50% bagasse. It is of interest that the percent saccharification of cellulosic substrates with the Penicillium enzyme is comparable to that of T. reesei cellulase when the same amount of filter paper activity is used, although the endo-glucanase activity of the latter is two to three times higher. This communication reports the studies on saccharification of lignocelluloses by P. funiculosum cellulase and certain studies on the kinetic aspects. (Refs. 15).

  4. Individual and combined effects of vanillin and potassium sorbate on Penicillium digitatum, Penicillium glabrum, and Penicillium italicum growth.

    PubMed

    Matamoros-León, B; Argaiz, A; López-Malo, A

    1999-05-01

    The individual and combined effects of potassium sorbate and vanillin concentrations on the growth of Penicillium digitatum, P. glabrum, and P. italicum in potato dextrose agar adjusted to water activity 0.98 and pH 3.5 were evaluated. Inhibitory concentrations of potassium sorbate varied from 150 ppm for P. digitatum to 700 ppm for P. glabrum, and for vanillin from 1,100 ppm for P. digitatum and P. italicum and 1,300 ppm for P. glabrum. Fractional inhibitory concentration (FIC) isobolograms show curves deviated to the left of the additive line. Calculated FIC index varied from 0.60 to 0.84. FIC index as well as FIC isobolograms show synergistic effects on mold inhibition when vanillin and potassium sorbate are applied in combination.

  5. Ultraviolet Radiation Induction of Mutation in Penicillium Claviforme.

    ERIC Educational Resources Information Center

    New, June; Jolley, Ray

    1986-01-01

    Cites reasons why Penicillium claviforme is an exceptionally good species for ultraviolet induced mutation experiments. Provides a set of laboratory instructions for teachers and students. Includes a discussion section. (ML)

  6. Pyran Rings Containing Polyketides from Penicillium raistrickii

    PubMed Central

    Ma, Li-Ying; Liu, De-Sheng; Li, De-Guo; Huang, Yu-Ling; Kang, Hui-Hui; Wang, Chun-Hua; Liu, Wei-Zhong

    2016-01-01

    Five new pyran rings containing polyketides, penicipyrans A–E (1–5), together with the known pestapyrone A (6), were isolated from the saline soil-derived Penicillium raistrickii. Their structures were determined by interpretation of NMR and HRESIMS data. The absolute configurations of compounds 4 and 5 were established by the modified Mosher’s method and single-crystal X-ray diffraction analysis, respectively. These compounds possessed high structural diversity including two α-pyrones (1, 2), three isocoumarins (3, 4, 6), and one dihydropyran derivative (5). Among them, Compound 5 exhibited cytotoxicity against HL-60 and K562 cell lines with IC50 values of 4.4 and 8.5 μM, respectively. PMID:28025533

  7. Production of Patulin by Penicillium expansum

    PubMed Central

    Sommer, Noel F.; Buchanan, Jack R.; Fortlage, Robert J.

    1974-01-01

    Twenty-seven isolates of Penicillium expansum Lk. ex Thom obtained from Europe, Australia, and North America from seven different fruit hosts all produced patulin in culture. Six isolates were essentially nonpathogenic in apple fruits. In culture, patulin generally accumulated to much higher levels than in apple fruits. At all temperatures permitting fungus growth, patulin was produced. However, only small amounts were observed near the maximal temperature for growth (30 C). At 0 C, patulin accumulated but slowly in culture. Modified atmospheres suppressed both fungus growth and patulin accumulation in apples. After varying incubation periods to obtain similar total growth, the patulin concentration was low in modified atmospheres and high in air. PMID:4425020

  8. Contrasting Genomic Diversity in Two Closely Related Postharvest Pathogens: Penicillium digitatum and Penicillium expansum.

    PubMed

    Julca, Irene; Droby, Samir; Sela, Noa; Marcet-Houben, Marina; Gabaldón, Toni

    2015-12-14

    Penicillium digitatum and Penicillium expansum are two closely related fungal plant pathogens causing green and blue mold in harvested fruit, respectively. The two species differ in their host specificity, being P. digitatum restricted to citrus fruits and P. expansum able to infect a wide range of fruits after harvest. Although host-specific Penicillium species have been found to have a smaller gene content, it is so far unclear whether these different host specificities impact genome variation at the intraspecific level. Here we assessed genome variation across four P. digitatum and seven P. expansum isolates from geographically distant regions. Our results show very high similarity (average 0.06 SNPs [single nucleotide polymorphism] per kb) between globally distributed isolates of P. digitatum pointing to a recent expansion of a single lineage. This low level of genetic variation found in our samples contrasts with the higher genetic variability observed in the similarly distributed P. expansum isolates (2.44 SNPs per kb). Patterns of polymorphism in P. expansum indicate that recombination exists between genetically diverged strains. Consistent with the existence of sexual recombination and heterothallism, which was unknown for this species, we identified the two alternative mating types in different P. expansum isolates. Patterns of polymorphism in P. digitatum indicate a recent clonal population expansion of a single lineage that has reached worldwide distribution. We suggest that the contrasting patterns of genomic variation between the two species reflect underlying differences in population dynamics related with host specificities and related agricultural practices. It should be noted, however, that this results should be confirmed with a larger sampling of strains, as new strains may broaden the diversity so far found in P. digitatum.

  9. Functional characterization of Penicillium occitanis Pol6 and Penicillium funiculosum GH11 xylanases.

    PubMed

    Driss, Dorra; Berrin, Jean Guy; Juge, Nathalie; Bhiri, Fatma; Ghorbel, Raoudha; Chaabouni, Semia Ellouz

    2013-08-01

    Xylanases are hemicellulolytic enzymes, which are responsible for the degradation of heteroxylans constituting the lignocellulosic plant cell wall. Xylanases from the GH11 family are considered as true xylanases because of their high substrate specificity. In order to study in depth a crucial difference in the thumb region between two closely related xylanases from Penicillium in terms of kinetic parameters and inhibition sensitivity, the GH11 xylanases from Penicillium occitanis Pol6 (PoXyn3) and from Penicillium funiculosum (PfXynC) were heterologously expressed in Pichia pastoris. The PoXyn3 and PfXynC cDNAs encoding mature xylanases were cloned into pGAPZαA vectors and integrated into the genome of P. pastoris X-33 under the control of the glyceraldehyde 3-phosphate dehydrogenase constitutive promoter. PfXynC was expressed as a His-tagged recombinant protein and purified from the supernatant homogeneity by a one-step purification protocol using immobilized metal affinity chromatography. The recombinant PoXyn3 was purified using a single anion-exchange chromatography. The purified recombinant enzymes were optimally active at 45°C and pH 4.0 for PoXyn3 and 40°C and pH 3.0 for PfXynC. The measured kinetic parameters (k(cat) and Vmax) showed that PfXynC was five times more active than PoXyn3 irrespective of the substrate whereas the apparent affinity (K(m)) was similar. The recombinant enzymes showed distinct sensitivity to the Triticum aestivum xylanase inhibitor TAXI-I.

  10. Contrasting Genomic Diversity in Two Closely Related Postharvest Pathogens: Penicillium digitatum and Penicillium expansum

    PubMed Central

    Julca, Irene; Droby, Samir; Sela, Noa; Marcet-Houben, Marina; Gabaldón, Toni

    2016-01-01

    Penicillium digitatum and Penicillium expansum are two closely related fungal plant pathogens causing green and blue mold in harvested fruit, respectively. The two species differ in their host specificity, being P. digitatum restricted to citrus fruits and P. expansum able to infect a wide range of fruits after harvest. Although host-specific Penicillium species have been found to have a smaller gene content, it is so far unclear whether these different host specificities impact genome variation at the intraspecific level. Here we assessed genome variation across four P. digitatum and seven P. expansum isolates from geographically distant regions. Our results show very high similarity (average 0.06 SNPs [single nucleotide polymorphism] per kb) between globally distributed isolates of P. digitatum pointing to a recent expansion of a single lineage. This low level of genetic variation found in our samples contrasts with the higher genetic variability observed in the similarly distributed P. expansum isolates (2.44 SNPs per kb). Patterns of polymorphism in P. expansum indicate that recombination exists between genetically diverged strains. Consistent with the existence of sexual recombination and heterothallism, which was unknown for this species, we identified the two alternative mating types in different P. expansum isolates. Patterns of polymorphism in P. digitatum indicate a recent clonal population expansion of a single lineage that has reached worldwide distribution. We suggest that the contrasting patterns of genomic variation between the two species reflect underlying differences in population dynamics related with host specificities and related agricultural practices. It should be noted, however, that this results should be confirmed with a larger sampling of strains, as new strains may broaden the diversity so far found in P. digitatum. PMID:26672008

  11. Modern taxonomy of biotechnologically important Aspergillus and Penicillium species.

    PubMed

    Houbraken, Jos; de Vries, Ronald P; Samson, Robert A

    2014-01-01

    Taxonomy is a dynamic discipline and name changes of fungi with biotechnological, industrial, or medical importance are often difficult to understand for researchers in the applied field. Species belonging to the genera Aspergillus and Penicillium are commonly used or isolated, and inadequate taxonomy or uncertain nomenclature of these genera can therefore lead to tremendous confusion. Misidentification of strains used in biotechnology can be traced back to (1) recent changes in nomenclature, (2) new taxonomic insights, including description of new species, and/or (3) incorrect identifications. Changes in the recent published International Code of Nomenclature for Algae, Fungi and Plants will lead to numerous name changes of existing Aspergillus and Penicillium species and an overview of the current names of biotechnological important species is given. Furthermore, in (biotechnological) literature old and invalid names are still used, such as Aspergillus awamori, A. foetidus, A. kawachii, Talaromyces emersonii, Acremonium cellulolyticus, and Penicillium funiculosum. An overview of these and other species with their correct names is presented. Furthermore, the biotechnologically important species Talaromyces thermophilus is here combined in Thermomyces as Th. dupontii. The importance of Aspergillus, Penicillium, and related genera is also illustrated by the high number of undertaken genome sequencing projects. A number of these strains are incorrectly identified or atypical strains are selected for these projects. Recommendations for correct strain selection are given here. Phylogenetic analysis shows a close relationship between the genome-sequenced strains of Aspergillus, Penicillium, and Monascus. Talaromyces stipitatus and T. marneffei (syn. Penicillium marneffei) are closely related to Thermomyces lanuginosus and Th. dupontii (syn. Talaromyces thermophilus), and these species appear to be distantly related to Aspergillus and Penicillium. In the last part of

  12. Polyketides, toxins and pigments in Penicillium marneffei.

    PubMed

    Tam, Emily W T; Tsang, Chi-Ching; Lau, Susanna K P; Woo, Patrick C Y

    2015-10-30

    Penicillium marneffei (synonym: Talaromyces marneffei) is the most important pathogenic thermally dimorphic fungus in China and Southeastern Asia. The HIV/AIDS pandemic, particularly in China and other Southeast Asian countries, has led to the emergence of P. marneffei infection as an important AIDS-defining condition. Recently, we published the genome sequence of P. marneffei. In the P. marneffei genome, 23 polyketide synthase genes and two polyketide synthase-non-ribosomal peptide synthase hybrid genes were identified. This number is much higher than those of Coccidioides immitis and Histoplasma capsulatum, important pathogenic thermally dimorphic fungi in the Western world. Phylogenetically, these polyketide synthase genes were distributed evenly with their counterparts found in Aspergillus species and other fungi, suggesting that polyketide synthases in P. marneffei did not diverge from lineage-specific gene duplication through a recent expansion. Gene knockdown experiments and ultra-high performance liquid chromatography-photodiode array detector/electrospray ionization-quadruple time of flight-mass spectrometry analysis confirmed that at least four of the polyketide synthase genes were involved in the biosynthesis of various pigments in P. marneffei, including melanin, mitorubrinic acid, mitorubrinol, monascorubrin, rubropunctatin, citrinin and ankaflavin, some of which were mycotoxins and virulence factors of the fungus.

  13. Reminiscence of phospholipase B in Penicillium notatum.

    PubMed

    Saito, Kunihiko

    2014-01-01

    Since the phospholipase B (PLB) was reported as a deacylase of both lecithin and lysolecithin yielding fatty acids and glycerophosphocholine (GPC), there was a question as to whether it is a single enzyme or a mixture of a phospholipase A2 (PLA2) and a lysophospholipase (LPL). We purified the PLB in Penicillium notatum and showed that it catalyzed deacylation of sn-1 and sn-2 fatty acids of 1,2-diacylphospholipids and also sn-1 or sn-2 fatty acids of 1- or 2-monoacylphospholipids (lysophospholipids). Further, it also has a monoacyllipase activity. The purified PLB is a glycoprotein with m.w. of 91,300. The sugar moiety is M9 only and the protein moiety consists of 603 amino acids. PLB, different from PLA2, shows other enzymatic activities, such as transacylase, lipase and acylesterase. PLB activity is influenced by various substances, e.g. detergents, deoxycholate, diethylether, Fe(3+), and endogenous protease. Therefore, PLB might have broader roles than PLA2 in vivo. The database shows an extensive sequence similarity between P. notatum PLB and fungal PLB, cPLA2 and patatin, suggesting a homologous relationship. The catalytic triad of cPLA2, Ser, Asp and Arg, is also present in P. notatum PLB. Other related PLBs, PLB/Lipases are discussed.

  14. Effect of Penicillium chrysogenum on Lignin Transformation

    PubMed Central

    Rodríguez, A.; Carnicero, A.; Perestelo, F.; de la Fuente, G.; Milstein, O.; Falcón, M. A.

    1994-01-01

    A strain of Penicillium chrysogenum has been isolated from pine forest soils in Tenerife (Canary Islands). This strain was capable of utilizing hydroxylated and nonhydroxylated aromatic compounds, in particular cinnamic acid, as its sole carbon source. In an optimum medium with high levels of nitrogen (25.6 mM) and low levels of glucose (5.5 mM), it was able to decolorize Poly B-411 and to transform kraft, organosolv, and synthetic dehydrogenative polymerisate lignins. After 30 days of incubation, the amount of recovered kraft lignin was reduced to 83.5 and 91.3% of that estimated for uninoculated controls by spectrophotometry and klason lignin, respectively. At the same time, the pattern of molecular mass distribution of the lignin remaining in cultures was changed. The amount of organosolv lignin recovered from cultures was reduced to 90.1 and 94.6% of the initial amount as evaluated by spectrophotometry and klason lignin, respectively. About 6% of total applied radioactivity of O14CH3-organosolv lignin was recovered as 14CO2 after 30 days of incubation, and 18.5% of radioactivity from insoluble O14CH3-organosolv lignin was solubilized. After 26 days of incubation, 2.9% of 14C-β-dehydrogenative polymerisate and 4.1% of 14C-ring-dehydrogenative polymerisate evolved as 14CO2. PMID:16349361

  15. Penicillium roqueforti PR toxin gene cluster characterization.

    PubMed

    Hidalgo, Pedro I; Poirier, Elisabeth; Ullán, Ricardo V; Piqueras, Justine; Meslet-Cladière, Laurence; Coton, Emmanuel; Coton, Monika

    2017-03-01

    PR toxin is a well-known isoprenoid mycotoxin almost solely produced by Penicillium roqueforti after growth on food or animal feed. This mycotoxin has been described as the most toxic produced by this species. In this study, an in silico analysis allowed identifying for the first time a 22.4-kb biosynthetic gene cluster involved in PR toxin biosynthesis in P. roqueforti. The pathway contains 11 open reading frames encoding for ten putative proteins including the major fungal terpene cyclase, aristolochene synthase, involved in the first farnesyl-diphosphate cyclization step as well as an oxidoreductase, an oxidase, two P450 monooxygenases, a transferase, and two dehydrogenase enzymes. Gene silencing was used to study three genes (ORF5, ORF6, and ORF8 encoding for an acetyltransferase and two P450 monooxygenases, respectively) and resulted in 20 to 40% PR toxin production reductions in all transformants proving the involvement of these genes and the corresponding enzyme activities in PR toxin biosynthesis. According to the considered silenced gene target, eremofortin A and B productions were also affected suggesting their involvement as biosynthetic intermediates in this pathway. A PR toxin biosynthesis pathway is proposed based on the most recent and available data.

  16. Functional diversity within the Penicillium roqueforti species.

    PubMed

    Gillot, Guillaume; Jany, Jean-Luc; Poirier, Elisabeth; Maillard, Marie-Bernadette; Debaets, Stella; Thierry, Anne; Coton, Emmanuel; Coton, Monika

    2017-01-16

    Penicillium roqueforti is used as a ripening culture for blue cheeses and largely contributes to their organoleptic quality and typical characteristics. Different types of blue cheeses are manufactured and consumed worldwide and have distinct aspects, textures, flavors and colors. These features are well accepted to be due to the different manufacturing methods but also to the specific P. roqueforti strains used. Indeed, inoculated P. roqueforti strains, via their proteolytic and lipolytic activities, have an effect on both blue cheese texture and flavor. In particular, P. roqueforti produces a wide range of flavor compounds and variations in their proportions influence the flavor profiles of this type of cheese. Moreover, P. roqueforti is also characterized by substantial morphological and genetic diversity thus raising the question about the functional diversity of this species. In this context, 55 representative strains were screened for key metabolic properties including proteolytic activity (by determining free NH2 amino groups) and secondary metabolite production (aroma compounds using HS-Trap GC-MS and mycotoxins via LC-MS/Q-TOF). Mini model cheeses were used for aroma production and proteolysis analyses, whereas Yeast Extract Sucrose (YES) agar medium was used for mycotoxin production. Overall, this study highlighted high functional diversity among isolates. Noteworthy, when only P. roqueforti strains isolated from Protected Designation of Origin (PDO) or Protected Geographical Indication (PGI) blue cheeses were considered, a clear relationship between genetic diversity, population structure and the assessed functional traits was shown.

  17. Polyketides, Toxins and Pigments in Penicillium marneffei

    PubMed Central

    Tam, Emily W. T.; Tsang, Chi-Ching; Lau, Susanna K. P.; Woo, Patrick C. Y.

    2015-01-01

    Penicillium marneffei (synonym: Talaromyces marneffei) is the most important pathogenic thermally dimorphic fungus in China and Southeastern Asia. The HIV/AIDS pandemic, particularly in China and other Southeast Asian countries, has led to the emergence of P. marneffei infection as an important AIDS-defining condition. Recently, we published the genome sequence of P. marneffei. In the P. marneffei genome, 23 polyketide synthase genes and two polyketide synthase-non-ribosomal peptide synthase hybrid genes were identified. This number is much higher than those of Coccidioides immitis and Histoplasma capsulatum, important pathogenic thermally dimorphic fungi in the Western world. Phylogenetically, these polyketide synthase genes were distributed evenly with their counterparts found in Aspergillus species and other fungi, suggesting that polyketide synthases in P. marneffei did not diverge from lineage-specific gene duplication through a recent expansion. Gene knockdown experiments and ultra-high performance liquid chromatography-photodiode array detector/electrospray ionization-quadruple time of flight-mass spectrometry analysis confirmed that at least four of the polyketide synthase genes were involved in the biosynthesis of various pigments in P. marneffei, including melanin, mitorubrinic acid, mitorubrinol, monascorubrin, rubropunctatin, citrinin and ankaflavin, some of which were mycotoxins and virulence factors of the fungus. PMID:26529013

  18. Infection capacities in the orange-pathogen relationship: compatible (Penicillium digitatum) and incompatible (Penicillium expansum) interactions.

    PubMed

    Vilanova, L; Viñas, I; Torres, R; Usall, J; Jauset, A M; Teixidó, N

    2012-02-01

    Penicillium digitatum and Penicillium expansum are the most devastating pathogens of citrus and pome fruits, respectively. Whereas P. digitatum is a very specific pathogen that only infects Citrus fruits, P. expansum has a broader host range but has not been reported to be infectious in Citrus. To determine the responses of fruits and the infection capacities of both moulds, two varieties of oranges at different maturity stages, different inoculum concentrations and two different storage temperatures were studied. In compatible interactions, no significant differences in rot dynamics among harvests were found with a 10(7) conidia mL(-1) inoculum concentration at both temperatures tested (20 °C and 4 °C). However, at other inoculum concentrations, significant differences in rot dynamics were found, especially in immature fruits. Incompatible interactions showed that P. expansum could infect oranges at commercial maturity in both tested varieties. Decay incidence and severity were higher at 4 °C than at 20 °C. In addition to infection capacity studies, histochemical tests were performed to detect wound-healing compounds for both pathogens. A positive reaction for lignin was detected for both pathogens in immature oranges over a short period (48 h). In all cases, no reactions were found in control samples. Our results indicate that pathogen concentration, host maturity and storage temperature can play important roles in the defence mechanisms of fruit. Furthermore, to our knowledge, this is the first work that demonstrates that P. expansum can infect oranges under favourable conditions.

  19. Effect of LED Blue Light on Penicillium digitatum and Penicillium italicum Strains.

    PubMed

    Lafuente, María T; Alférez, Fernando

    2015-11-01

    Studies on the antimicrobial properties of light have considerably increased due in part to the development of resistance to actual control methods. This study investigates the potential of light-emitting diodes (LED) blue light for controlling Penicillium digitatum and Penicillium italicum. These fungi are the most devastating postharvest pathogens of citrus fruit and cause important losses due to contaminations and the development of resistant strains against fungicides. The effect of different periods and quantum fluxes, delaying light application on the growth and morphology of P. digitatum strains resistant and sensitive to fungicides, and P. italicum cultured at 20°C was examined. Results showed that blue light controls the growth of all strains and that its efficacy increases with the quantum flux. Spore germination was always avoided by exposing the cultures to high quantum flux (700 μmol m(-2) s(-1) ) for 18 h. Continuous light had an important impact on the fungus morphology and a fungicidal effect when applied at a lower quantum flux (120 μmol m(-2) s(-1) ) to a growing fungus. Sensitivity to light increased with mycelium age. Results show that blue light may be a tool for P. digitatum and P. italicum infection prevention during handling of citrus fruits.

  20. First morphomolecular identification of Penicillium griseofulvum and Penicillium aurantiogriseum toxicogenic isolates associated with blue mold on apple.

    PubMed

    Moslem, Mohmed; Abd-Elsalam, Kamel; Yassin, Mohamed; Bahkali, Ali

    2010-07-01

    Postharvest blue mold decay caused by Penicillium spp. is the most important disease of fresh apple fruit in the world, which extend from the field to the store. Two new Penicillium spp. responsible for apple fruit decay were recovered. The morphological and molecular features of Penicillium griseofulvum and Penicillium aurantiogriseum isolated from apple fruits were characterized morphologically and molecularly. Pathogenicity test exhibited that both P. griseofulvum and P. aurantiogriseum were responsible for blue mold decay in storage apple fruits. Lesion diameter indicated that P. aurantiogriseum was more aggressive than P. griseofulvum. All tested isolates were able to synthesize citrinin in addition to patulin. Not all of the isolates belonging to the same species showed the same profile of secondary metabolites. Microsatellite-primed polymerase chain reaction was able to differentiate these isolates at the species level and divided the analyzed isolates into two genetically different groups. Little intraspecific variability was evident. Microsatellite-primed polymerase chain reaction analysis proved to be an objective, rapid, and reliable tool to identify Penicillium spp. involved in blue mold of apple. This is the first report of occurrence of P. griseofulvum and P. aurantiogriseum on imported apple fruits in Saudi Arabia.

  1. Antifungal Activity of Eugenol against Penicillium, Aspergillus, and Fusarium Species.

    PubMed

    Campaniello, Daniela; Corbo, Maria Rosaria; Sinigaglia, Milena

    2010-06-01

    The antifungal activity of eugenol in a model system against aspergilli (Aspergillus niger, Aspergillus terreus, and Emericella nidulans), penicilli (Penicillium expansum, Penicillium glabrum, and Penicillium italicum), and fusaria (Fusarium oxysporum and Fusarium avenaceum) was investigated. Minimum detection time (time to attain a colony diameter of 1 cm) and the kinetic parameters were evaluated. The effectiveness of the active compound seemed to be strain or genus dependent; 100 mg/liter represented a critical value for P. expansum, P. glabrum, P. italicum, A. niger, and E. nidulans because a further increase of eugenol resulted in fungistatic activity. The radial growth of A. terreus and F. avenaceum was inhibited at 140 mg/liter, and growth of F. oxysporum was completely inhibited at 150 mg/liter.

  2. Penicillium koreense sp. nov., isolated from various soils in Korea.

    PubMed

    You, Young-Hyun; Cho, Hye Sun; Song, Jaekyeong; Kim, Dae-Ho; Houbraken, Jos; Hong, Seung-Beom

    2014-12-28

    During an investigation of the fungal diversity of Korean soils, four Penicillium strains could not be assigned to any described species. The strains formed monoverticillate conidiophores with occasionally a divaricate branch. The conidia were smooth or finely rough-walled, globose to broadly ellipsoidal and 2.5-3.5 × 2.0-3.0 μm in size. Their taxonomic novelty was determined using partial β-tubulin gene sequences and the ribosomal internal transcribed spacer region. The phylogenetic analysis showed that the isolates belonged to section Lanata- Divaricata and were most closely related to Penicillium raperi. Phenotypically, the strains differed from P. raperi in having longer and thicker stipes and thicker phialides. Strain KACC 47721(T) from bamboo field soil was designated as the type strain of the new species, and the species was named Penicillium koreense sp. nov., as it was isolated from various regions in Korea.

  3. Toxigenic Aspergillus and Penicillium isolates from weevil-damaged chestnuts.

    PubMed

    Wells, J M; Payne, J A

    1975-10-01

    Aspergillus and Penicillium were among the most common genera of fungi isolated on malt-salt agar from weevil-damaged Chinese chestnut kernels (16.8 and 40.7% occurrence, respectively). Chloroform extracts of 21 of 50 Aspergillus isolates and 18 of 50 representative Penicillium isolates, grown for 4 weeks at 21.1 C on artificial medium, were toxic to day-old cockerels. Tweleve of the toxic Aspergillus isolates were identified as A. wentii, eight as A. flavus, and one as A. flavus var. columnaris. Nine of the toxic Penicillium isolates were identified as P. terrestre, three as P. steckii, two each as P. citrinum and P. funiculosum, and one each as P. herquei (Series) and P. roqueforti (Series). Acute diarrhea was associated with the toxicity of A. wentii and muscular tremors with the toxicity of P. terrestre, one isolate of P. steckii, and one of P. funiculosum.

  4. Novel Penicillium cellulases for total hydrolysis of lignocellulosics.

    PubMed

    Marjamaa, Kaisa; Toth, Karolina; Bromann, Paul Andrew; Szakacs, George; Kruus, Kristiina

    2013-05-10

    The (hemi)cellulolytic systems of two novel lignocellulolytic Penicillium strains (Penicillium pulvillorum TUB F-2220 and P. cf. simplicissimum TUB F-2378) have been studied. The cultures of the Penicillium strains were characterized by high cellulase and β-glucosidase as well moderate xylanase activities compared to the Trichoderma reesei reference strains QM 6a and RUTC30 (volumetric or per secreted protein, respectively). Comparison of the novel Penicillium and T. reesei secreted enzyme mixtures in the hydrolysis of (ligno)cellulose substrates showed that the F-2220 enzyme mixture gave higher yields in the hydrolysis of crystalline cellulose (Avicel) and similar yields in hydrolysis of pre-treated spruce and wheat straw than enzyme mixture secreted by the T. reesei reference strain. The sensitivity of the Penicillium cellulase complexes to softwood (spruce) and grass (wheat straw) lignins was lignin and temperature dependent: inhibition of cellulose hydrolysis in the presence of wheat straw lignin was minor at 35°C while at 45°C by spruce lignin a clear inhibition was observed. The two main proteins in the F-2220 (hemi)cellulase complex were partially purified and identified by peptide sequence similarity as glycosyl hydrolases (cellobiohydrolases) of families 7 and 6. Adsorption of the GH7 enzyme PpCBH1 on cellulose and lignins was studied showing that the lignin adsorption of the enzyme is temperature and pH dependent. The ppcbh1 coding sequence was obtained using PCR cloning and the translated amino acid sequence of PpCBH1 showed up to 82% amino acid sequence identity to known Penicillium cellobiohydrolases.

  5. Three new records of penicillium species isolated from insect specimens in Korea.

    PubMed

    Lamsal, Kabir; Kim, Sang Woo; Naeimi, Shahram; Adhikari, Mahesh; Yadav, Dil Raj; Kim, Changmu; Lee, Hyang Burm; Lee, Youn Su

    2013-06-01

    Three Penicillium species have been isolated from insect specimens in Korea; Penicillium sp., P. steckii, and P. polonicum. Penicillium sp. (KNU12-3-2) was isolated from Lixus imperessiventris, while P. polonicum (KNU12-1-8) and Penicillium steckii (KNU12-2-9) were isolated from Muljarus japonicas and Meloe proscarabaeus, respectively. The identification was based on the morphological characteristics of the fungi and in internal transcribed spacer analysis. This is the first report on the isolation of these three species of Penicillium from insects in Korea.

  6. Visual clone identification of Penicillium commune isolates.

    PubMed

    Hansen, Michael Edberg; Lund, Flemming; Carstensen, Jens Michael

    2003-02-01

    A method for visual clone identification of Penicillium commune isolates was developed. The method is based on images of fungal colonies acquired after growth on a standard medium and involves a high degree of objectivity, which in future studies will make it possible for non-experts to perform a qualified identification of different species as well as clones within a species. A total of 77 P. commune isolates from a cheese dairy were 3-point inoculated on Yeast Extract Sucrose (YES) agar and incubated for 7 days at 25 degrees C. After incubation, the isolates were classified into groups containing the same genotype determined by DNA fingerprinting (AFLP). Each genotype also has a specific phenotype such as different colony colours. By careful image acquisition, colours were measured in a reproducible way. Prior to image analysis, each image was corrected with respect to colour, geometry and self-illumination, thereby gaining a set of directly comparable images. A method for automatic extraction of a given number of concentric regions was used. Using the positions of the regions, a number of relevant features--capturing colour and colour-texture from the surface of the fungal colonies--was extracted for further analysis. We introduced the Jeffreys-Matusitas (JM) distance between the feature distributions to express the similarity between regions in two colonies, and to evaluate the overall (weighted) similarity. The nearest neighbour (NN) classification rule was used. On a dataset from 137 isolates, we obtained a "leave-one-out" cross-validation identification rate of approximately 93-98% compared with the result of DNA fingerprinting.

  7. Cell response of Antarctic and temperate strains of Penicillium spp. to different growth temperature.

    PubMed

    Gocheva, Yana G; Krumova, Ekaterina Tz; Slokoska, Lyudmila S; Miteva, Jeny G; Vassilev, Spassen V; Angelova, Maria B

    2006-11-01

    The effect of growth temperature (10, 15, 20, 25, and 30 degrees C) on the cell response was compared between two Antarctic Penicillium sp. strains (Penicillium sp. p14 and Penicillium sp. m12) and a European temperate strain, Penicillium sp. t35. According to the temperature profiles, Penicillium sp. p14 was identified as psychrophilic, while Penicillium sp. m12 and Penicillium sp. t35 as mesophilic fungi, respectively. The results demonstrated that the growth at low temperature does clearly induce oxidative stress events in all strains tested. Decreases in growth temperature below the optimal coincided with markedly enhanced protein carbonyl content, an indicator of oxidatively damaged proteins. Also, the cellular response to growth temperature in terms of reserve carbohydrate was determined. In the mesophilic strains there was essentially no enhancement of glycogen content. This was in contrast to the psychrophilic Penicillium sp. p14, which gradually accumulated glycogen in response to cold (10 degrees C) during the exponential phase. In addition, elevated endogenous levels of trehalose upon low-temperature stress were exhibited by all model microorganisms. Compared with temperate mesophilic Penicillium sp. t35, Antarctic strains (psychrophilic Penicillium sp. p14 and mesophilic Penicillium sp. m12) demonstrated a marked rise in activities of protective enzymes such as superoxide dismutase and catalase at decreasing temperatures. The results suggested that low-temperature resistance is partially associated with enhanced scavenging systems.

  8. Sargassopenillines A–G, 6,6-Spiroketals from the Alga-Derived Fungi Penicillium thomii and Penicillium lividum

    PubMed Central

    Zhuravleva, Olesya I.; Sobolevskaya, Maria P.; Afiyatullov, Shamil Sh.; Kirichuk, Natalya N.; Denisenko, Vladimir A.; Dmitrenok, Pavel S.; Yurchenko, Ekaterina A.; Dyshlovoy, Sergey A.

    2014-01-01

    Seven new 6,6-spiroketals, sargassopenillines A–G (1–7) were isolated from the alga-derived fungi Penicillium thomii KMM 4645 and Penicillium lividum KMM 4663. The structures of these metabolites were determined by HR-MS and 1D and 2D NMR. The absolute configurations of compounds 1, 5 and 6 were assigned by the modified Mosher’s method and by CD data. Sargassopenilline C (3) inhibited the transcriptional activity of the oncogenic nuclear factor AP-1 with an IC50 value of 15 µM. PMID:25501795

  9. Synergistic effect of fungicides on resistant strains of Penicillium italicum and Penicillium digitatum.

    PubMed

    Díaz Borrás, A; Vila Aguilar, R; Hernández Giménez, E

    1988-08-01

    In vitro and in vivo tests were carried out to study the inhibitory effect of mixtures of sodium orthophenylphenate, methyl-1-(butyl-carbamoyl)-2-benzimidazole ('Benomyl') and sodium orthophenylphenate, 2-(4-thiazolil)-benzimidazole ('Thiabendazole') on growth of strains of Penicillium italicum and P. digitatum resistant to sodium orthophenylphenate, 'Benomyl' and 'Thiabendazole'. 'In vitro' tests showed a synergistic effect of the mixtures; 10 ppm of sodium orthophenylphenate and 10 ppm of the benzimidazolic compound in most cases resulted in complete growth inhibition. Navel oranges inoculated with P. italicum spores showed no damage after 7 days of incubation at 25 degrees C and 80-90% relative humidity when treated with a mixture of 100 ppm of sodium orthophenylphenate and 100 ppm of Thiabendazole.

  10. Acidification of apple and orange hosts by Penicillium digitatum and Penicillium expansum.

    PubMed

    Vilanova, L; Viñas, I; Torres, R; Usall, J; Buron-Moles, G; Teixidó, N

    2014-05-16

    New information about virulence mechanisms of Penicillium digitatum and Penicillium expansum could be an important avenue to control fungal diseases. In this study, the ability of P. digitatum and P. expansum to enhance their virulence by locally modulating the pH of oranges and apples was evaluated. For each host, pH changes with a compatible pathogen and a non-host pathogen were recorded, and the levels of different organic acids were evaluated to establish possible relationships with host pH modifications. Moreover, fruits were harvested at three maturity stages to determine whether fruit maturity could affect the pathogens' virulence. The pH of oranges and apples decreased when the compatible pathogens (P. digitatum and P. expansum, respectively) decayed the fruit. The main organic acid detected in P. digitatum-decayed oranges was galacturonic acid produced as a consequence of host maceration in the rot development process. However, the obtained results showed that this acid was not responsible for the pH decrease in decayed orange tissue. The mixture of malic and citric acids could at least contribute to the acidification of P. digitatum-decayed oranges. The pH decrease in P. expansum decayed apples is related to the accumulation of gluconic and fumaric acids. The pH of oranges and apples was not affected when the non-host pathogen was not able to macerate the tissues. However, different organic acid contents were detected in comparison to healthy tissues. The main organic acids detected in P. expansum-oranges were oxalic and gluconic and in P. digitatum-apples were citric, gluconic and galacturonic. Further research is needed to identify the pathogenicity factors of both fungi because the contribution of organic acids has profound implications.

  11. Biotransformation of dehydroepiandrosterone (DHEA) with Penicillium griseopurpureum Smith and Penicillium glabrum (Wehmer) Westling.

    PubMed

    Huang, Li-Hua; Li, Juan; Xu, Gong; Zhang, Xiang-Hua; Wang, Yang-Guang; Yin, Ye-Lin; Liu, Hong-Min

    2010-12-12

    Microbial transformation of dehydroepiandrosterone (DHEA, 1) using Penicillium griseopurpureum Smith and Penicillium glabrum (Wehmer) Westling has been investigated. Neither fungi had been examined previously for steroid biotransformation. One novel metabolic product of DHEA (1) transformed with P. griseopurpureum Smith, 15α-hydroxy-17a-oxa-d-homo-androst-4-ene-3,17-dione (5), was reported for the first time. The steroid products were assigned by interpretation of their spectral data such as (1)H NMR, (13)C NMR, IR, and HR-MS spectroscopy. P. griseopurpureum Smith was proven to be remarkably efficient in oxidation of the DHEA (1) into androst-4-en-3,17-dione (2). The strain was also observed to yield different monooxygenases to introduce hydroxyl groups at C-7α, -14α, and -15α positions of steroids. Preference for Baeyer-Villiger oxidation to lactonize D ring and oxidation of the 3β-alcohol to the 3-ketone were observed in both incubations. The strain of P. glabrum (Wehmer) Westling catalyzed the steroid 1 to generate both testololactone 3, and d-lactone product with 3β-hydroxy-5-en moiety 8. In addition, the strain promoted hydrogenation of the C-5 and C-6 positions, leading to the formation of 3β-hydroxy-17a-oxa-d-homo-5α-androstan-3,17-dione (9). The biotransformation pathways of DHEA (1) with P. glabrum (Wehmer) Westling and P. griseopurpureum Smith have been investigated, respectively. Possible metabolic pathways of DHEA (1) were proposed.

  12. Use of chemosensitization to overcome fludioxonil resistance in Penicillium expansum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Penicillium expansum mutants (FR2 and FR3) resistant to fludioxonil, a phenylpyrrole fungicide, became susceptible through chemosensitization mediated by natural phenolics. Increased sensitivity of FR3 to oxidizing agents, compared to its parental strain (W2), indicated the oxidative stress respons...

  13. Draft Genome Sequence of the Fungus Penicillium solitum NJ1

    PubMed Central

    Zhang, Yuliang; Pennerman, Kayla K.; Hua, Sui Sheng T.; Yu, Jiujiang; Guo, Anping; Liu, Zhixin; Bennett, Joan W.

    2016-01-01

    Penicillium solitum is one of the most prevalent species causing postharvest decay of pomaceous fruits during storage. Here, we report the draft genome of P. solitum strain NJ1, received as a transfer of a strain originally identified as P. griseofulvum by classical means. PMID:27881535

  14. Penicillium menonorum, a new species related to P. pimiteouiense

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Penicillium menonorum is described as a new monoverticillate, non-vesiculate species that resembles P. restrictum and P. pimiteouiense. On the basis of phylogenetic analysis of DNA sequences from four loci, P. menorum occurs in a clade with P. pimiteouiense, P. vinaceum, P. guttulosum, E. rubidurum,...

  15. DOSE-DEPENDENT ALLERGIC ASTHMA RESPONSES TO PENICILLIUM CHRYSOGENUM

    EPA Science Inventory

    ABSTRACT
    Indoor mold has been associated with development of allergic asthma. Penicillium chrysogenum, a common indoor mold, is known to have several allergens and its viable conidia can induce allergic responses in a mouse model of allergic penicilliosis. The hypothesis o...

  16. QUANTITATIVE PCR OF SELECTED ASPERGILLUS, PENICILLIUM AND PAECILOMYCES SPECIES

    EPA Science Inventory

    A total of 65 quantitative PCR (QPCR) assays, incorporating fluorigenic 5' nuclease (TaqMan®) chemistry and directed at the nuclear ribosomal RNA operon, internal transcribed spacer regions (ITS1 or ITS2) was developed and tested for the detection of Aspergillus, Penicillium and ...

  17. EVALUATION OF FUNGAL GROWTH (PENICILLIUM GLABRUM) ON A CEILING TILE

    EPA Science Inventory

    The paper gives results of a study employing static chambers to study the impact of different equilibrium relative humidities (RHs) and moisture conditions on the ability of a new ceiling tile to support fungal growth. Amplification of the mold, Penicillium glabrum, occurred at R...

  18. HEMOLYSIN, CHRYSOLYSIN FROM PENICILLIUM CHRYSOGENUM PROMOTES INFLAMMATORY RESPONSE

    EPA Science Inventory

    Some strains of Penicillium chrysogenum produce a proteinaceous hemolysin, chrysolysin, when incubated on sheep's blood agar at 37 �C but not at 23 �C. Chrysolysin is an aggregating protein composed of approximately 2 kDa monomers, contains one cysteine amino acid, and has an is...

  19. (GTG)5 microsatellite regions in citrinin-producing Penicillium.

    PubMed

    Di Conza, José Alejandro; Nepote, Andrea Fabiana; González, Ana María; Lurá, María Cristina

    2007-03-01

    Morphological and cultural characteristics, as well as biochemical properties, are the main criteria used in fungal taxonomy and in the standard description of fungi species. Sometimes, however, this criterion is difficult to apply due to fungal phenotypic variations. This is particularly true in the genus Penicillium. The aims of this work were to determine (GTG)5 microsatellite sequence in potentially citrinin-producing Penicillium strains and to investigate if this sequence could be useful to characterize such fungi. Penicillium citrinum Thom and Penicillium chrysogenum Thom were isolated from different foods. The identification of the isolates at species level was carried out according to classical taxonomy. The production of citrinin was determined by thin layer chromatography. This study proved that microsatellite regions exist as short repeated sequences in all tested strains. The patterns were very similar for all P. citrinum isolates and it was possible to group them in function of the quantity of citrinin produced. Yet, not similar clusters were obtained when P. chrysogenum isolates were analyzed.

  20. In-hospital source of airborne Penicillium species spores.

    PubMed Central

    Streifel, A J; Stevens, P P; Rhame, F S

    1987-01-01

    Between 16 July and 1 October 1984, prospectively monitored corridor air samples from a bone marrow transplant station revealed a marked increase in airborne thermotolerant Penicillium spores. Simultaneous cultures of outside air showed lower spore counts, which were unchanged before, during, and after the corridor outburst, establishing that the source was within the hospital. Although the corridor was equipped with recirculating high-efficiency particulate air filtration units which provided 16 air changes per h, the mean corridor air count rose to 64.4 thermotolerant Penicillium CFU/m3 during the outburst period. The in-hospital source was ultimately traced to rotting cabinet wood enclosing a sink with leaking pipes in the medication room. It produced approximately 5.5 X 10(5) thermotolerant Penicillium CFU/h. In a patient room supplied by corridor air, an in-room recirculating high-efficiency particulate air filter reduced the mean thermotolerant Penicillium count to 2.2 CFU/m3. No patient illness or colonization occurred as a result of this event, although the cabinet wood, after sterilization, was shown to sustain abundant growth of Aspergillus fumigatus and Aspergillus flavus. Wet organic substrates should be avoided in hospital areas with immunosuppressed patients. Images PMID:3539981

  1. Formation of Diketopiperazines by Penicillium italicum Isolated from Oranges

    PubMed Central

    Scott, Peter M.; Kennedy, Barry P. C.; Harwig, Joost; Chen, Y-K.

    1974-01-01

    Prolyl-2-(1′,1′-dimethylallyl)tryptophyldiketopiperazine and 12,13-dehydroprolyl-2-(1′,1′-dimethylallyl)tryptophyldiketopiperazine, known metabolites of Aspergillus ustus, were produced in low yield by Penicillium italicum in liquid culture and on unsterilized orange peel. PMID:4441068

  2. Antigen detection assay for identification of Penicillium marneffei infection.

    PubMed

    Chaiyaroj, Sansanee C; Chawengkirttikul, Runglawan; Sirisinha, Stitaya; Watkins, Pramuan; Srinoulprasert, Yuttana

    2003-01-01

    Two recently produced monoclonal antibodies were used to develop an antigen capture enzyme-linked immunosorbent assay (ELISA) for rapid diagnosis of Penicillium marneffei. The method was evaluated with 53 patients with culture-confirmed penicilliosis and 240 controls. The diagnostic sensitivity, specificity, and accuracy of the ELISA were 92.45, 97.5, and 96.59%, respectively.

  3. Diversity and enzyme activity of Penicillium species associated with macroalgae in Jeju Island.

    PubMed

    Park, Myung Soo; Lee, Seobihn; Oh, Seung-Yoon; Cho, Ga Youn; Lim, Young Woon

    2016-10-01

    A total of 28 strains of 19 Penicillium species were isolated in a survey of extracellular enzyme-producing fungi from macroalgae along the coast of Jeju Island of Korea. Penicillium species were identified based on morphological and β-tubulin sequence analyses. In addition, the halo-tolerance and enzyme activity of all strains were evaluated. The diversity of Penicillium strains isolated from brown algae was higher than the diversity of strains isolated from green and red algae. The commonly isolated species were Penicillium antarcticum, P. bialowiezense, P. brevicompactum, P. crustosum, P. oxalicum, P. rubens, P. sumatrense, and P. terrigenum. While many strains showed endoglucanase, β-glucosidase, and protease activity, no alginase activity was detected. There was a positive correlation between halo-tolerance and endoglucanase activity within Penicillium species. Among 19 Penicillium species, three species-P. kongii, P. olsonii, and P. viticola-have not been previously recorded in Korea.

  4. Expanding the Species and Chemical Diversity of Penicillium Section Cinnamopurpurea

    PubMed Central

    Peterson, Stephen W.; Jurjević, Željko; Frisvad, Jens C.

    2015-01-01

    A set of isolates very similar to or potentially conspecific with an unidentified Penicillium isolate NRRL 735, was assembled using a BLAST search of ITS similarity among described (GenBank) and undescribed Penicillium isolates in our laboratories. DNA was amplified from six loci of the assembled isolates and sequenced. Two species in section Cinnamopurpurea are self-compatible sexual species, but the asexual species had polymorphic loci suggestive of sexual reproduction and variation in conidium size suggestive of ploidy level differences typical of heterothallism. Accordingly we use genealogical concordance analysis, a technique valid only in heterothallic organisms, for putatively asexual species. Seven new species were revealed in the analysis and are described here. Extrolite analysis showed that two of the new species, P. colei and P. monsserratidens produce the mycotoxin citreoviridin that has demonstrated pharmacological activity against human lung tumors. These isolates could provide leads in pharmaceutical research. PMID:25853891

  5. Isolation and properties of xyloglucanases of Penicillium sp.

    PubMed

    Sinitsyna, O A; Fedorova, E A; Pravilnikov, A G; Rozhkova, A M; Skomarovsky, A A; Matys, V Yu; Bubnova, T M; Okunev, O N; Vinetsky, Yu P; Sinitsyn, A P

    2010-01-01

    Using chromatographic technique, xyloglucanase (XG) A (25 kDa, pI 3.5, 12th glycosyl hydrolase family) was isolated from the enzyme complex secreted by the mycelial fungus Penicillium canescens, and xyloglucanases XG 25 (25 kDa, pI 4.1, 12th glycosyl hydrolase family) and XG 70 (70 kDa, pI 3.5, 74th glycosyl hydrolase family) were isolated from the enzyme complex of Penicillium verruculosum. Properties of the isolated enzymes (substrate specificity, optimal ranges of pH and temperature for enzyme activity and stability, effect of metal ions on catalytic activity) were compared with the properties of xyloglucanases XG 32 of Aspergillus japonicus, XG 78 of Chrysosporium lucknowense, and XG of Trichoderma reesei. The gene xegA encoding XG A of P. canescens was isolated, and the amino acid sequence of the corresponding protein was determined.

  6. Aspergillus and Penicillium identification using DNA sequences: barcode or MLST?

    PubMed

    Peterson, Stephen W

    2012-07-01

    Current methods in DNA technology can detect single nucleotide polymorphisms with measurable accuracy using several different approaches appropriate for different uses. If there are even single nucleotide differences that are invariant markers of the species, we can accomplish identification through rapid DNA-based tests. The question of whether we can reliably detect and identify species of Aspergillus and Penicillium turns mainly upon the completeness of our alpha taxonomy, our species concepts, and how well the available DNA data coincide with the taxonomic diversity in the family Trichocomaceae. No single gene is yet known that is invariant within species and variable between species as would be optimal for the barcode approach. Data are published that would make an MLST approach to isolate identification possible in the most well-studied clades of Aspergillus and Penicillium.

  7. Superactive cellulase formulation using cellobiohydrolase-1 from Penicillium funiculosum

    DOEpatents

    Adney, William S.; Baker, John O.; Decker, Stephen R.; Chou, Yat-Chen; Himmel, Michael E.; Ding, Shi-You

    2012-10-09

    Purified cellobiohydrolase I (glycosyl hydrolase family 7 (Cel7A)) enzymes from Penicillium funiculosum demonstrate a high level of specific performance in comparison to other Cel7 family member enzymes when formulated with purified EIcd endoglucanase from A. cellulolyticus and tested on pretreated corn stover. This result is true of the purified native enzyme, as well as recombinantly expressed enzyme, for example, that enzyme expressed in a non-native Aspergillus host. In a specific example, the specific performance of the formulation using purified recombinant Cel7A from Penicillium funiculosum expressed in A. awamori is increased by more than 200% when compared to a formulation using purified Cel7A from Trichoderma reesei.

  8. Superactive cellulase formulation using cellobiohydrolase-1 from Penicillium funiculosum

    DOEpatents

    Adney, William S.; Baker, John O.; Decker, Stephen R.; Chou, Yat-Chen; Himmel, Michael E.; Ding, Shi-You

    2008-11-11

    Purified cellobiohydrolase I (glycosyl hydrolase family 7 (Cel7A) enzymes from Penicillium funiculosum demonstrate a high level of specific performance in comparison to other Cel7 family member enzymes when formulated with purified EIcd endoglucanase from A. cellulolyticus and tested on pretreated corn stover. This result is true of the purified native enzyme, as well as recombinantly expressed enzyme, for example, that enzyme expressed in a non-native Aspergillus host. In a specific example, the specific performance of the formulation using purified recombinant Cel7A from Penicillium funiculosum expressed in A. awamori is increased by more than 200% when compared to a formulation using purified Cel7A from Trichoderma reesei.

  9. Marine natural products sourced from marine-derived Penicillium fungi.

    PubMed

    Ma, Hong-Guang; Liu, Qiang; Zhu, Guo-Liang; Liu, Hai-Shan; Zhu, Wei-Ming

    2016-01-01

    Marine micro-organisms have been proven to be a major source of marine natural products (MNPs) in recent years, in which filamentous fungi are a vital source of bioactive natural products for their large metagenomes and more complex genetic backgrounds. This review highlights the 390 new MNPs from marine-derived Penicillium fungi during 1991 to 2014. These new MNPs are categorized based on the environment sources of the fungal hosts and their bioactivities are summarized.

  10. Production of mycophenolic acid by Penicillium roqueforti strains.

    PubMed Central

    Lafont, P; Debeaupuis, J P; Gaillardin, M; Payen, J

    1979-01-01

    Sixteen strains of Penicillium roqueforti Thom, isolated from blue-molded cheeses, were studied. In vitro, all of these strains produced mycophenolic acid, some on the order of 0.8 to 4 mg/g od dry culture. The greatest yields were obtained after 10 days of incubation of cultures at 15 degrees C. However, under some experimental conditions, mycophenolic acid was not alone responsible for the toxicity of culture extracts to chicken embryos. PMID:453818

  11. Penicillium subrubescens, a new species efficiently producing inulinase.

    PubMed

    Mansouri, S; Houbraken, J; Samson, R A; Frisvad, J C; Christensen, M; Tuthill, D E; Koutaniemi, S; Hatakka, A; Lankinen, P

    2013-06-01

    Inulin is a reserve carbohydrate in about 15 % of the flowering plants and is accumulated in underground tubers of e.g. chicory, dahlia and Jerusalem artichoke. This carbohydrate consists of linear chains of β-(2,1)-linked fructose attached to a sucrose molecule. Inulinases hydrolyse inulin into fructose and glucose. To find efficient inulin degrading fungi, 126 fungal strains from the Fungal Biotechnology Culture Collection (FBCC) at University of Helsinki and 74 freshly isolated strains from soil around Jerusalem artichoke tubers were screened in liquid cultures with inulin as a sole source of carbon or ground Jerusalem artichoke tubers, which contains up to 19 % (fresh weight) inulin. Inulinase and invertase activities were assayed by the dinitrosalicylic acid (DNS) method and a freshly isolated Penicillium strain originating from agricultural soil (FBCC 1632) was the most efficient inulinase producer. When it was cultivated at pH 6 and 28 °C in 2 litre bioreactors using inulin and Jerusalem artichoke as a carbon source, inulinase and invertase activities were on day 4 7.7 and 3.1 U mL(-1), respectively. The released sugars analysed by TLC and HPLC showed that considerable amounts of fructose were released while the levels of oligofructans were low, indicating an exoinulinase type of activity. Taxonomic study of the inulinase producing strain showed that this isolate represents a new species belonging in Penicillium section Lanata-divaricata. This new species produces a unique combination of extrolites and is phenotypically and phylogenetically closely related to Penicillium pulvillorum. We propose the name Penicillium subrubescens sp. nov. (CBS 132785(T) = FBCC 1632(T)) for this new species.

  12. Role of Penicillic Acid in the Phytotoxicity of Penicillium cyclopium and Penicillium canescens to the Germination of Corn Seeds

    PubMed Central

    Keromnes, Jacqueline; Thouvenot, Daniel

    1985-01-01

    Penicillium cyclopium and Penicillium canescens cultures inhibited the germination of corn. The phytotoxic compound was isolated by solvent extraction and thin-layer chromatography on silica gel. The phytotoxin was identified as penicillic acid by mass spectrometry, nuclear magnetic resonance, and infrared spectroscopy. Gas-liquid chromatography on a capillary glass column separated the two epimeric forms of penicillic acid. The maximum production of penicillic acid was obtained with P. cyclopium cultures grown at 25°C. The phytotoxicity of penicillic acid was manifested by its ability to alter the germination of corn. The percent inhibition of germination was directly proportional to the logarithm of the penicillic acid concentration. Growth of the main root was reduced 50% by concentrations of 500 μg/ml. Images PMID:16346759

  13. A high density COX1 barcode oligonucleotide array for identification and detection of species of Penicillium subgenus Penicillium.

    PubMed

    Chen, W; Seifert, K A; Lévesque, C A

    2009-05-01

    We developed a COX1 barcode oligonucleotide array based on 358 sequences, including 58 known and two new species of Penicillium subgenus Penicillium, and 12 allied species. The array was robotically spotted at near microarray density on membranes. Species and clade-specific oligonucleotides were selected using the computer programs SigOli and Array Designer. Robotic spotting allowed 768 spots with duplicate sets of perfect match and the corresponding mismatch and positive control oligonucleotides, to be printed on 2 × 6 cm(2) nylon membranes. The array was validated with hybridizations between the array and digoxigenin (DIG)-labelled COX1 polymerase chain reaction amplicons from 70 pure DNA samples, and directly from environmental samples (cheese and plants) without culturing. DNA hybridization conditions were optimized, but undesired cross-reactions were detected frequently, reflecting the relatively high sequence similarity of the COX1 gene among Penicillium species. Approximately 60% of the perfect match oligonucleotides were rejected because of low specificity and 76 delivered useful group-specific or species-specific reactions and could be used for detecting certain species of Penicillium in environmental samples. In practice, the presence of weak signals on arrays exposed to amplicons from environmental samples, which could have represented weak detections or weak cross reactions, made interpretation difficult for over half of the oligonucleotides. DNA regions with very few single nucleotide polymorphisms or lacking insertions/deletions among closely related species are not ideal for oligonucleotide-based diagnostics, and supplementing the COX1-based array with oligonucleotides derived from additional genes would result in a more robust hierarchical identification system.

  14. Blue mold to genomics and beyond: Insights into the biology and virulence of phytopathogenic Penicillium species

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pomes, mainly apples and pears, are economically important fruits produced and consumed worldwide. The United States is the second largest producer of pome fruit in the world behind China. Penicillium expansum and other Penicillium spp. are the most common fungal plant pathogens that cause blue mold...

  15. Genome, transcriptome, and functional analyses of Penicillium expansum provide new insights into secondary metabolism and pathogenicity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The relationship between secondary metabolism and infection in pathogenic fungi has remained largely elusive. Penicillium comprises a group of plant pathogens with varying host specificities and with the ability to produce a wide array of secondary metabolites. The genomes of three Penicillium exp...

  16. Genome Sequence of Penicillium solitum RS1, Which Causes Postharvest Apple Decay

    PubMed Central

    Wu, Guangxi; Jurick, Wayne M.; Gaskins, Verneta L.; Yin, Yanbin; Bennett, Joan W.; Shelton, Daniel R.

    2016-01-01

    Penicillium species cause postharvest decay, commonly known as blue mold, in pome fruits, such as apples and pears. To devise novel strategies to prevent and reduce economic losses during storage, the genome sequence of Penicillium solitum RS1 is reported here for the first time. PMID:27174276

  17. Genome Sequencing and Analysis of the Postharvest Fungus Penicillium expansum R21

    PubMed Central

    Zhang, Yuliang; Hua, Sui Sheng T.; Yu, Jiujiang; Bu, Lijing; Pennerman, Kayla K.; Huang, Qixing; Guo, Anping; Bennett, Joan W.

    2017-01-01

    ABSTRACT Blue mold is the vernacular name of a common postharvest disease of stored apples, pears, and quince that is caused by several common species of Penicillium. This study reports the draft genome sequence of Penicillium expansum strain R21, which was isolated from a red delicious apple in 2011 in Pennsylvania. PMID:28209811

  18. Elucidation of the biochemical basis of specificity and pathogenicity of Penicillium digitatum on citrus fruit

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Green mold caused by Penicillium digitatum is the most damaging postharvest diseases of citrus fruit. This Penicillium species is specific to citrus fruit and do not cause progressive decay in any other fresh fruit or vegetable crops. While the etiology of P. digitatum is well understood, the phys...

  19. Genome Sequencing and Analysis of the Postharvest Fungus Penicillium expansum R21.

    PubMed

    Yin, Guohua; Zhang, Yuliang; Hua, Sui Sheng T; Yu, Jiujiang; Bu, Lijing; Pennerman, Kayla K; Huang, Qixing; Guo, Anping; Bennett, Joan W

    2017-02-16

    Blue mold is the vernacular name of a common postharvest disease of stored apples, pears, and quince that is caused by several common species of Penicillium This study reports the draft genome sequence of Penicillium expansum strain R21, which was isolated from a red delicious apple in 2011 in Pennsylvania.

  20. Multi-Locus Analysis of a Citreoviridin-Producing Isolate Previously Identified as Penicillium NRRL 13013

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cole et al (1981) reported a citreoviridin-producing isolate of Penicillium charlesii (NRRL 13013) from molded pecans. Wicklow later identified it as a variant of Penicillium citreoviride, noting that it produced sclerotia, although the species as a whole is not known to do so. We sequenced the IT...

  1. Host ranges of North American isolates of Penicillium causing blue mold of bulb crops

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Single isolates of four Penicillium species belonging to series Corymbifera (Penicillium allii, P. hirsutum, P. tulipae, P. venetum) plus an isolate of P. polonicum, all from North American sources, were inoculated individually into Crocus sativus, Allium sativum (garlic), A. cepa (onion), Iris holl...

  2. Genome sequencing and analyses of the postharvest fungus Penicillium expansum R21

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Blue mold is the vernacular name of a common postharvest disease of stored apples, pears and quince that is caused by several common species of Penicillium. This study reports the draft genome sequence of Penicillium expansum strain R21, a strain isolated from a Red Delicious apple in 2011 in Pennsy...

  3. Clinical, morphological, and molecular characterization of Penicillium canis, sp. nov., isolated from a dog with osteomyelitis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Infections caused by Penicillium spp. are rare in dogs, and the prognosis in these cases is poor. An unknown species of Penicillium was isolated from a bone lesion in a young dog with osteomyelitis of the right ilium. Extensive diagnostic evaluation did not reveal evidence of dissemination. Resoluti...

  4. Augmenting antifungal activity of oxidizing agent with kojic acid: Control of Penicillium strains infecting crops

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Oxidative treatment is a strategy for preventing Penicillium contamination in foods or crops. Antifungal efficacy of oxidant [hydrogen peroxide (H2O2)], biotic effector [kojic acid (KA)] and abiotic stress (heat), alone or in combination, was investigated in Penicillium. The levels of antifungal int...

  5. Genome Sequencing and Analysis of the Filamentous Fungus Penicillium sclerotiorum 113, Isolated after Hurricane Sandy.

    PubMed

    Yin, Guohua; Zhang, Yuliang; Pennerman, Kayla K; Hua, Sui Sheng T; Huang, Qixing; Guo, Anping; Liu, Zhixin; Bennett, Joan W

    2016-11-23

    Penicillium sclerotiorum is a distinctive species within the genus Penicillium that usually produces vivid orange to red colonies, sometimes with colorful sclerotia. Here, we report the first draft genome sequence of P. sclerotiorum strain 113, isolated in 2013 in the aftermath of Hurricane Sandy from a flooded home in New Jersey.

  6. Genome Sequencing and Analysis of the Filamentous Fungus Penicillium sclerotiorum 113, Isolated after Hurricane Sandy

    PubMed Central

    Zhang, Yuliang; Pennerman, Kayla K.; Hua, Sui Sheng T.; Huang, Qixing; Guo, Anping; Liu, Zhixin; Bennett, Joan W.

    2016-01-01

    Penicillium sclerotiorum is a distinctive species within the genus Penicillium that usually produces vivid orange to red colonies, sometimes with colorful sclerotia. Here, we report the first draft genome sequence of P. sclerotiorum strain 113, isolated in 2013 in the aftermath of Hurricane Sandy from a flooded home in New Jersey. PMID:27881534

  7. Genetic and Morphological Diversity of the Genus Penicillium From Mazandaran and Tehran Provinces, Iran

    PubMed Central

    Abastabar, Mahdi; Mirhendi, Hossein; Hedayati, Mohammad Taghi; Shokohi, Tahereh; Rezaei-Matehkolaei, Ali; Mohammadi, Rasoul; Badali, Hamid; Moazeni, Maryam; Haghani, Iman; Ghojoghi, Aynaz; Akhtari, Javad

    2016-01-01

    Background: The genus Penicillium contains a large number of ubiquitous environmental taxa, of which some species are clinically important. Identification of Penicillium down to the species level is currently based on polyphasic criteria, including phenotypic features and genetic markers. Biodiversity of the genus Penicillium from Mazandaran and Tehran provinces has not been described. Objectives: The current paper focused on the environmental biodiversity of Penicillium isolates within some areas of Mazandaran and Tehran provinces, based on morphological traits and the molecular data from partial sequence of the β-tubulin (BT2) gene. Materials and Methods: A total of 400 strains were isolated from the environment and investigated using morphological tests and sequencing of BT2, in order to characterize the spectrum of the Penicillium species. Results: Sequence analysis of BT2 and morphological criteria of 20 strains representative of 10 species showed that Penicillium chrysogenum was the most prevalent species (n = 6), followed by P. polonicum (n = 3), P. glabrum (n = 2), P. palitans (n = 2), P. melanoconidium (n = 2), and other species, including P. expansum, P. canescense, P. griseofulvum, P. italicum, and P. raistrickii with one case each. Conclusions: It was shown that partial β-tubulin sequence, as a reliable genetic target, supported specific morphological criteria for identification of the Penicillium species. Like other assessments throughout the world, P. chrysogenum remains the most frequent environmental Penicillium species in Mazandaran and Tehran Provinces. PMID:27099684

  8. Penicillium brocae, a new species associated with the coffee berry borer in Chiapas, Mexico.

    PubMed

    Peterson, Stephen W; Pérez, Jeanneth; Vega, Fernando E; Infante, Francisco

    2003-01-01

    Penicillium brocae is a new monoverticillate species isolated from coffee berry borers collected at coffee plantations in Mexico near Cacahoatán, Chiapas, and from borers reared on artificial diets at ECOSUR laboratory facilities in Tapachula, Chiapas. Phenotypically, it is in Penicillium series Implicatum, but because it does not conform to known species we have described it as new. ITS and large subunit rDNA were sequenced and compared to determine the phylogenetic position of this species. It is most closely related to Penicillium adametzii. Penicillium brocae has only been found in association with the coffee berry borer and is one of several fungi that grow in coffee berry borer galleries. Penicillium brocae may provide the exogenous sterols necessary for the coffee berry borer's development and thus be mutualistically associated with the insect.

  9. A taxonomic and phylogenetic revision of the Penicillium sclerotiorum complex

    PubMed Central

    Rivera, K.G.; Seifert, K.A.

    2011-01-01

    The morphological concept of Penicillium sclerotiorum (subgenus Aspergilloides) includes strains with monoverticillate, vesiculate conidiophores, and vivid orange to red colony colours, with colourful sclerotia sometimes produced. Multigene phylogenetic analyses with the nuclear ribosomal internal transcribed spacer (ITS) region, cytochrome c oxidase subunit 1 (cox1), β-tubulin (benA), translation elongation factor 1-α (tef1-α), and calmodulin (cmd), reveal that the P. sclerotiorum morphospecies is a complex of seven phylogenetically distinct species, three of which were recently described, namely P. guanacastense, P. mallochii, and P. viticola. Three previously unidentified species are described here as P. cainii, P. jacksonii, and P. johnkrugii. The phylogenetic species are morphologically similar, but differ in combinations of colony characters, sclerotium production, conidiophore stipe roughening and branching, and conidial shape. Ecological characters and differences in geographical distribution further characterise some of the species, but increased sampling is necessary to confirm these differences. The fungal DNA barcode, the ITS, and the animal DNA barcode, cox1, have lower species resolving ability in our phylogenetic analyses, but still allow identification of all the species. Tef1-α and cmd were superior in providing fully resolved, statistically well-supported phylogenetic trees for this species complex, whereas benA resolved all species but had some issues with paraphyly. Penicillium adametzioides and P. multicolor, considered synonyms of P. sclerotiorum by some previous authors, do not belong to the P. sclerotiorum complex. Taxonomic novelties: New species: Penicillium cainii K.G. Rivera, Malloch & Seifert, P. jacksonii K.G. Rivera, Houbraken & Seifert, P. johnkrugii K.G. Rivera, Houbraken & Seifert. PMID:22308047

  10. Dihydroisocoumarins from the Mangrove-Derived Fungus Penicillium citrinum

    PubMed Central

    Huang, Guo-Lei; Zhou, Xue-Ming; Bai, Meng; Liu, Yu-Xin; Zhao, Yan-Lei; Luo, You-Ping; Niu, Yan-Yan; Zheng, Cai-Juan; Chen, Guang-Ying

    2016-01-01

    Three new dihydroisocoumarin penicimarins G–I (1–3), together with one known dihydroisocoumarin (4) and three known meroterpenoids (5–7), were obtained from a fungus Penicillium citrinum isolated from the mangrove Bruguiera sexangula var. rhynchopetala collected in the South China Sea. Their structures were elucidated by the detailed analysis of spectroscopic data. The absolute configuration of 1 was determined by the X-ray diffraction analysis using Cu Kα radiation. The absolute configurations of 2 and 3 were determined by comparison of their circular dichroism (CD) spectra with the literature. All compounds were evaluated for their antibacterial activities and cytotoxic activities. PMID:27735855

  11. An antifungal tetrapeptide from the culture of Penicillium canescens.

    PubMed

    Bertinetti, Brenda V; Peña, Nora I; Cabrera, Gabriela M

    2009-08-01

    A new tetrapeptide D-Phe-L-Val-D-Val-L-Tyr (1), along with three known diketopiperazines and pseurotin A, were isolated from the culture of Penicillium canescens, collected from pollen from beehives, in a screening for new antimicrobial products from unexplored sources. The structure of the tetrapeptide, which exhibits antifungal activity comparable with that of the commercial product benomyl against the soybean phytopathogen Fusarium virguliforme, was determined by spectroscopic (2D-NMR, and MS and MS/MS) and chemical methods, and the sequence was confirmed by comparison with authentic synthetic isomeric peptides.

  12. [Biological activity of Penicillium sp. 10-51 exometabolites].

    PubMed

    Savchuk, Ia I; Zaĭchenko, A M; Tsyganenko, E S

    2012-01-01

    Silica gel column chromatography (silica gel "L" II kind of activity 100/160 mkm) of the chloroform extract from the cultural filtrate of Penicillium sp. 10-51 gave two fractions (chloroform and chloroform-acetone, 5:1) having biological activity. Recrystallization yielded two compounds. On the basis of physico-chemical and spectral data these compounds were identified as curvularin and hydroxycurvularin, which have a large spectrum of biological action as to bacteria, yeast, blue-green algae and phytopathogenic micromycetes.

  13. Cyclohexanone derivatives with cytotoxicity from the fungus Penicillium commune.

    PubMed

    Liu, Fang-zhi; Ren, Jin-wei; Tang, Jin-shan; Liu, Xing-zhong; Che, Yong-sheng; Yao, Xin-sheng

    2013-06-01

    Four new cyclohexanone derivatives (2-5) and one known analog, (-)-Palitantin (1) were isolated from the EtOAc extract of Penicillium commune, a fungal strain of low-temperature habitats. The structures of 2-5 were determined on the basis of extensive spectroscopic analysis. Furthermore, the absolute configuration of 2 was assigned by electronic circular dichroism (ECD) calculations, whereas that 3-5 were deduced via the CD data. Cytotoxicities of 2-5 against five human carcinoma cell lines (Hela, A549, MCF7, HCT116, T24) were evaluated.

  14. Penicillium digitatum metabolites on synthetic media and citrus fruits.

    PubMed

    Ariza, Marta R; Larsen, Thomas O; Peterson, Bent O; Duus, Jens O; Barrero, Alejandro F

    2002-10-23

    Penicillium digitatum has been cultured on citrus fruits and yeast extract sucrose agar media (YES). Cultivation of fungal cultures on solid medium allowed the isolation of two novel tryptoquivaline-like metabolites, tryptoquialanine A (1) and tryptoquialanine B (2), also biosynthesized on citrus fruits. Their structural elucidation is described on the basis of their spectroscopic data, including those from 2D NMR experiments. The analysis of the biomass sterols led to the identification of 8-12. Fungal infection on the natural substrates induced the release of citrus monoterpenes together with fungal volatiles. The host-pathogen interaction in nature and the possible biological role of citrus volatiles are also discussed.

  15. Experimental and theoretical investigations of the adhesion time of Penicillium spores to cedar wood surface.

    PubMed

    Soumya, Elabed; Ibnsouda, Saad Koraichi; Abdellah, Houari; Hassan, Latrache

    2013-04-01

    In this study, the adhesion of 4 Penicillium strains (Penicillium granulatum, Penicillium crustosum, Penicillium commune and Penicillium chrysogenum) on cedar wood was examined qualitatively and quantitatively by using the extended DLVO (XDLVO) approach and the environmental scanning electronic microscopy (ESEM) technique. A comparison between the XDLVO theories and the ESEM technique was also investigated. The adhesion tests revealed that P. chrysogenum was not able to adhere on the cedar wood substrata, as predicted by the XDLVO approach. We have also found by ESEM that the three Penicillium strains (P. granulatum, P. crustosum, P. commune) adhered on wood, as not predicted theoretically. Moreover, the time of adhesion (3 h and 24 h) was used not only to compare the capacity of adhesion according to contact time but also to explain the discrepancies between the XDLVO approach prediction and the adhesion experiments. A positive relationship between the XDLVO approach and adhesion experiments has been observed after 3h of adhesion. In contrast, a contradiction between the XDLVO predictions and the adhesion test results has been noted after 24h of adhesion of Penicillium strains to the wood surface.

  16. Penicillium strains isolated from Slovak grape berries taxonomy assessment by secondary metabolite profile.

    PubMed

    Santini, Antonello; Mikušová, Petra; Sulyok, Michael; Krska, Rudolf; Labuda, Roman; Srobárová, Antónia

    2014-11-01

    The secondary metabolite profiles of microfungi of the genus Penicillium isolated from samples of grape berries collected in two different phases during two vegetative seasons in Slovakia is described to assess the taxonomy. Three Slovak vine regions have been selected for this study, based on their climatic differences and national economic importance. Cultures of microfungi isolated from berries were incubated on different selective media for macro and micromorphology identification. The species Penicillium brevicompactum, Penicillium crustosum, Penicillium chrysogenum, Penicillium expansum, Penicillium palitans and Penicillium polonicum were identified according to growth and morphology. The related strains were found to produce a broad spectrum of fungal metabolites, including roquefortine C, chaetoglobosin A, penitrem A, cyclopeptin, cyclopenin, viridicatin, methylviridicatin, verrucofortine, secalonic acid D, cyclopiazonic acid, fumigaclavine and mycophenolic acid. Chemotaxonomy was performed using high-performance liquid chromatography (HPLC) and mass spectrometry (MS). Dried grape berries were also analyzed allowing to assess the presence of patulin, roquefortine C and penicillic acid; this last one has been identified in dried berries but not in vitro.

  17. New promoters for strain engineering of Penicillium chrysogenum.

    PubMed

    Polli, Fabiola; Meijrink, Ben; Bovenberg, Roel A L; Driessen, Arnold J M

    2016-04-01

    Filamentous fungi such as Aspergillus and Penicillium are widely used as hosts for the industrial products such as proteins and secondary metabolites. Although filamentous fungi are versatile in recognizing transcriptional and translational elements present in genes from other filamentous fungal species, only few promoters have been applied and compared in performance so far in Penicillium chrysogenum. Therefore, a set of homologous and heterologous promoters were tested in a reporter system to obtain a set of potential different strengths. Through in vivo homologous recombination in Saccharomyces cerevisiae, twelve Aspergillus niger and P. chrysogenum promoter-reporter pathways were constructed that drive the expression of green fluorescent protein while concurrent expression of the red fluorescent protein was used as an internal standard and placed under control of the PcPAF promoter. The pathways were integrated into the genome of P. chrysogenum and tested using the BioLector system for fermentation. Reporter gene expression was monitored during growth and classified according to promoter strength and expression profile. A set of novel promoters was obtained that can be used to tune the expression of target genes in future strain engineering programs.

  18. Massive gene swamping among cheese-making Penicillium fungi

    PubMed Central

    Ropars, Jeanne; Aguileta, Gabriela; de Vienne, Damien M.; Giraud, Tatiana

    2014-01-01

    Horizontal gene transfers (HGT), i.e., the transmission of genetic material between species not directly attributable to meiotic gene exchange, have long been acknowledged as a major driver of prokaryotic evolution and is increasingly recognized as an important source of adaptation in eukaryotes. In fungi in particular, many convincing examples of HGT have been reported to confer selective advantages on the recipient fungal host, either promoting fungal pathogenicity on plants or increasing their toxicity by the acquisition of secondary metabolic clusters, resulting in adaptation to new niches and in some cases eventually even in speciation. These horizontal gene transfers involve single genes, complete metabolic pathways or even entire chromosomes. A recent study has uncovered multiple recent horizontal transfers of a 575 kb genomic island in cheese Penicillium fungi, representing ca. 2% of the Penicillium roqueforti’s genome, that may confer selective advantage in the competing cheese environment where bacteria and fungi occur. Novel phylogenomic methods are being developed, revealing massive HGT among fungi. Altogether, these recent studies indicate that HGT is a crucial mechanism of rapid adaptation, even among eukaryotes.

  19. Detection of Penicillium expansum by polymerase chain reaction.

    PubMed

    Marek, Patrick; Annamalai, Thirunavukkarasu; Venkitanarayanan, Kumar

    2003-12-31

    Penicillium expansum is a major causative agent of postharvest decay in a variety of fruits, including apples, peaches, nectarines, and cherries. It causes significant economic losses to the fruit industry and is also of potential public health significance, since it produces patulin, a mycotoxin known to cause harmful effects in animals. Rapid and specific detection of P. expansum is important for ensuring microbiological quality and safety of fruits and fruit juices. The traditional methods for identification of P. expansum are time-consuming and labor-intensive. In this study, we report a polymerase chain reaction utilizing primers based on the polygalacturonase gene of P. expansum. The PCR amplified a 404-bp DNA product from all the P. expansum isolates tested, but not in other common foodborne Penicillium species and Escherichia coli. Experiments to determine the sensitivity of the PCR indicated that it can detect the DNA equivalent from as low as 25 spores of P. expansum. The PCR could potentially be used as a rapid tool for screening fruits for the presence of P. expansum.

  20. Penicillium kongii, a new terverticillate species isolated from plant leaves in China.

    PubMed

    Wang, Bo; Wang, Long

    2013-01-01

    A new Penicillium species isolated from plant leaves, characterized by restricted growth, terverticillate penicilli, ovoid to ellipsoidal conidia and a red soluble pigment on yeast extract sucrose agar is reported here. Penicillium kongii sp. nov. belongs to subgenus Penicillium section Brevicompacta and is morphologically similar to P. bialowiezense and P. brevicompactum. Phylogenetic analyses based on sequence data from calmodulin gene, β-tubulin gene and rDNA ITS1-5.8S-ITS2 show that P. kongii forms a distinctive clade.

  1. Production of penicillin by fungi growing on food products: identification of a complete penicillin gene cluster in Penicillium griseofulvum and a truncated cluster in Penicillium verrucosum.

    PubMed

    Laich, Federico; Fierro, Francisco; Martín, Juan F

    2002-03-01

    Mycobiota growing on food is often beneficial for the ripening and development of the specific flavor characteristics of the product, but it can also be harmful due to the production of undesirable compounds such as mycotoxins or antibiotics. Some of the fungi most frequently isolated from fermented and cured meat products such as Penicillium chrysogenum and Penicillium nalgiovense are known penicillin producers; the latter has been shown to be able to produce penicillin when growing on the surface of meat products and secrete it to the medium. The presence of penicillin in food must be avoided, since it can lead to allergic reactions and the arising of penicillin resistance in human-pathogenic bacteria. In this article we describe a study of the penicillin production ability among fungi of the genus Penicillium that are used as starters for cheese and meat products or that are frequently isolated from food products. Penicillium griseofulvum was found to be a new penicillin producer and to have a penicillin gene cluster similar to that of Penicillium chrysogenum. No other species among the studied fungi were found to produce penicillin or to possess the penicillin biosynthetic genes, except P. verrucosum, which contains the pcbAB gene (as shown by hybridization and PCR cloning of fragments of the gene) but lacks pcbC and penDE. Antibacterial activities due to the production of secondary metabolites other than penicillin were observed in some fungi.

  2. Post-harvest proteomics of grapes infected by Penicillium during withering to produce Amarone wine.

    PubMed

    Lorenzini, Marilinda; Mainente, Federica; Zapparoli, Giacomo; Cecconi, Daniela; Simonato, Barbara

    2016-05-15

    The study of withered grape infection by Penicillium, a potentially toxigenic fungus, is relevant to preserve grape quality during the post-harvest dehydration process. This report describes the first proteomic analysis of Amarone wine grapes, infected by two strains of Penicillium expansum (Pe1) and Penicillium crustosum (Pc4). Protein identification by MS analysis allowed a better understanding of physiological mechanisms underlying the pathogen attack. The Pe1 strain had a major impact on Vitis vinifera protein expression inducing pathogenesis-related proteins and other protein species involved in energy metabolism. A greater expression of new Penicillium proteins involved in energy metabolism and some protein species related to redox homeostasis has been observed on grapes infected by Pc4 strain. Moreover, the new induced proteins in infected grapes could represent potential markers in withered grapes, thus creating the chance to develop case-sensitive prevention strategies to inhibit fungal growth.

  3. Phylogeny and nomenclature of the genus Talaromyces and taxa accommodated in Penicillium subgenus Biverticillium

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The taxonomic history of Penicillium subgenus Biverticillium is reviewed, along with evidence supporting its relationship to Talaromyces. The phylogenetic relationships of these two groups with other genera of Trichocomaceae was studied by sequencing the internal transcribed spacer (ITS) region and ...

  4. Molecular identification of species from the Penicillium roqueforti group associated with spoiled animal feed.

    PubMed

    Boysen, M E; Jacobsson, K G; Schnürer, J

    2000-04-01

    The Penicillium roqueforti group has recently been split into three species, P. roqueforti, Penicillium carneum, and Penicillium paneum, on the basis of differences in ribosomal DNA sequences and secondary metabolite profiles. We reevaluated the taxonomic identity of 52 livestock feed isolates from Sweden, previously identified by morphology as P. roqueforti, by comparing the sequences of the ribosomal internal transcribed spacer region. Identities were confirmed with random amplified polymorphic DNA analysis and secondary metabolite profiles. Of these isolates, 48 were P. roqueforti, 2 were P. paneum, and 2 were Penicillium expansum. No P. carneum isolates were found. The three species produce different mycotoxins, but no obvious relationship between mold and animal disease was detected, based on medical records. P. roqueforti appears to dominate in silage, but the ecological and toxicological importance of P. carneum and P. paneum as feed spoilage fungi is not clear. This is the first report of P. expansum in silage.

  5. Compartmentalization in penicillin G biosynthesis by Penicillium chrysogenum PQ-96.

    PubMed

    Kurzątkowski, Wiesław; Staniszewska, Monika; Bondaryk, Małgorzata; Gębska-Kuczerowska, Anita

    2014-01-01

    The arrangement of organelles in the sub-apical productive non-growing vacuolated hyphal cells of the high- and the low-penicillin-pro- ducing strains Penicillium chrysogenum was compared using transmission electron microscopy. In the productive cells of the high-yielding strain the endoplasmic reticulum and the polyribosomes with associated peroxisomes are frequently arranged at the periphery of the cytoplasm and around the vacuoles. At the high activity of penicillin G biosynthesis the immuno-label of the cytosolic isopenicillin N synthase is concentrated at the polyribosomes arranged in the peripheral cytoplasm and along the tonoplast as well as around the peroxisomes. On the basis of the obtained results the compartmentalization of the pathway of penicillin G biosymthesis is discussed. The obtained results support the phenylacetic acid detoxification hypothesis of penicillin G biosynthesis.

  6. Autophagy deficiency promotes beta-lactam production in Penicillium chrysogenum.

    PubMed

    Bartoszewska, Magdalena; Kiel, Jan A K W; Bovenberg, Roel A L; Veenhuis, Marten; van der Klei, Ida J

    2011-02-01

    We have investigated the significance of autophagy in the production of the β-lactam antibiotic penicillin (PEN) by the filamentous fungus Penicillium chrysogenum. In this fungus PEN production is compartmentalized in the cytosol and in peroxisomes. We demonstrate that under PEN-producing conditions significant amounts of cytosolic and peroxisomal proteins are degraded via autophagy. Morphological analysis, based on electron and fluorescence microscopy, revealed that this phenomenon might contribute to progressive deterioration of late subapical cells. We show that deletion of the P. chrysogenum ortholog of Saccharomyces cerevisiae serine-threonine kinase atg1 results in impairment of autophagy. In P. chrysogenum atg1 cells, a distinct delay in cell degeneration is observed relative to wild-type cells. This phenomenon is associated with an increase in the enzyme levels of the PEN biosynthetic pathway and enhanced production levels of this antibacterial compound.

  7. Improving penicillin biosynthesis in Penicillium chrysogenum by glyoxalase overproduction.

    PubMed

    Scheckhuber, Christian Q; Veenhuis, Marten; van der Klei, Ida J

    2013-07-01

    Genetic engineering of fungal cell factories mainly focuses on manipulating enzymes of the product pathway or primary metabolism. However, despite the use of strong promoters or strains containing the genes of interest in multiple copies, the desired strongly enhanced enzyme levels are often not obtained. Here we present a novel strategy to improve penicillin biosynthesis by Penicillium chrysogenum by reducing reactive and toxic metabolic by-products, 2-oxoaldehydes. This was achieved by overexpressing the genes encoding glyoxalase I and II, which resulted in a 10% increase in penicillin titers relative to the control strain. The protein levels of two key enzymes of penicillin biosynthesis, isopenicillin N synthase and isopenicillin N acyltransferase, were increased in the glyoxalase transformants, whereas their transcript levels remained unaltered. These results suggest that directed intracellular reduction of 2-oxoaldehydes prolongs the functional lifetime of these enzymes.

  8. The paf gene product modulates asexual development in Penicillium chrysogenum.

    PubMed

    Hegedüs, Nikoletta; Sigl, Claudia; Zadra, Ivo; Pócsi, Istvan; Marx, Florentine

    2011-06-01

    Penicillium chrysogenum secretes a low molecular weight, cationic and cysteine-rich protein (PAF). It has growth inhibitory activity against the model organism Aspergillus nidulans and numerous zoo- and phytopathogenic fungi but shows only minimal conditional antifungal activity against the producing organism itself. In this study we provide evidence for an additional function of PAF which is distinct from the antifungal activity against putative ecologically concurrent microorganisms. Our data indicate that PAF enhances conidiation in P. chrysogenum by modulating the expression of brlA, the central regulatory gene for mitospore development. A paf deletion strain showed a significant impairment of mitospore formation which sustains our hypothesis that PAF plays an important role in balancing asexual differentiation in P. chrysogenum.

  9. Pectin Lyase Production by a Penicillium italicum Strain

    PubMed Central

    Alaña, Aitor; Gabilondo, Ane; Hernando, Fernando; Moragues, Maria D.; Dominguez, Juan B.; Llama, Maria J.; Serra, Juan L.

    1989-01-01

    Growth and concomitant production of an extracellular pectin lyase (PL) [poly(methoxylgalactosiduronate) endolyase; EC 4.2.2.10] were investigated in a group of 16 fungi grown in liquid medium containing pectin as a supplementary carbon source. Culture filtrates of both Penicillium italicum (CECT 2294) and P. expansum (CECT 2275) showed the highest PL activity and contained polygalacturonase but not pectinesterase activity. The effect of the inoculum size, the carbon source (sucrose and glucose syrup), and the presence of pectin on the production of PL by P. italicum was studied. The presence of 2.6 mM glycerophosphate in the culture medium enhanced the appearance of PL but was not inhibitory for the in vitro activity. However, glycerol inhibited the enzyme nearly 50% at such a concentration. PMID:16347954

  10. Effects of carbon dioxide on Penicillium chrysogenum: an autoradiographic study

    SciTech Connect

    Edwards, A.G.; Ho, C.S.

    1988-06-20

    Previous research has shown that dissolved carbon dioxide causes significant changes in submerged penicillin fermentations, such as stunted, swollen hyphae, increased branching, lower growth rates, and lower penicillin productivity. Influent carbon dioxide levels of 5 and 10% were shown through the use of autoradiography to cause an increase in chitin synthesis in submerged cultures of Penicillium chrysogenum. At an influent 5% carbon dioxide level, chitin synthesis is ca. 100% greater in the subapical region of P. chrysogenum hyphae than that of the control, in which there was no influent carbon dioxide. Influent carbon dioxide of 10% caused an increase of 200% in chitin synthesis. It is believed that the cell wall must be plasticized before branching can occur and that high amounts of dissolved carbon dioxide cause the cell to lose control of the plasticizing effect, thus the severe morphological changes occur.

  11. Heterologous Expression of Two Ferulic Acid Esterases from Penicillium funiculosum

    NASA Astrophysics Data System (ADS)

    Knoshaug, Eric P.; Selig, Michael J.; Baker, John O.; Decker, Stephen R.; Himmel, Michael E.; Adney, William S.

    Two recombinant ferulic acid esterases from Penicillium funiculosum produced in Aspergillus awamori were evaluated for their ability to improve the digestibility of pretreated corn stover. The genes, faeA and faeB, were cloned from P. funiculosum and expressed in A. awamori using their native signal sequences. Both enzymes contain a catalytic domain connected to a family 1 carbohydrate-binding module by a threonine-rich linker peptide. Interestingly, the carbohydrate binding-module is N-terminal in FaeA and C-terminal in FaeB. The enzymes were purified to homogeneity using column chromatography, and their thermal stability was characterized by differential scanning microcalorimetry. We evaluated both enzymes for their potential to enhance the cellulolytic activity of purified Trichoderma reesei Cel7A on pretreated corn stover.

  12. Hypersensitivity pneumonitis induced by Penicillium expansum in a home environment.

    PubMed

    Park, H S; Jung, K S; Kim, S O; Kim, S J

    1994-04-01

    An episode of fever, cough, shortness of breath and leucocytosis developed in a 31-year-old atopic housewife from mould exposure in her home environment is evaluated. A chest radiograph revealed diffuse tiny nodular infiltrations in both whole lung fields. Spirometry revealed a severe restrictive type of ventilation impairment. Bronchoalveolar lavage (BAL) showed an increased lymphocyte count with reversed CD4+/CD8+ ratio and transbronchial lung biopsy showed markedly increased lymphocytic infiltration in alveolar septa. Fungal cultures in the air of her home were positive for Penicillium expansum and other fungi. Double immunodiffusion test with the patient's serum showed two precipitin bands to P. expansum antigens. Her symptoms, abnormal findings of radiograph, and spirometric abnormalities disappeared after 2 months' avoidance. The serum precipitin disappeared after 1 month's avoidance. This study indicates that the patient had hypersensitivity pneumonitis (HP) on exposure to P. expansum in her home environment.

  13. Heterologous Expression of Two Ferulic Acid Esterases from Penicillium Funiculosum

    SciTech Connect

    Knoshaug, E. P.; Selig, M. J.; Baker, J. O.; Decker, S. R.; Himmel, M. E.; Adney, W. S.

    2008-01-01

    Two recombinant ferulic acid esterases from Penicillium funiculosum produced in Aspergillus awamori were evaluated for their ability to improve the digestibility of pretreated corn stover. The genes, faeA and faeB, were cloned from P. funiculosum and expressed in A. awamori using their native signal sequences. Both enzymes contain a catalytic domain connected to a family 1 carbohydrate-binding module by a threonine-rich linker peptide. Interestingly, the carbohydrate binding-module is N-terminal in FaeA and C-terminal in FaeB. The enzymes were purified to homogeneity using column chromatography, and their thermal stability was characterized by differential scanning microcalorimetry. We evaluated both enzymes for their potential to enhance the cellulolytic activity of purified Trichoderma reesei Cel7A on pretreated corn stover.

  14. Removal of lead from aqueous solutions by Penicillium biomass

    SciTech Connect

    Hui Niu; Xue Shu Xu; Jian Hua Wang . Dept. of Chemical Engineering); Volesky, B. . Dept. of Chemical Engineering)

    1993-09-05

    The removal of lead ions from aqueous solutions by adsorption on nonliving Penicillium chrysogenum biomass was studied. Biosorption of the Pb[sup +2] ion was strongly affected by pH. Within a pH range of 4 to 5, the saturated sorption uptake of Pb[sup +2] was 116 mg/g dry biomass, higher than that of activated charcoal and some other microorganisms. At pH 4.5, P. chrysogenum biomass exhibited selectivity for Pb[sup +2] over other metal ions such as Cd[sup +2], Cu[sup +2], Zn [sup +2], and As[sub +3]. Sorption preference for metals decreased in the following order: Pb > Cd > Cu > Zn > As. The sorption uptake of Pb[sup +2] remained unchanged in the presence of Cu[sup +2] and As [sup +3], it decreased in the presence of Zn[sup +2], and increased in the presence of Cd[sup +2].

  15. Oxidation mechanism of Penicillium digitatum spores through neutral oxygen radicals

    NASA Astrophysics Data System (ADS)

    Hashizume, Hiroshi; Ohta, Takayuki; Takeda, Keigo; Ishikawa, Kenji; Hori, Masaru; Ito, Masafumi

    2014-01-01

    To investigate the inactivation process of Penicillium digitatum spores through neutral oxygen species, the spores were treated with an atmospheric-pressure oxygen radical source and observed in-situ using a fluorescent confocal-laser microscope. The treated spores were stained with two fluorescent dyes, 1,1‧-dioctadecyl-3,3,Y,3‧-tetramethylindocarbocyanine perchlorate (DiI) and diphenyl-1-pyrenylphosphine (DPPP). The intracellular organelles as well as the cell membranes in the spores treated with the oxygen radical source were stained with DiI without a major morphological change of the membranes. DPPP staining revealed that the organelles were oxidized by the oxygen radical treatment. These results suggest that neutral oxygen species, especially atomic oxygen, induce a minor structural change or functional inhibition of cell membranes, which leads to the oxidation of the intracellular organelles through the penetration of reactive oxygen species into the cell.

  16. Molecular identification of Penicillium marneffei using rolling circle amplification.

    PubMed

    Sun, Jiufeng; Najafzadeh, M J; Zhang, Junmin; Vicente, V A; Xi, Liyan; de Hoog, G S

    2011-11-01

    Penicillium marneffei is the aetiological agent of a severe systemic disease in immunocompromised hosts in Southeast Asia. In the present study, we evaluated an identification method based on rolling circle amplification (RCA) enabling rapid and specific detection of single nucleotide differences. Three padlock probes were designed on the basis of the internal transcribed spacers 1 and 2 (ITS) of the rRNA operon. One of these (PmPL1) allowed specific amplification of P. marneffei DNA within one working day using a newly conceived protocol, while no cross-reactivity was observed with other fungi including related biverticillate penicillia. Amplification products can be detected by electrophoresis on agarose gel. The method provides a powerful tool for a rapid specific identification of P. marneffei in the clinical laboratory and has potential for ecological studies.

  17. Isolation and characterization of a novel mycovirus from Penicillium digitatum

    SciTech Connect

    Niu, Yuhui; Zhang, Tingfu; Zhu, Ying; Yuan, Yongze; Wang, Shengqiang; Liu, Jing; Liu, Deli

    2016-07-15

    A novel double-stranded RNA virus designated Penicillium digitatum virus 1 (PdV1) was isolated from the citrus fruit rot pathogen P. digitatum (HS-RH1). The full-length cDNA sequence of the dsRNA/PdV1 (5211 bp) possesses two partially overlapping open reading frames, which encode a coat protein (CP) and a putative RNA-dependent RNA polymerase (RdRp), respectively. Phylogenetic analysis based on multiple alignments of the amino acid sequences of the RdRp and CP indicated that PdV1 tentatively belongs to the genus Victorivirus in the Totiviridae family. Electron micrographs of negatively stained viral particles purified from the peak fraction of sucrose density gradient centrifugation showed spherical particles ~35 nm in diameter. Transfection experiments with purified virions indicated that PdV1 could reduce the vegetative growth and virulence of P. digitatum strain HS-F6. In summary, we report the first isolation and characterization of a mycovirus from P. digitatum that contributes to the hypovirulence phenotypes of the host strain. - Highlights: • A novel victorivirus designated Penicillium digitatum virus 1 (PdV1) was isolated fromP. digitatum. • The dsRNA genome of PdV1 are 5211 bp long, two ORFs encoding CP and RdRp, and are encased in virions of ~35 nm in diameter. • PdV1 infection led to hypovirulent effect on P. digitatum. PdV1 may potentially be used for citrus green mold biocontrol. • Our study provides a research basis for establishing a model system for the study of P. digitatum–mycovirus interactions.

  18. High throughput de novo RNA sequencing elucidates novel responses in Penicillium chrysogenum under microgravity.

    PubMed

    Sathishkumar, Yesupatham; Krishnaraj, Chandran; Rajagopal, Kalyanaraman; Sen, Dwaipayan; Lee, Yang Soo

    2016-02-01

    In this study, the transcriptional alterations in Penicillium chrysogenum under simulated microgravity conditions were analyzed for the first time using an RNA-Seq method. The increasing plethora of eukaryotic microbial flora inside the spaceship demands the basic understanding of fungal biology in the absence of gravity vector. Penicillium species are second most dominant fungal contaminant in International Space Station. Penicillium chrysogenum an industrially important organism also has the potential to emerge as an opportunistic pathogen for the astronauts during the long-term space missions. But till date, the cellular mechanisms underlying the survival and adaptation of Penicillium chrysogenum to microgravity conditions are not clearly elucidated. A reference genome for Penicillium chrysogenum is not yet available in the NCBI database. Hence, we performed comparative de novo transcriptome analysis of Penicillium chrysogenum grown under microgravity versus normal gravity. In addition, the changes due to microgravity are documented at the molecular level. Increased response to the environmental stimulus, changes in the cell wall component ABC transporter/MFS transporters are noteworthy. Interestingly, sustained increase in the expression of Acyl-coenzyme A: isopenicillin N acyltransferase (Acyltransferase) under microgravity revealed the significance of gravity in the penicillin production which could be exploited industrially.

  19. Clinical, Morphological, and Molecular Characterization of Penicillium canis sp. nov., Isolated from a Dog with Osteomyelitis

    PubMed Central

    Sutton, Deanna A.; Swenson, Cheryl L.; Bailey, Chris J.; Wiederhold, Nathan P.; Nelson, Nathan C.; Thompson, Elizabeth H.; Wickes, Brian L.; French, Stephanie; Fu, Jianmin; Vilar-Saavedra, Paulo

    2014-01-01

    Infections caused by Penicillium species are rare in dogs, and the prognosis in these cases is poor. An unknown species of Penicillium was isolated from a bone lesion in a young dog with osteomyelitis of the right ilium. Extensive diagnostic evaluation did not reveal evidence of dissemination. Resolution of lameness and clinical stability of disease were achieved with intravenous phospholipid-complexed amphotericin B initially, followed by long-term combination therapy with terbinafine and ketoconazole. A detailed morphological and molecular characterization of the mold was undertaken. Sequence analysis of the internal transcribed spacer revealed the isolate to be closely related to Penicillium menonorum and Penicillium pimiteouiense. Additional sequence analysis of β-tubulin, calmodulin, minichromosome maintenance factor, DNA-dependent RNA polymerase, and pre-rRNA processing protein revealed the isolate to be a novel species; the name Penicillium canis sp. nov. is proposed. Morphologically, smooth, ovoid conidia, a greenish gray colony color, slow growth on all media, and a failure to form ascomata distinguish this species from closely related Penicillium species. PMID:24789186

  20. Clinical, morphological, and molecular characterization of Penicillium canis sp. nov., isolated from a dog with osteomyelitis.

    PubMed

    Langlois, Daniel K; Sutton, Deanna A; Swenson, Cheryl L; Bailey, Chris J; Wiederhold, Nathan P; Nelson, Nathan C; Thompson, Elizabeth H; Wickes, Brian L; French, Stephanie; Fu, Jianmin; Vilar-Saavedra, Paulo; Peterson, Stephen W

    2014-07-01

    Infections caused by Penicillium species are rare in dogs, and the prognosis in these cases is poor. An unknown species of Penicillium was isolated from a bone lesion in a young dog with osteomyelitis of the right ilium. Extensive diagnostic evaluation did not reveal evidence of dissemination. Resolution of lameness and clinical stability of disease were achieved with intravenous phospholipid-complexed amphotericin B initially, followed by long-term combination therapy with terbinafine and ketoconazole. A detailed morphological and molecular characterization of the mold was undertaken. Sequence analysis of the internal transcribed spacer revealed the isolate to be closely related to Penicillium menonorum and Penicillium pimiteouiense. Additional sequence analysis of β-tubulin, calmodulin, minichromosome maintenance factor, DNA-dependent RNA polymerase, and pre-rRNA processing protein revealed the isolate to be a novel species; the name Penicillium canis sp. nov. is proposed. Morphologically, smooth, ovoid conidia, a greenish gray colony color, slow growth on all media, and a failure to form ascomata distinguish this species from closely related Penicillium species.

  1. Marine-derived Penicillium in Korea: diversity, enzyme activity, and antifungal properties.

    PubMed

    Park, Myung Soo; Fong, Jonathan J; Oh, Seung-Yoon; Kwon, Kae Kyoung; Sohn, Jae Hak; Lim, Young Woon

    2014-08-01

    The diversity of marine-derived Penicillium from Korea was investigated using morphological and multigene phylogenetic approaches, analyzing sequences of the internal transcribed spacer region, β-tubulin gene, and RNA polymerase subunit II gene. In addition, the biological activity of all isolated strains was evaluated. We tested for the extracellular enzyme activity of alginase, endoglucanase, and β-glucosidase, and antifungal activity against two plant pathogens (Colletotrichum acutatum and Fusarium oxysporum). A total of 184 strains of 36 Penicillium species were isolated, with 27 species being identified. The most common species were Penicillium polonicum (19.6 %), P. rubens (11.4 %), P. chrysogenum (11.4 %), and P. crustosum (10.9 %). The diversity of Penicillium strains isolated from soil (foreshore soil and sand) and marine macroorganisms was higher than the diversity of strains isolated from seawater. While many of the isolated strains showed alginase and β-glucosidase activity, no endoglucanase activity was found. More than half the strains (50.5 %) showed antifungal activity against at least one of the plant pathogens tested. Compared with other strains in this study, P. citrinum (strain SFC20140101-M662) showed high antifungal activity against both plant pathogens. The results reported here expand our knowledge of marine-derived Penicillium diversity. The relatively high proportion of strains that showed antifungal and enzyme activity demonstrates that marine-derived Penicillium have great potential to be used in the production of natural bioactive products for pharmaceutical and/or industrial use.

  2. A New record of four Penicillium species isolated from Agarum clathratum in Korea.

    PubMed

    Park, Myung Soo; Lee, Seobihn; Lim, Young Woon

    2017-04-01

    Agarum clathratum, brown algae, play important ecological roles in marine ecosystem, but can cause secondary environment pollution when they pile up on the beach. In order to resolve the environment problem by A. clathratum, we focus to isolate and identify Penicillium because many species are well known to produce extracellular enzymes. A total of 32 Penicillium strains were isolated from A. clathratum samples that collected from 13 sites along the mid-east coast of Korea in summer. They were identified based on morphological characters and phylogenetic analysis using β-tubulin DNA sequences as well as a combined dataset of β-tubulin and calmodulin. A total of 32 strains were isolated and they were identified to 13 Penicillium species. The commonly isolated species were Penicillium citrinum, P. roseomaculatum, and Penicillium sp. Among 13 Penicillium species, four species - P. bilaiae, P. cremeogriseum, P. madriti, and P. roseomaculatum - have not been previously recorded in Korea. For these four new species records to Korea, we provide morphological characteristics of each strain.

  3. Influence of storage temperature on growth of Penicillium polonicum and Penicillium glabrum and potential for deterioration of frozen chicken nuggets.

    PubMed

    Saccomori, Fernanda; Wigmann, Évelin Francine; Bernardi, Angélica Olivier; Alcano-González, María de Jesús; Copetti, Marina Venturini

    2015-05-04

    The practice of freezing food is one of the main processes used by the industry to prolong the shelf life of foods. Its use has expanded in recent years due to the increased consumption of convenience products, many of which are sold in frozen form. The temperature at which these foods are maintained during marketing in supermarkets or stored in the consumer's home is critical to ensure microbiological stability of products. Temperature abuse can allow microbial growth, especially growth of filamentous psychrophilic fungi. Besides economic losses in the industrial sector due to the return of products and loss of confidence by consumers, the development of fungi in foods is a public health problem due to the possibility of mycotoxin production. The aim of this study was to assess the growth at temperatures of 5, 0, -5 and -18°C for two species of fungi involved in the deterioration of frozen chicken nuggets, Penicillium polonicum (33/12 NGT) and Penicillium glabrum (29/12 NGT), inoculated both in culture medium and in the food. The results demonstrated that P. polonicum was able to form microcolonies on potato dextrose agar plates at 0°C and form visible colonies on the surface of the frozen chicken nuggets kept at -5°C for 120 days, regardless of brand. For P. glabrum the limiting growth temperature was 5°C in the culture medium and 0°C on frozen chicken nuggets, regardless of the brand analyzed. Thus, it is essential to adhere to the storage temperatures recommended to ensure the stability and safety of this food product.

  4. Inactivation of Penicillium digitatum and Penicillium italicum under in vitro and in vivo conditions by using UV-C light.

    PubMed

    Gündüz, Gülten Tıryakı; Pazir, Fikret

    2013-10-01

    In this study, the effects of UV-C on two of the main wound pathogens of citrus fruits, Penicillium digitatum and Penicillium italicum, were investigated with different inoculation methods in vitro and on oranges. P. digitatum and P. italicum spores were inoculated onto the surface of potato dextrose agar or oranges using spread, spot, wound, and piercing inoculation methods. UV-C treatment for 1 min from a working distance of 8 cm reduced the numbers of P. italicum and P. digitatum by about 3.9 and 5.3 log units, respectively, following spread inoculation under in vitro conditions. Significant reductions were obtained after 1-min UV-C treatments of the tested fungi following inoculation using the spread and spot methods. With inoculation by the wound and piercing methods, the tested spores were not inactivated completely even after 10- and 20-min treatment times, respectively. The application of UV-C (7.92 kJ m(-2)) on oranges reduced the percentage of oranges infected at least threefold compared with the rate of infection in the untreated control samples. UV-C irradiation could effectively inactivate spores of P. italicum and P. digitatum inoculated by the spread plate and spot inoculation methods under in vitro and in vivo conditions. On the other hand, because of the low penetration ability of UV-C light, the tested fungi were not completely inactivated following inoculation with the wound and piercing methods. UV-C treatment has potential for use in surface decontamination of citrus fruits.

  5. PCR-RFLP of ITS rDNA for the rapid identification of Penicillium subgenus Biverticillium species.

    PubMed

    Dupont, Jöelle; Dennetière, Bruno; Jacquet, Claire; Dupont, Marie France

    2006-09-01

    RFLP of ITS rDNA is proposed as a useful tool for molecular identification of the most common species of biverticillate penicillia. 60 isolates were analysed representing 13 species and 21 unique sequences were produced. The combination of five restriction enzymes was successful in separating 12 species. However, the variety Penicillium purpurogenum var. rubrisclerotium remained indistinguishable from Penicillium funiculosum. P. funiculosum appeared as the most confused species, being mis-identified with Penicillium miniolutum and Penicillium pinophilum, which were originally part of the species, and with P. purpurogenum perhaps because of the common production of red pigment. Penicillium variabile was difficult to investigate as introns were found on half of the isolates. Penicillium piceum, Penicillium rugulosum, Penicillium loliense, Penicillium erythromellis and P. purpurogenum were homogeneous from molecular and morphological positions and corresponded to a well circumscribed taxon. Furthermore, intraspecific variability was evidenced within P. pinophilum and P. funiculosum. The ex-type isolate of P. funiculosum produced a unique pattern. The method is sensitive, rapid and inexpensive and can be used for isolate identification of the biverticillate species. It is recommended particularly when many isolates have to be authentificated prior to analysis for phylogenetic assessment or population genetics.

  6. Penicillium salamii, a new species occurring during seasoning of dry-cured meat.

    PubMed

    Perrone, Giancarlo; Samson, Robert A; Frisvad, Jens C; Susca, Antonia; Gunde-Cimerman, Nina; Epifani, Filomena; Houbraken, Jos

    2015-01-16

    Fungi have an important role in the production of dry-cured meat products, especially during the seasoning period. In general, both industrially and handmade salami are quickly colonized by a composite mycobiota during seasoning, often with a strong predominance of Penicillium species. These species are involved in the improvement of the characteristics and taste, and in the prevention of the growth of pathogenic, toxigenic or spoilage fungi. During the survey of fungal species occurring on the salami surface and in the air of the seasoning and storage areas of a salami plant (Calabria, Italy), two Penicillium species were predominantly present. One species was identified as Penicillium nalgiovense, and the other was related to, but distinct from, Penicillium olsonii. Further molecular and biochemical analyses showed that this strain has high homology with the not yet described species named "Penicillium milanense" isolated in Denmark and Slovenia on cured meats. The taxonomic position of these strains in Penicillium was investigated using calmodulin, β tubulin and ITS sequences, phenotypic characters and extrolite patterns, and resulted in the discovery of a new Penicillium species, described here as P. salamii. A literature search showed that this species occurs on (cured) meat products worldwide. In our study, P. salamii predominated the salami and capocollo surface in levels similar to the commonly known starter culture P. nalgiovense, irrespective of the room or age of seasoning. Preliminary inoculation trials with P. salamii showed that it was able to colonize salami during seasoning, indicating that this species could be used as a fungal starter for dry-cured meat.

  7. Taxonomy, chemodiversity, and chemoconsistency of Aspergillus, Penicillium, and Talaromyces species

    PubMed Central

    Frisvad, Jens C.

    2014-01-01

    Aspergillus, Penicillium, and Talaromyces are among the most chemically inventive of all fungi, producing a wide array of secondary metabolites (exometabolites). The three genera are holophyletic in a cladistic sense and polythetic classes in an anagenetic or functional sense, and contain 344, 354, and 88 species, respectively. New developments in classification, cladification, and nomenclature have meant that the species, series, and sections suggested are natural groups that share many extrolites, including exometabolites, exoproteins, exocarbohydrates, and exolipids in addition to morphological features. The number of exometabolites reported from these species is very large, and genome sequencing projects have shown that a large number of additional exometabolites may be expressed, given the right conditions (“cryptic” gene clusters for exometabolites). The exometabolites are biosynthesized via shikimic acid, tricarboxylic acid cycle members, nucleotides, carbohydrates or as polyketides, non-ribosomal peptides, terpenes, or mixtures of those. The gene clusters coding for these compounds contain genes for the biosynthetic building blocks, the linking of these building blocks, tailoring enzymes, resistance for own products, and exporters. Species within a series or section in Aspergillus, Penicillium, and Talaromyces have many exometabolites in common, seemingly acquired by cladogenesis, but some the gene clusters for autapomorphic exometabolites may have been acquired by horizontal gene transfer. Despite genome sequencing efforts, and the many breakthroughs these will give, it is obvious that epigenetic factors play a large role in evolution and function of chemodiversity, and better methods for characterizing the epigenome are needed. Most of the individual species of the three genera produce a consistent and characteristic profile of exometabolites, but growth medium variations, stimulation by exometabolites from other species, and variations in abiotic

  8. Molecular characterization of patulin producing and non-producing Penicillium species in apples from Morocco.

    PubMed

    Rharmitt, Sanae; Hafidi, Majida; Hajjaj, Hassan; Scordino, Fabio; Giosa, Domenico; Giuffrè, Letterio; Barreca, Davide; Criseo, Giuseppe; Romeo, Orazio

    2016-01-18

    The isolation of patulin-producing Penicillia in apples collected in different markets in four localities in Morocco is reported. Fungi were identified by β-tubulin sequencing and further characterized using a specific PCR-based method targeting the isoepoxydon dehydrogenase (IDH) gene to discriminate between patulin-producing and non-producing strains. Production of patulin was also evaluated using standard cultural and biochemical methods. Results showed that 79.5% of contaminant fungi belonged to the genus Penicillium and that Penicillium expansum was the most isolated species (83.9%) followed by Penicillium chrysogenum (~9.7%) and Penicillium crustosum (~6.4%). Molecular analysis revealed that 64.5% of the Penicillium species produced the expected IDH-amplicon denoting patulin production in these strains. However, patulin production was not chemically confirmed in all P. expansum strains. The isolation of IDH(-)/patulin(+) strains poses the hypothesis that gentisylaldehyde is not a direct patulin precursor, supporting previous observations that highlighted the importance of the gentisyl alcohol in the production of this mycotoxin. Total agreement between IDH-gene detection and cultural/chemical methods employed was observed in 58% of P. expansum strains and for 100% of the other species isolated. Overall the data reported here showed a substantial genetic variability within P. expansum population from Morocco.

  9. Genomic and secretomic analyses reveal unique features of the lignocellulolytic enzyme system of Penicillium decumbens.

    PubMed

    Liu, Guodong; Zhang, Lei; Wei, Xiaomin; Zou, Gen; Qin, Yuqi; Ma, Liang; Li, Jie; Zheng, Huajun; Wang, Shengyue; Wang, Chengshu; Xun, Luying; Zhao, Guo-Ping; Zhou, Zhihua; Qu, Yinbo

    2013-01-01

    Many Penicillium species could produce extracellular enzyme systems with good lignocellulose hydrolysis performance. However, these species and their enzyme systems are still poorly understood and explored due to the lacking of genetic information. Here, we present the genomic and secretomic analyses of Penicillium decumbens that has been used in industrial production of lignocellulolytic enzymes in China for more than fifteen years. Comparative genomics analysis with the phylogenetically most similar species Penicillium chrysogenum revealed that P. decumbens has evolved with more genes involved in plant cell wall degradation, but fewer genes in cellular metabolism and regulation. Compared with the widely used cellulase producer Trichoderma reesei, P. decumbens has a lignocellulolytic enzyme system with more diverse components, particularly for cellulose binding domain-containing proteins and hemicellulases. Further, proteomic analysis of secretomes revealed that P. decumbens produced significantly more lignocellulolytic enzymes in the medium with cellulose-wheat bran as the carbon source than with glucose. The results expand our knowledge on the genetic information of lignocellulolytic enzyme systems in Penicillium species, and will facilitate rational strain improvement for the production of highly efficient enzyme systems used in lignocellulose utilization from Penicillium species.

  10. Genomic and Secretomic Analyses Reveal Unique Features of the Lignocellulolytic Enzyme System of Penicillium decumbens

    PubMed Central

    Qin, Yuqi; Ma, Liang; Li, Jie; Zheng, Huajun; Wang, Shengyue; Wang, Chengshu; Xun, Luying; Zhao, Guo-Ping; Zhou, Zhihua; Qu, Yinbo

    2013-01-01

    Many Penicillium species could produce extracellular enzyme systems with good lignocellulose hydrolysis performance. However, these species and their enzyme systems are still poorly understood and explored due to the lacking of genetic information. Here, we present the genomic and secretomic analyses of Penicillium decumbens that has been used in industrial production of lignocellulolytic enzymes in China for more than fifteen years. Comparative genomics analysis with the phylogenetically most similar species Penicillium chrysogenum revealed that P. decumbens has evolved with more genes involved in plant cell wall degradation, but fewer genes in cellular metabolism and regulation. Compared with the widely used cellulase producer Trichoderma reesei, P. decumbens has a lignocellulolytic enzyme system with more diverse components, particularly for cellulose binding domain-containing proteins and hemicellulases. Further, proteomic analysis of secretomes revealed that P. decumbens produced significantly more lignocellulolytic enzymes in the medium with cellulose-wheat bran as the carbon source than with glucose. The results expand our knowledge on the genetic information of lignocellulolytic enzyme systems in Penicillium species, and will facilitate rational strain improvement for the production of highly efficient enzyme systems used in lignocellulose utilization from Penicillium species. PMID:23383313

  11. Secondary Metabolites from Penicillium roqueforti, A Starter for the Production of Gorgonzola Cheese

    PubMed Central

    Giardini, Alberto; Soncini, Gabriella

    2014-01-01

    The presence of mold in food, although necessary for production, can involve the presence of secondary metabolites, which are sometimes toxic. Penicillium roqueforti is a common saprophytic fungus but it is also the essential fungus used in the production of Roquefort cheese and other varieties of blue cheese containing internal mold. The study was conducted on industrial batches of Penicillium roqueforti starters used in the production of the Gorgonzola cheese, with the aim to verify the production of secondary metabolites. Nine Penicillium roqueforti strains were tested. The presence of roquefortine C, PR toxin and mycophenolic acid was tested first in vitro, then on bread-like substrate and lastly in vivo in nine cheese samples produced with the same starters and ready to market. In vitro, only Penicillium out of nine produced roquefortine C, four starters showed mycophenolic acid production, while no significant amounts of PR toxin were detected. In the samples grown on bread-like substrate, Penicillium did not produce secondary metabolites, likewise with each cheese samples tested. To protect consumers’ health and safety, the presence of mycotoxins needs to be verified in food which is widely consumed, above all for products protected by the protected denomination of origin (DOP) label (i.e. a certificate guaranteeing the geographic origin of the product), such as Gorgonzola cheese. PMID:27800360

  12. Patulin and secondary metabolite production by marine-derived Penicillium strains.

    PubMed

    Vansteelandt, Marieke; Kerzaon, Isabelle; Blanchet, Elodie; Fossi Tankoua, Olivia; Robiou Du Pont, Thibaut; Joubert, Yolaine; Monteau, Fabrice; Le Bizec, Bruno; Frisvad, Jens C; Pouchus, Yves François; Grovel, Olivier

    2012-09-01

    Genus Penicillium represents an important fungal group regarding to its mycotoxin production. Secondary metabolomes of eight marine-derived strains belonging to subgenera Furcatum and Penicillium were investigated using dereplication by liquid chromatography (LC)-Diode Array Detector (DAD)-mass spectrometry (MS)/MS. Each strain was grown on six different culture media to enhance the number of observable metabolites. Thirty-two secondary metabolites were detected in crude extracts with twenty first observations for studied species. Patulin, a major mycotoxin, was classically detected in extracts of Penicillium expansum, and was also isolated from Penicillium antarcticum cultures, whose secondary metabolome is still to be done. These detections constituted the first descriptions of patulin in marine strains of Penicillium, highlighting the risk for shellfish and their consumers due to the presence of these fungi in shellfish farming areas. Patulin induced acute neurotoxicity on Diptera larvae, indicating the interest of this bioassay as an additional tool for detection of this major mycotoxin in crude extracts.

  13. Identification and Antifungal Susceptibility of Penicillium-Like Fungi from Clinical Samples in the United States

    PubMed Central

    Guevara-Suarez, Marcela; Sutton, Deanna A.; García, Dania; Martin-Vicente, Adela; Wiederhold, Nathan; Guarro, Josep; Gené, Josepa

    2016-01-01

    Penicillium species are some of the most common fungi observed worldwide and have an important economic impact as well as being occasional agents of human and animal mycoses. A total of 118 isolates thought to belong to the genus Penicillium based on morphological features were obtained from the Fungus Testing Laboratory at the University of Texas Health Science Center in San Antonio (United States). The isolates were studied phenotypically using standard growth conditions. Molecular identification was made using two genetic markers, the internal transcribed spacer (ITS) and a fragment of the β-tubulin gene. In order to assess phylogenetic relationships, maximum likelihood and Bayesian inference assessments were used. Antifungal susceptibility testing was performed according to CLSI document M38-A2 for nine antifungal drugs. The isolates were identified within three genera, i.e., Penicillium, Talaromyces, and Rasamsonia. The most frequent species in our study were Penicillium rubens, P. citrinum, and Talaromyces amestolkiae. The potent in vitro activity of amphotericin B (AMB) and terbinafine (TRB) and of the echinocandins against Penicillium and Talaromyces species might offer a good therapeutic alternative for the treatment of infections caused by these fungi. PMID:27280422

  14. Improvement of strain Penicillium sp. EZ-ZH190 for tannase production by induced mutation.

    PubMed

    Zakipour-Molkabadi, E; Hamidi-Esfahani, Z; Sahari, M A; Azizi, M H

    2013-11-01

    In the search for an efficient producer of tannase, Penicillium sp. EZ-ZH190 was subjected to mutagenesis using heat treatment and strain EZ-ZH290 was isolated. The maximum tannase in this mutant strain was 4.32 U/mL with an incubation period of 84 h as compared to wild strain EZ-ZH190 where the incubation period was 96 h with a maximum enzyme activity of 4.33 U/mL. Also, the Penicillium sp. EZ-ZH290 tannase had a maximum activity at 40 °C and pH 5.5. Then, the spores of strain EZ-ZH290 were subjected to γ irradiation mutagenesis and strain EZ-ZH390 was isolated. Strain EZ-ZH390 exhibited higher tannase activity (7.66 U/mL) than the parent strain EZ-ZH290. It was also found that Penicillium sp. EZ-ZH390 tannase had an optimum activity at 35 °C and a broad pH profile with an optimum at pH 5.5. The tannase pH stability of Penicillium sp. EZ-ZH390 and its maximum production of tannase followed the same trend for five generations confirming the occurrence of stable mutant. This paper is shown that γ irradiation can mutate the Penicillium sp. leading to increase the tannase production.

  15. Identification and Antifungal Susceptibility of Penicillium-Like Fungi from Clinical Samples in the United States.

    PubMed

    Guevara-Suarez, Marcela; Sutton, Deanna A; Cano-Lira, José F; García, Dania; Martin-Vicente, Adela; Wiederhold, Nathan; Guarro, Josep; Gené, Josepa

    2016-08-01

    Penicillium species are some of the most common fungi observed worldwide and have an important economic impact as well as being occasional agents of human and animal mycoses. A total of 118 isolates thought to belong to the genus Penicillium based on morphological features were obtained from the Fungus Testing Laboratory at the University of Texas Health Science Center in San Antonio (United States). The isolates were studied phenotypically using standard growth conditions. Molecular identification was made using two genetic markers, the internal transcribed spacer (ITS) and a fragment of the β-tubulin gene. In order to assess phylogenetic relationships, maximum likelihood and Bayesian inference assessments were used. Antifungal susceptibility testing was performed according to CLSI document M38-A2 for nine antifungal drugs. The isolates were identified within three genera, i.e., Penicillium, Talaromyces, and Rasamsonia The most frequent species in our study were Penicillium rubens, P. citrinum, and Talaromyces amestolkiae The potent in vitro activity of amphotericin B (AMB) and terbinafine (TRB) and of the echinocandins against Penicillium and Talaromyces species might offer a good therapeutic alternative for the treatment of infections caused by these fungi.

  16. Identification of Penicillium marneffei in Paraffin-Embedded Tissue Using Nested PCR.

    PubMed

    Zeng, Hanxiang; Li, Xiqing; Chen, Xiejie; Zhang, Junmin; Sun, Jiufeng; Xie, Zhi; Xi, Liyan

    2009-07-01

    Penicillium marneffei is one of the unique thermally dimorphic fungi in Penicillium species that causes a disseminated, progressive and life threatening infection in immunocompromised patients. The diagnosis of Penicilliosis marneffei depends on culture that may delay the treatment due to the time-consuming process. In the present study, we evaluated the specificity and sensitivity of nested PCR to identify Penicillium marneffei from paraffin-embedded tissue. Two sets of oligonucleotide primers were derived from the sequence of 18S rRNA of Penicillium marneffei. The outer primers (RRF1 and RRH1) were specific to fungi. The inner primers (Pm1 and Pm2) were specific to Penicillium marneffei. The specific fragment of approximately 400 bp was amplified from all paraffin-embedded tissues from 14 patients with Penicilliosis marneffei and 10 bamboo rats. The detectable DNA concentration of single PCR and nested PCR were 14 pg/microl and 14 fg/microl, respectively. Further studies are required in order to use nested PCR for early diagnosis of the disease.

  17. CRISPR/Cas9 Based Genome Editing of Penicillium chrysogenum.

    PubMed

    Pohl, C; Kiel, J A K W; Driessen, A J M; Bovenberg, R A L; Nygård, Y

    2016-07-15

    CRISPR/Cas9 based systems have emerged as versatile platforms for precision genome editing in a wide range of organisms. Here we have developed powerful CRISPR/Cas9 tools for marker-based and marker-free genome modifications in Penicillium chrysogenum, a model filamentous fungus and industrially relevant cell factory. The developed CRISPR/Cas9 toolbox is highly flexible and allows editing of new targets with minimal cloning efforts. The Cas9 protein and the sgRNA can be either delivered during transformation, as preassembled CRISPR-Cas9 ribonucleoproteins (RNPs) or expressed from an AMA1 based plasmid within the cell. The direct delivery of the Cas9 protein with in vitro synthesized sgRNA to the cells allows for a transient method for genome engineering that may rapidly be applicable for other filamentous fungi. The expression of Cas9 from an AMA1 based vector was shown to be highly efficient for marker-free gene deletions.

  18. New insights into the isopenicillin N transport in Penicillium chrysogenum.

    PubMed

    Fernández-Aguado, M; Martín, J F; Rodríguez-Castro, R; García-Estrada, C; Albillos, S M; Teijeira, F; Ullán, R V

    2014-03-01

    In Penicillium chrysogenum the beta-lactam biosynthetic pathway is compartmentalized. This fact forces the occurrence of transport processes of penicillin-intermediate molecules across cell membranes. Many aspects around this molecular traffic remain obscure but are supposed to involve transmembrane transporter proteins. In the present work, an in-depth study has been developed on a Major Facilitator-type secondary transporter from P. chrysogenum named as PenM. The reduction of penM expression level reached by penM targeted silencing, leads to a decrease in benzylpenicillin production in silenced transformants, especially in SilM-35. On the contrary, the penM overexpression from a high efficiency promoter increases the benzylpenicillin production and the expression of the biosynthetic genes. Moreover, when the silenced strain SilM-35 is cultured under penicillin production conditions with 6-aminopenicillanic acid supplementation, an increase in the benzylpenicillin production proportional to the 6-aminopenicillanic acid availability is observed. By this phenomenon, it can be concluded that due to the penM silencing the benzylpenicillin transport remains intact but the peroxisomal isopenicillin N import results affected. As a culminating result, obtained by the expression of the fluorescent recombinant PenM-DsRed protein, it was determined that PenM is naturally located in P. chrysogenum peroxisomes. In summary, our experimental results suggest that PenM is involved in penicillin production most likely through the translocation of isopenicillin N from the cytosol to the peroxisomal lumen across P. chrysogenum peroxisomal membrane.

  19. Peroxisomes are required for efficient penicillin biosynthesis in Penicillium chrysogenum.

    PubMed

    Meijer, Wiebe H; Gidijala, Loknath; Fekken, Susan; Kiel, Jan A K W; van den Berg, Marco A; Lascaris, Romeo; Bovenberg, Roel A L; van der Klei, Ida J

    2010-09-01

    In the fungus Penicillium chrysogenum, penicillin (PEN) production is compartmentalized in the cytosol and in peroxisomes. Here we show that intact peroxisomes that contain the two final enzymes of PEN biosynthesis, acyl coenzyme A (CoA):6-amino penicillanic acid acyltransferase (AT) as well as the side-chain precursor activation enzyme phenylacetyl CoA ligase (PCL), are crucial for efficient PEN synthesis. Moreover, increasing PEN titers are associated with increasing peroxisome numbers. However, not all conditions that result in enhanced peroxisome numbers simultaneously stimulate PEN production. We find that conditions that lead to peroxisome proliferation but simultaneously interfere with the normal physiology of the cell may be detrimental to antibiotic production. We furthermore show that peroxisomes develop in germinating conidiospores from reticule-like structures. During subsequent hyphal growth, peroxisome proliferation occurs at the tip of the growing hyphae, after which the organelles are distributed over newly formed subapical cells. We observed that the organelle proliferation machinery requires the dynamin-like protein Dnm1.

  20. Safety evaluation of glucose oxidase from Penicillium chrysogenum.

    PubMed

    Konishi, Tetsuya; Aoshima, Takuya; Mizuhashi, Fukutaro; Choi, Sharon S H; Roberts, Ashley

    2013-06-01

    Glucose oxidase (β-d-glucose:oxygen 1-oxidoreductase; EC 1.1.2.3.4) is used in the food and beverage industry as a preservative and stabilizer and is commonly derived from the fungus Aspergillus niger. Although the safety of glucose oxidase preparations from A. niger is well-established, the use of preparations derived from other fungal species is of interest; however, an assessment of their safety is warranted. Here, we report on the safety of a glucose oxidase preparation derived from the fungus Penicillium chrysogenum (designated as PGO) for commercial use in food processing, as well as an ingredient in food. In a repeated dose 90-day oral toxicity study conducted in rats, PGO was without compound-related adverse effects at doses of up to 15,600U/kg body weight/day, equivalent to 193mg total organic solids/kg body weight/day. In addition, PGO was non-genotoxic in a series of genotoxicity tests, including a bacterial reverse mutation test, an in vitro mammalian chromosomal aberration test, and a combined in vivo mammalian erythrocyte micronucleus test and comet assay. The results of these studies support the safe use of PGO in food for human consumption.

  1. Scale-down of penicillin production in Penicillium chrysogenum.

    PubMed

    de Jonge, Lodewijk P; Buijs, Nicolaas A A; ten Pierick, Angela; Deshmukh, Amit; Zhao, Zheng; Kiel, Jan A K W; Heijnen, Joseph J; van Gulik, Walter M

    2011-08-01

    In large-scale production reactors the combination of high broth viscosity and large broth volume leads to insufficient liquid-phase mixing, resulting in gradients in, for example, the concentrations of substrate and oxygen. This often leads to differences in productivity of the full-scale process compared with laboratory scale. In this scale-down study of penicillin production, the influence of substrate gradients on process performance and cell physiology was investigated by imposing an intermittent feeding regime on a laboratory-scale culture of a high yielding strain of Penicillium chrysogenum. It was found that penicillin production was reduced by a factor of two in the intermittently fed cultures relative to constant feed cultivations fed with the same amount of glucose per hour, while the biomass yield was the same. Measurement of the levels of the intermediates of the penicillin biosynthesis pathway, along with the enzyme levels, suggested that the reduction of the flux through the penicillin pathway is mainly the result of a lower influx into the pathway, possibly due to inhibitory levels of adenosine monophosphate and pyrophosphate and lower activating levels of adenosine triphosphate during the zero-substrate phase of each cycle of intermittent feeding.

  2. Potential of Penicillium Species in the Bioremediation Field

    PubMed Central

    Leitão, Ana Lúcia

    2009-01-01

    The effects on the environment of pollution, particularly that caused by various industrial activities, have been responsible for the accelerated fluxes of organic and inorganic matter in the ecosphere. Xenobiotics such as phenol, phenolic compounds, polycyclic aromatic hydrocarbons (PAHs), and heavy metals, even at low concentrations, can be toxic to humans and other forms of life. Many of the remediation technologies currently being used for contaminated soil and water involve not only physical and chemical treatment, but also biological processes, where microbial activity is the responsible for pollutant removal and/or recovery. Fungi are present in aquatic sediments, terrestrial habitats and water surfaces and play a significant part in natural remediation of metal and aromatic compounds. Fungi also have advantages over bacteria since fungal hyphae can penetrate contaminated soil, reaching not only heavy metals but also xenobiotic compounds. Despite of the abundance of such fungi in wastes, penicillia in particular have received little attention in bioremediation and biodegradation studies. Additionally, several studies conducted with different strains of imperfecti fungi, Penicillium spp. have demonstrated their ability to degrade different xenobiotic compounds with low co-substrate requirements, and could be potentially interesting for the development of economically feasible processes for pollutant transformation. PMID:19440525

  3. Penicillium expansum volatiles reduce pine weevil attraction to host plants.

    PubMed

    Azeem, Muhammad; Rajarao, Gunaratna Kuttuva; Nordenhem, Henrik; Nordlander, Göran; Borg-Karlson, Anna Karin

    2013-01-01

    The pine weevil Hylobius abietis (L.) is a severe pest of conifer seedlings in reforested areas of Europe and Asia. To identify minimally toxic and ecologically sustainable compounds for protecting newly planted seedlings, we evaluated the volatile metabolites produced by microbes isolated from H. abietis feces and frass. Female weevils deposit feces and chew bark at oviposition sites, presumably thus protecting eggs from feeding conspecifics. We hypothesize that microbes present in feces/frass are responsible for producing compounds that deter weevils. Here, we describe the isolation of a fungus from feces and frass of H. abietis and the biological activity of its volatile metabolites. The fungus was identified by morphological and molecular methods as Penicillium expansum Link ex. Thom. It was cultured on sterilized H. abietis frass medium in glass flasks, and volatiles were collected by SPME and analyzed by GC-MS. The major volatiles of the fungus were styrene and 3-methylanisole. The nutrient conditions for maximum production of styrene and 3-methylanisole were examined. Large quantities of styrene were produced when the fungus was cultured on grated pine bark with yeast extract. In a multi-choice arena test, styrene significantly reduced male and female pine weevils' attraction to cut pieces of Scots pine twigs, whereas 3-methylanisole only reduced male weevil attraction to pine twigs. These studies suggest that metabolites produced by microbes may be useful as compounds for controlling insects, and could serve as sustainable alternatives to synthetic insecticides.

  4. Inhibition of Penicillium expansum by an oxidative treatment.

    PubMed

    Cerioni, Luciana; Lazarte, María de Los Ángeles; Villegas, Josefina María; Rodríguez-Montelongo, Luisa; Volentini, Sabrina Inés

    2013-04-01

    Several oxidizing compounds such as sodium hypochlorite (NaClO) and hydrogen peroxide (H(2)O(2)) are used to control postharvest decay in fresh fruit due to their antimicrobial effects. Here, we applied these compounds in vitro, in the presence of CuSO(4), against Penicillium expansum, causal agent of apple blue mold. MICs were 50 mg L(-1) and 400 mmol L(-1) for NaClO and H(2)O(2), respectively, when these compounds were individually applied to conidia suspensions during 2 min. A combined oxidative treatment (OT) consisting on an incubation with 1 mg L(-1) NaClO and 200 mmol L(-1) H(2)O(2), in the presence of 6 mmol L(-1) CuSO(4), inhibited growth, conidial germination and fungal infectivity on apple. The fractional inhibitory concentration index for the interaction between NaClO and H(2)O(2) in the OT was 0.52 indicating a synergistic effect of the oxidizing compounds. These results suggest that the OT could be an interesting alternative for apple diseases postharvest control.

  5. Ecophysiological characterization of Penicillium expansum population in lleida (Spain).

    PubMed

    Morales, Hector; Marín, Sonia; Obea, Laura; Patiño, Belén; Doménech, Miriam; Ramos, Antonio J; Sanchis, Vicente

    2008-03-20

    Penicillium expansum, a patulin producer fungus, is the most important fungus causing decay in cold stored both apples and pears. This can lead to patulin contaminated by-products. The aim of this assay was to evaluate the phenotypical and physiological variability in the population of P. expansum that cause fruit spoilage in post-harvest stages in Lleida (Spain). In total, 101 isolates of P. expansum from the 2004 and the 2005 seasons were obtained from decayed fruits. Significant differences were found in the observations from both seasons. Variability of the isolates in each season seemed to be partially explained by differences in growth in media, patulin accumulation and resistance to fungicides. Patulin production was detected in almost 100% of the isolates. Variability existing in P. expansum population could not be totally explained, but the above mentioned variables explained up to 74% of the diversity in some cases. The results obtained point to the existence of different populations of P. expansum in each season and may explain the differences in fungicide resistance observed between both seasons. The capacity to colonize apple flesh and some variables involved in fruit colonization were not a source of variation neither in each season nor when both seasons were compared. As storage rooms are cleaned and disinfected each season, this suggests that each season, the populations in storage rooms develop only from strains capable to colonize apple flesh. This may lead to rapid sporulation and spreading of spores.

  6. Kinetics of biofilm formation by drinking water isolated Penicillium expansum.

    PubMed

    Simões, Lúcia Chaves; Simões, Manuel; Lima, Nelson

    2015-01-01

    Current knowledge on drinking water (DW) biofilms has been obtained mainly from studies on bacterial biofilms. Very few reports on filamentous fungi (ff) biofilms are available, although they can contribute to the reduction in DW quality. This study aimed to assess the dynamics of biofilm formation by Penicillium expansum using microtiter plates under static conditions, mimicking water flow behaviour in stagnant regions of drinking water distribution systems. Biofilms were analysed in terms of biomass (crystal violet staining), metabolic activity (resazurin, fluorescein diacetate and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide [MTT]) and morphology (epifluorescence [calcofluor white M2R, FUN-1, FDA and acridine orange] and bright-field microscopies). Biofilm development over time showed the typical sigmoidal curve with noticeable different phases in biofilm formation (induction, exponential, stationary, and sloughing off). The methods used to assess metabolic activity provided similar results. The microscope analysis allowed identification of the involvement of conidia in initial adhesion (4 h), germlings (8 h), initial monolayers (12 h), a monolayer of intertwined hyphae (24 h), mycelial development, hyphal layering and bundling, and development of the mature biofilms (≥48 h). P. expansum grows as a complex, multicellular biofilm in 48 h. The metabolic activity and biomass of the fungal biofilms were shown to increase over time and a correlation between metabolism, biofilm mass and hyphal development was found.

  7. Photodynamic inactivation of Penicillium chrysogenum conidia by cationic porphyrins.

    PubMed

    Gomes, Maria C; Woranovicz-Barreira, Sandra M; Faustino, Maria A F; Fernandes, Rosa; Neves, Maria G P M S; Tomé, Augusto C; Gomes, Newton C M; Almeida, Adelaide; Cavaleiro, José A S; Cunha, Angela; Tomé, João P C

    2011-11-01

    This work reports the photophysical and biological evaluation of five cationic porphyrins as photosensitizers (PS) for the photodynamic inactivation (PDI) of Penicillium chrysogenum conidia. Two different cationic porphyrin groups were synthesized from 5,10,15,20-tetrakis(4-pyridyl)porphyrin and 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin. The photostability and singlet oxygen generation studies showed that these molecules are photostable and efficient singlet oxygen generators. PDI experiments of P. chrysogenum conidia conducted with 50 μmol L(-1) of photosensitiser under white light at a fluence rate of 200 mW cm(-2) over 20 min showed that the most effective PS caused a 4.1 log reduction in the concentration of viable conidia. The present results show that porphyrins 1a and 1b are more efficient PSs than porphyrin 2a while porphyrins 1c and 2b show no inactivation of P. chrysogenum. It is also clear that the effectiveness of the molecule as PS for antifungal PDI is strongly related with the porphyrin substituent groups, and consequently their solubility in physiological media. The average amount of PS adsorbed per viable conidium was a determining factor in the photoinactivation efficiency and varied between the different studied PSs. Cationic PSs 1a and 1b might be promising anti-fungal PDI agents with potential applications in phytosanitation, biofilm control, bioremediation, and wastewater treatment.

  8. A cold-adapted endo-arabinanase from Penicillium chrysogenum.

    PubMed

    Sakamoto, T; Ihara, H; Kozaki, S; Kawasaki, H

    2003-12-05

    Previously, three arabinan-degrading enzymes were isolated from Penicillium chrysogenum 31B. Here we describe another arabinan-degrading enzyme, termed Abnc, from the culture filtrate of the same organism. Analysis of the reaction products of debranched arabinan by high-performance anion-exchange chromatography (HPAEC) revealed that Abnc cleaved the substrate in an endo manner and that the final major product was arabinotriose. The molecular mass of Abnc was estimated to be 35 kDa by SDS-PAGE. Enzyme activity of Abnc was highest at pH 6.0 to 7.0. The enzyme was stable up to 30 degrees C and showed optimum activity at 30 to 40 degrees C. Compared with a mesophilic counterpart from Aspergillus niger, Abnc exhibited a lower thermal stability and optimum enzyme activity at lower temperatures. Production of Abnc in P. chrysogenum was found to be strongly induced by arabinose-containing polymers and required a longer culture time than did other arabinanase isozymes in this strain.

  9. A taxonomic and phylogenetic revision of Penicillium section Aspergilloides

    PubMed Central

    Houbraken, J.; Visagie, C.M.; Meijer, M.; Frisvad, J.C.; Busby, P.E.; Pitt, J.I.; Seifert, K.A.; Louis-Seize, G.; Demirel, R.; Yilmaz, N.; Jacobs, K.; Christensen, M.; Samson, R.A.

    2014-01-01

    Species belonging to Penicillium section Aspergilloides have a world-wide distribution with P. glabrum, P. spinulosum and P. thomii the most well-known species of this section. These species occur commonly and can be isolated from many substrates including soil, food, bark and indoor environments. The taxonomy of these species has been investigated several times using various techniques, but species delimitation remains difficult. In the present study, 349 strains belonging to section Aspergilloides were subjected to multilocus molecular phylogenetic analyses using partial β-tubulin (BenA), calmodulin (CaM) and RNA polymerase II second largest subunit (RPB2) sequences. Section Aspergilloides is subdivided into 12 clades and 51 species. Twenty-five species are described here as new and P. yezoense, a species originally described without a Latin diagnosis, is validated. Species belonging to section Aspergilloides are phenotypically similar and most have monoverticillate conidiophores and grow moderately or quickly on agar media. The most important characters to distinguish these species were colony sizes on agar media, growth at 30 °C, ornamentation and shape of conidia, sclerotium production and stipe roughness. PMID:25492984

  10. Antifungal effect and mechanism of garlic oil on Penicillium funiculosum.

    PubMed

    Li, Wen-Ru; Shi, Qing-Shan; Liang, Qing; Huang, Xiao-Mo; Chen, Yi-Ben

    2014-10-01

    Garlic oil is a kind of fungicide, but little is known about its antifungal effects and mechanism. In this study, the chemical constituents, antifungal activity, and effects of garlic oil were studied with Penicillium funiculosum as a model strain. Results showed that the minimum fungicidal concentrations (MFCs, v/v) were 0.125 and 0.0313 % in agar medium and broth medium, respectively, suggesting that the garlic oil had a strong antifungal activity. The main ingredients of garlic oil were identified as sulfides, mainly including disulfides (36 %), trisulfides (32 %) and monosulfides (29 %) by gas chromatograph-mass spectrometer (GC/MS), which were estimated as the dominant antifungal factors. The observation results by transmission electron microscope (TEM) and scanning electron microscope (SEM) indicated that garlic oil could firstly penetrate into hyphae cells and even their organelles, and then destroy the cellular structure, finally leading to the leakage of both cytoplasm and macromolecules. Further proteomic analysis displayed garlic oil was able to induce a stimulated or weakened expression of some key proteins for physiological metabolism. Therefore, our study proved that garlic oil can work multiple sites of the hyphae of P. funiculosum to cause their death. The high antifungal effects of garlic oil makes it a broad application prospect in antifungal industries.

  11. Bioactive Chaetoglobosins from the Mangrove Endophytic Fungus Penicillium chrysogenum

    PubMed Central

    Huang, Song; Chen, Haiyan; Li, Wensheng; Zhu, Xinwei; Ding, Weijia; Li, Chunyuan

    2016-01-01

    A novel chaetoglobosin named penochalasin I (1) with a unprecedented six-cyclic 6/5/6/5/6/13 fused ring system, and another new chaetoglobosin named penochalasin J (2), along with chaetoglobosins G, F, C, A, E, armochaetoglobosin I, and cytoglobosin C (3–9) were isolated from the culture of Penicillium chrysogenum V11. Their structures were elucidated by 1D, 2D NMR spectroscopic analysis and high resolution mass spectroscopic data. The absolute configuration of compounds 1 and 2 were determined by comparing the theoretical electronic circular dichroism (ECD) calculation with the experimental CD. Compound 1 was the first example, with a six-cyclic fused ring system formed by the connection of C-5 and C-2′ of the chaetoglobosin class. Compounds 5–8 remarkably inhibited the plant pathogenic fungus R. solani (minimum inhibitory concentrations (MICs) = 11.79–23.66 μM), and compounds 2, 6, and 7 greatly inhibited C. gloeosporioides (MICs = 23.58–47.35 μM), showing an antifungal activity higher than that of carbendazim. Compound 1 exhibited marked cytotoxicity against MDA-MB-435 and SGC-7901 cells (IC50 < 10 μM), and compounds 6 and 9 showed potent cytotoxicity against SGC-7901 and A549 cells (IC50 < 10 μM). PMID:27690061

  12. Zosteropenillines: Polyketides from the Marine-Derived Fungus Penicillium thomii

    PubMed Central

    Afiyatullov, Shamil Sh.; Leshchenko, Elena V.; Berdyshev, Dmitrii V.; Sobolevskaya, Maria P.; Antonov, Alexandr S.; Denisenko, Vladimir A.; Popov, Roman S.; Pivkin, Mikhail V.; Udovenko, Anatoly A.; Pislyagin, Evgeny A.; von Amsberg, Gunhild; Dyshlovoy, Sergey A.

    2017-01-01

    Twelve new polyketides, zosteropenillines A–L (1–12), together with known polyketide pallidopenilline A (13), were isolated from the ethylacetate extract of the fungus Penicillium thomii associated with the seagrass Zostera marina. Their structures were established based on spectroscopic methods. The absolute configuration of zosteropenilline A (1) as 4R, 5S, 8S, 9R, 10R, and 13S was determined by a combination of the modified Mosher’s method, X-ray analysis, and NOESY data. Absolute configurations of zosteropenillines B–D (2–4) were determined by time-dependent density functional theory (TD-DFT) calculations of ECD spectra. The effect of compounds 1–3, 7, 8, 10, and 11 on the viability of human drug-resistant prostate cancer cells PC3 as well as on autophagy in these cancer cells and inhibitory effects of compounds 1, 2, and 8–10 on NO production in LPS-induced RAW 264.7 murine macrophages were examined. PMID:28218691

  13. The Penicillium echinulatum Secretome on Sugar Cane Bagasse

    PubMed Central

    Ribeiro, Daniela A.; Cota, Júnio; Alvarez, Thabata M.; Brüchli, Fernanda; Bragato, Juliano; Pereira, Beatriz M. P.; Pauletti, Bianca A.; Jackson, George; Pimenta, Maria T. B.; Murakami, Mario T.; Camassola, Marli; Ruller, Roberto; Dillon, Aldo J. P.; Pradella, Jose G. C.; Paes Leme, Adriana F.; Squina, Fabio M.

    2012-01-01

    Plant feedstocks are at the leading front of the biofuel industry based on the potential to promote economical, social and environmental development worldwide through sustainable scenarios related to energy production. Penicillium echinulatum is a promising strain for the bioethanol industry based on its capacity to produce large amounts of cellulases at low cost. The secretome profile of P. echinulatum after grown on integral sugarcane bagasse, microcrystalline cellulose and three types of pretreated sugarcane bagasse was evaluated using shotgun proteomics. The comprehensive chemical characterization of the biomass used as the source of fungal nutrition, as well as biochemical activity assays using a collection of natural polysaccharides, were also performed. Our study revealed that the enzymatic repertoire of P. echinulatum is geared mainly toward producing enzymes from the cellulose complex (endogluganases, cellobiohydrolases and β-glucosidases). Glycoside hydrolase (GH) family members, important to biomass-to-biofuels conversion strategies, were identified, including endoglucanases GH5, 7, 6, 12, 17 and 61, β-glycosidase GH3, xylanases GH10 and GH11, as well as debranching hemicellulases from GH43, GH62 and CE2 and pectinanes from GH28. Collectively, the approach conducted in this study gave new insights on the better comprehension of the composition and degradation capability of an industrial cellulolytic strain, from which a number of applied technologies, such as biofuel production, can be generated. PMID:23227186

  14. Safety evaluation of nuclease P1 from Penicillium citrinum.

    PubMed

    Okado, Nobuo; Hasegawa, Kazushige; Mizuhashi, Fukutaro; Lynch, Barry S; Vo, Trung D; Roberts, Ashley S

    2016-02-01

    Nuclease P1 has been widely used in the food industry to enhance or create flavor. One commercial source of this enzyme is Penicillium citrinum, an anamorphic mesophilic fungus with a long history of safe use in Europe and Asia as a fermentation organism used in the production of ribonucleases. Given the intended use in food for human consumption, and noting its potential presence at trace levels in finished products, a series of safety studies including an in vitro Ames and chromosome aberration assay, an in vivo rat erythrocyte micronucleus assay and a 90-day oral toxicity study in rats were conducted. No mutagenic activity was observed in the Ames assay. Equivocal activity in the chromosome aberration assay was not replicated in the micronucleus assay at doses of up to 1007 mg total organic solids (TOS)/kg body weight (bw)/day. Following oral administration of nuclease P1 at dosages of 10.1, 101 or 1007 mg TOS/kg bw/day to Sprague-Dawley rats, no adverse effects on any study parameter were observed. The no-observed-adverse-effect level was considered to be 1007 mg TOS/kg bw/day. The results of the genotoxicity studies and subchronic rat study support the safe use in food production of nuclease P1 produced from P. citrinum.

  15. Choline Derivatives Involved in Osmotolerance of Penicillium fellutanum†

    PubMed Central

    Park, Yong-Il; Gander, John E.

    1998-01-01

    Penicillium fellutanum is osmotolerant and xerotolerant when cultured in a low-phosphate medium containing 3 M NaCl. Glycerol and erythritol accumulated in cultures with NaCl concentrations up to 2 M; glycerol was the only detectable polyol in cultures containing 3 M NaCl. In cultures with 3 M NaCl, the intracellular levels of glycine betaine and choline-O-sulfate were 22- and 2.6-fold greater (70 and 46 mM), respectively, than those of cultures without added NaCl. The levels of glycine betaine and glycerol decreased in mycelia transferred from a medium containing 3 M NaCl into a fresh medium without added NaCl. NaCl at 3 M inhibited mycelial mass accumulation; this inhibition was partially corrected by supplementation of cultures with glycine betaine (2 mM) or choline-O-sulfate (10 mM). The presence of exogenous choline chloride (2 mM) in plate cultures protected the cells from stress from 3 M NaCl. The data suggest that glycine betaine and choline-O-sulfate are secondary osmoprotectants which are effective at the point that the cell is incapable of synthesizing more glycerol. PMID:16349488

  16. Insights into Penicillium roqueforti Morphological and Genetic Diversity.

    PubMed

    Gillot, Guillaume; Jany, Jean-Luc; Coton, Monika; Le Floch, Gaétan; Debaets, Stella; Ropars, Jeanne; López-Villavicencio, Manuela; Dupont, Joëlle; Branca, Antoine; Giraud, Tatiana; Coton, Emmanuel

    2015-01-01

    Fungi exhibit substantial morphological and genetic diversity, often associated with cryptic species differing in ecological niches. Penicillium roqueforti is used as a starter culture for blue-veined cheeses, being responsible for their flavor and color, but is also a common spoilage organism in various foods. Different types of blue-veined cheeses are manufactured and consumed worldwide, displaying specific organoleptic properties. These features may be due to the different manufacturing methods and/or to the specific P. roqueforti strains used. Substantial morphological diversity exists within P. roqueforti and, although not taxonomically valid, several technological names have been used for strains on different cheeses (e.g., P. gorgonzolae, P. stilton). A worldwide P. roqueforti collection from 120 individual blue-veined cheeses and 21 other substrates was analyzed here to determine (i) whether P. roqueforti is a complex of cryptic species, by applying the Genealogical Concordance Phylogenetic Species Recognition criterion (GC-PSR), (ii) whether the population structure assessed using microsatellite markers correspond to blue cheese types, and (iii) whether the genetic clusters display different morphologies. GC-PSR multi-locus sequence analyses showed no evidence of cryptic species. The population structure analysis using microsatellites revealed the existence of highly differentiated populations, corresponding to blue cheese types and with contrasted morphologies. This suggests that the population structure has been shaped by different cheese-making processes or that different populations were recruited for different cheese types. Cheese-making fungi thus constitute good models for studying fungal diversification under recent selection.

  17. Microbial Beneficiation of Salem Iron Ore Using Penicillium purpurogenum

    NASA Astrophysics Data System (ADS)

    Mishra, M.; Pradhan, M.; Sukla, L. B.; Mishra, B. K.

    2011-02-01

    High alumina and silica content in the iron ore affects coke rate, reducibility, and productivity in a blast furnace. Iron ore is being beneficiated all around the world to meet the quality requirement of iron and steel industries. Choosing a beneficiation treatment depends on the nature of the gangue present and its association with the ore structure. The advanced physicochemical methods used for the beneficiation of iron ore are generally unfriendly to the environment. Biobeneficiation is considered to be ecofriendly, promising, and revolutionary solutions to these problems. A characterization study of Salem iron ore indicates that the major iron-bearing minerals are hematite, magnetite, and goethite. Samples on average contains (pct) Fe2O3-84.40, Fe (total)-59.02, Al2O3-7.18, and SiO2-7.53. Penicillium purpurogenum (MTCC 7356) was used for the experiment . It removed 35.22 pct alumina and 39.41 pct silica in 30 days in a shake flask at 10 pct pulp density, 308 K (35 °C), and 150 rpm. In a bioreactor experiment at 2 kg scale using the same organism, it removed 23.33 pct alumina and 30.54 pct silica in 30 days at 300 rpm agitation and 2 to 3 l/min aeration. Alumina and silica dissolution follow the shrinking core model for both shake flask and bioreactor experiments.

  18. Talaromyces (Penicillium) marneffei infection in non-HIV-infected patients

    PubMed Central

    Chan, Jasper FW; Lau, Susanna KP; Yuen, Kwok-Yung; Woo, Patrick CY

    2016-01-01

    Talaromyces (Penicillium) marneffei is an important pathogenic thermally dimorphic fungus causing systemic mycosis in Southeast Asia. The clinical significance of T. marneffei became evident when the human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome epidemic arrived in Southeast Asia in 1988. Subsequently, a decline in the incidence of T. marneffei infection among HIV-infected patients was seen in regions with access to highly active antiretroviral therapy and other control measures for HIV. Since the 1990s, an increasing number of T. marneffei infections have been reported among non-HIV-infected patients with impaired cell-mediated immunity. Their comorbidities included primary adult-onset immunodeficiency due to anti-interferon-gamma autoantibodies and secondary immunosuppressive conditions including other autoimmune diseases, solid organ and hematopoietic stem cell transplantations, T-lymphocyte-depleting immunsuppressive drugs and novel anti-cancer targeted therapies such as anti-CD20 monoclonal antibodies and kinase inhibitors. Moreover, improved immunological diagnostics identified more primary immunodeficiency syndromes associated with T. marneffei infection in children. The higher case-fatality rate of T. marneffei infection in non-HIV-infected than HIV-infected patients might be related to delayed diagnosis due to the lack of clinical suspicion. Correction of the underlying immune defects and early use of antifungals are important treatment strategies. Clinicians should be familiar with the changing epidemiology and clinical management of T. marneffei infection among non-HIV-infected patients. PMID:26956447

  19. Insights into Penicillium roqueforti Morphological and Genetic Diversity

    PubMed Central

    Gillot, Guillaume; Jany, Jean-Luc; Coton, Monika; Le Floch, Gaétan; Debaets, Stella; Ropars, Jeanne; López-Villavicencio, Manuela; Dupont, Joëlle; Branca, Antoine; Giraud, Tatiana; Coton, Emmanuel

    2015-01-01

    Fungi exhibit substantial morphological and genetic diversity, often associated with cryptic species differing in ecological niches. Penicillium roqueforti is used as a starter culture for blue-veined cheeses, being responsible for their flavor and color, but is also a common spoilage organism in various foods. Different types of blue-veined cheeses are manufactured and consumed worldwide, displaying specific organoleptic properties. These features may be due to the different manufacturing methods and/or to the specific P. roqueforti strains used. Substantial morphological diversity exists within P. roqueforti and, although not taxonomically valid, several technological names have been used for strains on different cheeses (e.g., P. gorgonzolae, P. stilton). A worldwide P. roqueforti collection from 120 individual blue-veined cheeses and 21 other substrates was analyzed here to determine (i) whether P. roqueforti is a complex of cryptic species, by applying the Genealogical Concordance Phylogenetic Species Recognition criterion (GC-PSR), (ii) whether the population structure assessed using microsatellite markers correspond to blue cheese types, and (iii) whether the genetic clusters display different morphologies. GC-PSR multi-locus sequence analyses showed no evidence of cryptic species. The population structure analysis using microsatellites revealed the existence of highly differentiated populations, corresponding to blue cheese types and with contrasted morphologies. This suggests that the population structure has been shaped by different cheese-making processes or that different populations were recruited for different cheese types. Cheese-making fungi thus constitute good models for studying fungal diversification under recent selection. PMID:26091176

  20. [Penicillium marneffei isolated from a Thai AIDS patient with fungemia].

    PubMed

    Uehara, Masae; Sano, Ayako; Yarita, Kyoko; Kamei, Katsuhiko; Haketa, Makio; Ide, Kyoko; Nagai, Keiko; Takayama, Yoshihiro; Nishimura, Kazuko

    2008-01-01

    Penicillium marneffei was isolated from three blood cultures of a Thai woman with AIDS and then identified as such. The patient, 41 a year-old female from northeast Thailand came to Japan 10 years ago and married a Japanese man. She was reportedly the third patient infected with this fungal species in Japan, and considered to be the first case from whom the causative fungus was successfully cultured, which led to the diagnosis of penicilliosis marneffei. The colony of the isolate, which was cultured on Sabouraud dextrose agar at 25-27 degrees C, was initially white and pannose, gradually turned in color from yellow to yellow-green, and diffused a deep red pigment into the medium. Conidial heads were divergent, and chains of conidia were formed from phialides. Colonies of the isolate, which was cultured on brain-heart infusion agar at 35 degrees C, had a grayish white, membranous yeast-like form with fine plicae and microscopically consisted of short hyphae. Furthermore, 560 bases of the internal transcribed spacer (ITS) region of the ribosomal RNA gene including the 5.8S region (ITS1-5.8S-ITS2) (DDBJ accession number AB298970) were sequenced and allowed an unequivocal species identification.

  1. Isolation and characterization of a novel phytase from Penicillium simplicissimum.

    PubMed

    Tseng, Y H; Fang, T J; Tseng, S M

    2000-01-01

    Eighty-three isolates from different soil samples exhibited the potential for producing active extracellular phytase. The most active fungal isolate with phytase activity was identified as Penicillium simplicissimum. In shaking culture with enrichment medium, the highest extracellular phytase activity of the producing strain was 3.8 U/mL. The crude enzyme filtrate was purified to homogeneity using ultrafiltration. IEC and gel filtration chromatography. The molar mass of the purified enzyme was estimated to be 65 kDa on SDS-PAGE. The saccharide identification with periodic acid-Schiff reagent (PAS) and activity recognition by 1-naphthyl phosphate was all positive. The isoelectric point of the enzyme, as deduced by isoelectric focusing, was pH 5.8, the optimum pH and temperature being pH 4.0 and 55 degrees C, respectively. The purified enzyme revealed broad substrate specificity and was strongly inhibited by Fe2+, Fe3+ and Zn2+; however, no inhibition was found by EDTA and PMSF. Phytase activity was inhibited when 2 mmol/L of dodecasodium phytate was added and the Km for it was determined to be 813 mmol/L.

  2. Measurement of autolysis in submerged batch cultures of Penicillium chrysogenum.

    PubMed

    McNeil, B; Berry, D R; Harvey, L M; Grant, A; White, S

    1998-02-05

    The process of cellular autolysis was studied in an industrial strain of Penicillium chrysogenum by a range of methods, including assessment of biomass decline, NH+4 release, changes in culture apparent viscosity, and by means of a quantitative assessment of changes in micromorphology using a computerized image analysis system. The pattern of total intracellular proteolytic and beta-1, 3-glucanolytic activity in the culture was also examined. The overall aim was to identify a suitable method, or methods, for examining the extent of autolysis in fungal cultures. Autolysis was studied in submerged batch processes, where DOT was maintained above 40% saturation (non-O2-limited), and, under O2-limited conditions. Both N and O2 limitation promoted extensive culture autolysis. Image analysis techniques were perhaps the most sensitive method of assessing the progress of autolysis in the culture. Autolytic regions within some hyphae were apparent even during growth phase, but became much more widespread as the process proceeded. The early stages of autolysis involved continued energy source consumption, increased carbon dioxide evolution rate, degradation of penicillin, and decreased broth filterability. Later stages involved widespread mycelial fragmentation, with some regrowth (cryptic growth) occurring in non-O2-limited cultures. Intracellular proteolytic activity showed two peaks, one during the growth phase, and the other during autolysis. Autolysis was also associated with a distinct peak in beta-1,3-glucanolytic activity, indicating that degradation of cell wall matrix polymers may be occurring during autolysis in this strain of P. chrysogenum.

  3. UV-C light inactivation kinetics of Penicillium expansum on pear surfaces: Influence on physicochemical and sensory quality during storage

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Postharvest quality and storage life of fresh pear are often limited by fungal growth caused by Penicillium expansum. Ultraviolet-C light (UV-C 254 nm) is a promising alternative disinfestation method to reduce fruit spoilage by fungi. In this study, UV-C inactivation kinetic data of Penicillium exp...

  4. Species-specific PCR to describe local-scale distributions of four cryptic species in the Penicillium chrysogenum complex.

    PubMed

    Browne, Alexander G P; Fisher, Matthew C; Henk, Daniel A

    2013-10-01

    Penicillium chrysogenum is a ubiquitous airborne fungus detected in every sampled region of the Earth. Owing to its role in Alexander Fleming's serendipitous discovery of Penicillin in 1928, the fungus has generated widespread scientific interest; however its natural history is not well understood. Research has demonstrated speciation within P. chrysogenum, describing the existence of four cryptic species. To discriminate the four species, we developed protocols for species-specific diagnostic PCR directly from fungal conidia. 430 Penicillium isolates were collected to apply our rapid diagnostic tool and explore the distribution of these fungi across the London Underground rail transport system revealing significant differences between Underground lines. Phylogenetic analysis of multiple type isolates confirms that the 'Fleming species' should be named Penicillium rubens and that divergence of the four 'Chrysogenum complex' fungi occurred about 0.75 million yr ago. Finally, the formal naming of two new species, Penicillium floreyi and Penicillium chainii, is performed.

  5. Penicillium species as a rare isolate in tracheal granulation tissue: a case series

    PubMed Central

    Randhawa, Premjit S; Nouraei, SA Reza; Howard, David J; Sandhu, Gurpreet S; Petrou, Michael A

    2008-01-01

    Introduction Granulation tissue formation is a major problem complicating the treatment of upper airway stenosis. We present two cases of recurrent tracheal granulation tissue colonisation by Penicillium species in patients undergoing laryngotracheal reconstructive surgery for post-intubation tracheal stenosis. We believe that although most Penicillium species do not cause invasive disease they can be a contributory factor to the occurrence of upper airway stenosis. Case presentation A microbiological and mycological study of tracheal granulation tissue in two patients with recurrent laryngotracheal stenosis was carried out. Penicillium species was seen microscopically and cultured from tracheal granulation tissue. Neither patient grew any bacteria known to be associated with airway granulation tissue formation. Amphotericin B, itraconazole, flucytosine voriconazole and caspofungin were highly active against both isolates. Conclusion A search for a fungal cause should form part of the investigation for recurrent tracheal granulation tissue during laryngotracheal reconstruction. PMID:18346276

  6. [Progress in Proteomic Study of the Penicillin Producer---Penicillium Chrysogenum].

    PubMed

    Wang, Shun; Wang, Peihong; Zhang, Nan; Gao, Ruichang

    2015-12-01

    Penicillin is a kind of β-lactam drug which has been applied in the clinical treatment firstly in the world, and it has still been widely used at present. The synthesis and regulation mechanism of Penicillium chrysogenum, which is used to produce penicillin, has been studied quite maturely, but its proteomics research started relatively late and fewer reports were published. This paper reviews the synthesis and application of penicillin, transformation of Penicillium chrysogenum, and the research progress of its proteomics. On this basis, the study highlights the advantages of proteomics in the research of protein expression.

  7. Gibberellins in Penicillium strains: Challenges for endophyte-plant host interactions under salinity stress.

    PubMed

    Leitão, Ana Lúcia; Enguita, Francisco J

    2016-02-01

    The genus Penicillium is one of the most versatile "mycofactories", comprising some species able to produce gibberellins, bioactive compounds that can modulate plant growth and development. Although plants have the ability to synthesize gibberellins, their levels are lower when plants are under salinity stress. It has been recognized that detrimental abiotic conditions, such as saline stress, have negative effects on plants, being the availability of bioactive gibberellins a critical factor for their growth under this conditions. This review summarizes the interplay existing between endophytic Penicillium strains and plant host interactions, with focus on bioactive gibberellins production as a fungal response that allows plants to overcome salinity stress.

  8. Penicillin production by wild isolates of Penicillium chrysogenum in Pakistan.

    PubMed

    Sajjad-Ur-Rahman; Rasool, Muhammad Hidayat; Rafi, Muhammad

    2012-04-01

    The present study was aimed at exploring the native wild isolates of Penicillium chrysogenum series in terms of their penicillin production potential. Apart from the standard medium, the efforts were made to utilize suitable agro-industrial wastes for the maximum yield of penicillin. Two series of P. chrysogenum were isolated from local sources and named as P. chrysogenum series UAF R1 and P. chrysogenum series UAF R2. The native series were found to possess better penicillin production potential than the already reported series of P. chrysogenum. However, P. chrysogenum series UAF R1 was found to be the best candidate for high yield of penicillin starting at 100 hour as compared to P. chrysogenum series UAF R2 which produced the highest yield of penicillin at 150 hours for a shorter period of time. Addition of Corn Steep Liquor (CSL) to the fermentation medium resulted in the production of 1.20g/L penicillin by P. chrysogenum series UAF R1 and P. chrysogenum series UAF R2. The fermentation medium in which Sugar Cane Bagasse (SCB) was replaced with CSL resulted in the highest yield of penicillin (1.92g/L) by both native series of P. chrysogenum. The penicillin production was increased by 62.5% in medium with SCB as compared to that with CSL. The penicillin yield of medium containing lactose and phenyl acetate was higher than that of control medium. Overall results revealed that P. chrysogenum series UAF R1 and P. chrysogenum series UAF R2 may be recommended for better yield of natural penicillin and this efficiency may be further enhanced by utilizing SCB as substrate in the growth medium.

  9. Characterization of an autoinducer of penicillin biosynthesis in Penicillium chrysogenum.

    PubMed

    Martín, Jorge; García-Estrada, Carlos; Rumbero, Angel; Recio, Eliseo; Albillos, Silvia M; Ullán, Ricardo V; Martín, Juan-Francisco

    2011-08-15

    Filamentous fungi produce an impressive variety of secondary metabolites; many of them have important biological activities. The biosynthesis of these secondary metabolites is frequently induced by plant-derived external elicitors and appears to also be regulated by internal inducers, which may work in a way similar to that of bacterial autoinducers. The biosynthesis of penicillin in Penicillium chrysogenum is an excellent model for studying the molecular mechanisms of control of gene expression due to a good knowledge of the biochemistry and molecular genetics of β-lactam antibiotics and to the availability of its genome sequence and proteome. In this work, we first developed a plate bioassay that allows direct testing of inducers of penicillin biosynthesis using single colonies of P. chrysogenum. Using this bioassay, we have found an inducer substance in the conditioned culture broths of P. chrysogenum and Acremonium chrysogenum. No inducing effect was exerted by γ-butyrolactones, jasmonic acid, or the penicillin precursor δ-(L-α-aminoadipyl)-L-cysteinyl-D-valine. The conditioned broth induced penicillin biosynthesis and transcription of the pcbAB, pcbC, and penDE genes when added at inoculation time, but its effect was smaller if added at 12 h and it had no effect when added at 24 h, as shown by Northern analysis and lacZ reporter studies. The inducer molecule was purified and identified by mass spectrometry (MS) and nuclear magnetic resonance (NMR) as 1,3-diaminopropane. Addition of pure 1,3-diaminopropane stimulated the production of penicillin by about 100% compared to results for the control cultures. Genes for the biosynthesis of 1,3-diaminopropane have been identified in the P. chrysogenum genome.

  10. Direct electrochemistry of Penicillium chrysogenum catalase adsorbed on spectroscopic graphite.

    PubMed

    Dimcheva, Nina; Horozova, Elena

    2013-04-01

    The voltammetric studies of Penicillium chrysogenum catalase (PcCAT) adsorbed on spectroscopic graphite, showed direct electron transfer (DET) between its active site and the electrode surface. Analogous tests performed with the commercially available bovine catalase revealed that mammalian enzyme is much less efficient in the DET process. Both catalases were found capable to catalyse the electrooxidation of phenol, but differed in the specifics of catalytic action. At an applied potential of 0.45V the non-linear regression showed the kinetics of the bioelectrochemical oxidation catalysed by the PcCAT obeyed the Hill equation with a binding constant K=0.034±0.002 M(2) (Hill's coefficient n=2.097±0.083, R(2)=0.997), whilst the catalytic action of the bovine catalase was described by the Michaelis-Menten kinetic model with the following parameters: V(max,app)=7.780±0.509 μA, and K(M,app)=0.068±0.070 mol L(-1). The performance of the electrode reaction was affected by the electrode potential, the pH, and temperature. Based on the effect of pH and temperature on the electrode response in presence of phenol a tentative reaction pathway of its bioelectrocatalytic oxidation has been hypothesised. The possible application of these findings in biosensing phenol up to concentration 30 mM at pHs below 7 and in absence of oxidising agents (oxygen or H(2)O(2)) was considered.

  11. Modeling Penicillium expansum resistance to thermal and chlorine treatments.

    PubMed

    Salomão, Beatriz C M; Churey, John J; Aragão, Gláucia M F; Worobo, Randy W

    2009-12-01

    Apples and apple products are excellent substrates for Penicillium expansum to produce patulin. In an attempt to avoid excessive levels of patulin, limiting or reducing P. expansum contamination levels on apples designated for storage in packinghouses and/or during apple juice processing is critical. The aim of this work was (i) to determine the thermal resistance of P. expansum spores in apple juice, comparing the abilities of the Bigelow and Weibull models to describe the survival curves and (ii) to determine the inactivation of P. expansum spores in aqueous chlorine solutions at varying concentrations of chlorine solutions, comparing the abilities of the biphasic and Weibull models to fit the survival curves. The results showed that the Bigelow and Weibull models were similar for describing the heat inactivation data, because the survival curves were almost linear. In this case, the concept of D- and z-values could be used, and the D-values obtained were 10.68, 6.64, 3.32, 1.14, and 0.61 min at 50, 52, 54, 56, and 60 degrees C, respectively, while the z-value was determined to be 7.57 degrees C. For the chlorine treatments, although the biphasic model gave a slightly superior performance, the Weibull model was selected, considering the parsimony principle, because it has fewer parameters than the biphasic model has. In conclusion, the typical pasteurization regimen used for refrigerated apple juice (71 degrees C for 6 s) is capable of achieving a 6-log reduction of P. expansum spores.

  12. Immobilization and Stabilization of Beta-Xylosidases from Penicillium janczewskii.

    PubMed

    Terrasan, César Rafael Fanchini; Romero-Fernández, Maria; Orrego, Alejandro H; Oliveira, Sandro Martins; Pessela, Benevides Costa; Carmona, Eleonora Cano; Guisan, José Manuel

    2016-11-23

    β-Xylosidases are critical for complete degradation of xylan, the second main constituent of plant cell walls. A minor β-xylosidase (BXYL II) from Penicillium janczewskii was purified by ammonium sulfate precipitation (30% saturation) followed by DEAE-Sephadex chromatography in pH 6.5 and elution with KCl. The enzyme presented molecular weight (MW) of 301 kDa estimated by size exclusion chromatography. Optimal activity was observed in pH 3.0 and 70-75 °C, with higher stability in pH 3.0-4.5 and half-lives of 11, 5, and 2 min at 65, 70, and 75 °C, respectively. Inhibition was moderate with Pb(+2) and citrate and total with Cu(+2), Hg(+2), and Co(+2). Partially purified BXYL II and BXYL I (the main β-xylosidase from this fungus) were individually immobilized and stabilized in glyoxyl agarose gels. At 65 °C, immobilized BXYL I and BXYL II presented half-lives of 4.9 and 23.1 h, respectively, therefore being 12.3-fold and 33-fold more stable than their unipuntual CNBr derivatives (reference mimicking soluble enzyme behaviors). During long-term incubation in pH 5.0 at 50 °C, BXYL I and BXYL II glyoxyl derivatives preserved 85 and 35% activity after 25 and 7 days, respectively. Immobilized BXYL I retained 70% activity after 10 reuse cycles of p-nitrophenyl-β-D-xylopyranoside hydrolysis.

  13. Proteome Analysis of the Penicillin Producer Penicillium chrysogenum

    PubMed Central

    Jami, Mohammad-Saeid; Barreiro, Carlos; García-Estrada, Carlos; Martín, Juan-Francisco

    2010-01-01

    Proteomics is a powerful tool to understand the molecular mechanisms causing the production of high penicillin titers by industrial strains of the filamentous fungus Penicillium chrysogenum as the result of strain improvement programs. Penicillin biosynthesis is an excellent model system for many other bioactive microbial metabolites. The recent publication of the P. chrysogenum genome has established the basis to understand the molecular processes underlying penicillin overproduction. We report here the proteome reference map of P. chrysogenum Wisconsin 54-1255 (the genome project reference strain) together with an in-depth study of the changes produced in three different strains of this filamentous fungus during industrial strain improvement. Two-dimensional gel electrophoresis, peptide mass fingerprinting, and tandem mass spectrometry were used for protein identification. Around 1000 spots were visualized by “blue silver” colloidal Coomassie staining in a non-linear pI range from 3 to 10 with high resolution, which allowed the identification of 950 proteins (549 different proteins and isoforms). Comparison among the cytosolic proteomes of the wild-type NRRL 1951, Wisconsin 54-1255 (an improved, moderate penicillin producer), and AS-P-78 (a penicillin high producer) strains indicated that global metabolic reorganizations occurred during the strain improvement program. The main changes observed in the high producer strains were increases of cysteine biosynthesis (a penicillin precursor), enzymes of the pentose phosphate pathway, and stress response proteins together with a reduction in virulence and in the biosynthesis of other secondary metabolites different from penicillin (pigments and isoflavonoids). In the wild-type strain, we identified enzymes to utilize cellulose, sorbitol, and other carbon sources that have been lost in the high penicillin producer strains. Changes in the levels of a few specific proteins correlated well with the improved penicillin

  14. Subcellular localization of the homocitrate synthase in Penicillium chrysogenum.

    PubMed

    Bañuelos, O; Casqueiro, J; Steidl, S; Gutiérrez, S; Brakhage, A; Martín, J F

    2002-01-01

    There are conflicting reports regarding the cellular localization in Saccharomyces cerevisiae and filamentous fungi of homocitrate synthase, the first enzyme in the lysine biosynthetic pathway. The homocitrate synthase (HS) gene (lys1) of Penicillium chrysogenum was disrupted in three transformants (HS(-)) of the Wis 54-1255 pyrG strain. The three mutants named HS1(-), HS2(-) and HS3(-) all lacked homocitrate synthase activity and showed lysine auxotrophy, indicating that there is a single gene for homocitrate synthase in P. chrysogenum. The lys1 ORF was fused in frame to the gene for the green fluorescent protein (GFP) gene of the jellyfish Aequorea victoria. Homocitrate synthase-deficient mutants transformed with a plasmid containing the lys1-GFP fusion recovered prototrophy and showed similar levels of homocitrate synthase activity to the parental strain Wis 54-1255, indicating that the hybrid protein retains the biological function of wild-type homocitrate synthase. Immunoblotting analysis revealed that the HS-GFP fusion protein is maintained intact and does not release the GFP moiety. Fluorescence microscopy analysis of the transformants showed that homocitrate synthase was mainly located in the cytoplasm in P. chrysogenum; in S. cerevisiae the enzyme is targeted to the nucleus. The control nuclear protein StuA was properly targeted to the nucleus when the StuA (targeting domain)-GFP hybrid protein was expressed in P. chrysogenum. The difference in localization of homocitrate synthase between P. chrysogenum and S. cerevisiae suggests that this protein may play a regulatory function, in addition to its catalytic function, in S. cerevisiae but not in P. chrysogenum.

  15. Uptake of phenylacetic acid by two strains of Penicillium chrysogenum.

    PubMed

    Eriksen, S H; Soderblom, T B; Jensen, B; Olsen, J

    1998-11-05

    Uptake of phenylacetic acid, the side-chain precursor of benzylpenicillin, was studied in Penicillium chrysogenum Wisconsin 54-1255 and in a strain yielding high levels of penicillin. In penicillin fermentations with the high-yielding strain, 100% recovery of phenylacetic acid in benzylpenicillin was found, whereas in the Wisconsin strain only 17% of the supplied phenylacetic acid was incorporated into benzylpenicillin while the rest was metabolized. Accumulation of total phenylacetic acid-derived carbon in the cells was nonsaturable in both strains at high external concentrations of phenylacetic acid (250-3500 microM), and in the high-yielding strain at low phenylacetic acid concentrations (2. 8-100 microM), indicating that phenylacetic acid enters the cells by simple diffusion, as concluded earlier for P. chrysogenum by other authors. However, at low external concentrations of phenylacetic acid saturable accumulation appeared in the Wisconsin strain. HPLC-analyses of cell extracts from the Wisconsin strain showed that phenylacetic acid was metabolized immediately after entry into the cells and different [14C]-labeled metabolites were detected in the cells. Up to approximately 50% of the accumulated phenylacetic acid was metabolized during the transport-assay period, the conversion having an impact on the uptake experiments. Nevertheless, accumulation of free unchanged phenylacetic acid in the cells showed saturation kinetics, suggesting the possible involvement of a high-affinity carrier in uptake of phenylacetic acid in P. chrysogenum Wisconsin 54-1255. At high concentrations of phenylacetic acid, contribution to uptake by this carrier is minor in comparison to simple diffusion and therefore, of no importance in the industrial production of penicillin.

  16. Haenamindole and fumiquinazoline analogs from a fungicolous isolate of Penicillium lanosum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two new amino acid-derived compounds, lanosindole (1) and 2'-epi-fumiquinazoline C (2), were isolated from cultures of a fungicolous isolate of Penicillium lanosum (MYC 1813 = NRRL 66231), together with 2'-epi-fumiquinazoline D (3), previously reported only as a product of an in vitro enzymatic step...

  17. Genome sequence of Penicillium solitum RS1, which causes postharvest apple decay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Penicillium species cause postharvest decay, commonly known as blue mold, in pome fruits such as apples and pears. Among the species that cause blue mold, P. expansum is the most virulent and prevalent, while P. solitum is signficantly less virulent. For devising novel strategies to prevent and to r...

  18. Inhibitory effect of selenium against Penicillium expansum and its possible mechanisms of action

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Penicillium expansum is a widely spread fungal pathogen that causes blue mold rot in a variety of fruits. This pathogen not only induces blue mold rot but also produces patulin in affected apple fruit, a secondary metabolite that is toxic to humans and animals. Currently, diseases caused by P. expan...

  19. Ultraviolet-C light inactivation of Penicillium expansum on fruit surfaces

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Understanding the influence of fruit surface morphology on ultraviolet-C (UV-C 254 nm) inactivation of microorganisms is required for designing effective treatment systems. In this study, we analyzed UV-C inactivation of Penicillium expansum that was inoculated onto the surface of organic fruits. Re...

  20. Identification of a QTL for postharvest disease resistance to Penicillium expansum in Malus sieversii

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Blue mold of apple caused by Penicillium expansum is one of the most important postharvest rots of apple fruit. Little attention has been devoted to postharvest disease resistance in apple breeding programs due both to a lack of sources of genetic resistance and to the time required for seedlings t...

  1. Detection of additional genes of the patulin biosynthetic pathway in Penicillium griseofulvum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Genes in the patulin biosynthetic pathway are likely to be arranged in a cluster as has been found for biosynthetic pathways of other mycotoxins. The mycotoxin patulin, common in apples and apple juice, is most often associated with Penicillium expansum. However, of 15 fungal species capable of sy...

  2. Occurrence and phenotypes of pyrimethanil resistance in penicillium expansum from apple in Washington state

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Penicillium expansum is the primary cause of blue mold of apple. Pyrimethanil is a recently registered postharvest fungicide for control of postharvest diseases in apple. To monitor pyrimethanil resistance, 779 isolates of P. expansum were collected from decayed apple fruit in 2010 and 2011 from fiv...

  3. Use of chemosensitization to overcome fludioxonil-resistance in Penicillium expansum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Resistance in two mutant strains of Penicillium expansum to the fungicide fludioxonil was overcome by co-application of natural phenolic chemosensitizing agents that targeted various elements of the oxidative stress-response pathway. Fludioxonil resistance in these strains resulted from cell-linked ...

  4. Growth inhibition of Aspergillus ochraceus ZMPBF 318 and Penicillium expansum ZMPBF 565 by four essential oils.

    PubMed

    Cvek, Domagoj; Markov, Ksenija; Frece, Jadranka; Landeka Dragicević, Tibela; Majica, Matea; Delas, Frane

    2010-06-01

    Fungi produce a large variety of extracellular proteins, organic acids, and other metabolites and can adapt to several environmental conditions. Mycotoxin-producing moulds of the genera Aspergillus and Penicillium are common food contaminants. One of the natural ways to protect food from mould contamination is to use essential oils. In this study, we evaluated the effect of essential oils of cinnamon, lavender, rosemary, and sage at 1 % (v/v) concentration in yeast media inoculated with spores (final concentration 106 mL-1 media) of Aspergillus ochraceus ZMPBF 318 and Penicillium expansum ZMPBF 565, alone or in combination, on fungal biomass. Cinnamon showed the best inhibitory effect (100 %). Lavender oil best inhibited the growth of Aspergillus ochraceus (nearly 100 %), and was less successful with Penicillium expansum (having dropped to 57 % on day 28). With cultivation time the inhibitory effect of sage and rosemary oil grew for Aspergillus ochraceus and dropped for Penicillium expansum.These results suggest that fungi can be controlled with essential oils, especially with cinnamon oil.

  5. A Rapid Assay to Detect Toxigenic Penicillium spp. Contamination in Wine and Musts

    PubMed Central

    Sanzani, Simona Marianna; Miazzi, Monica Marilena; di Rienzo, Valentina; Fanelli, Valentina; Gambacorta, Giuseppe; Taurino, Maria Rosaria; Montemurro, Cinzia

    2016-01-01

    Wine and fermenting musts are grape products widely consumed worldwide. Since the presence of mycotoxin-producing fungi may greatly compromise their quality characteristics and safety, there is an increasing need for relatively rapid “user friendly” quantitative assays to detect fungal contamination both in grapes delivered to wineries and in final products. Although other fungi are most frequently involved in grape deterioration, secondary infections by Penicillium spp. are quite common, especially in cool areas with high humidity and in wines obtained by partially dried grapes. In this work, a single-tube nested real-time PCR approach—successfully applied to hazelnut and peanut allergen detection—was tested for the first time to trace Penicillium spp. in musts and wines. The method consisted of two sets of primers specifically designed to target the β-tubulin gene, to be simultaneously applied with the aim of lowering the detection limit of conventional real-time PCR. The assay was able to detect up to 1 fg of Penicillium DNA. As confirmation, patulin content of representative samples was determined. Most of analyzed wines/musts returned contaminated results at >50 ppb and a 76% accordance with molecular assay was observed. Although further large-scale trials are needed, these results encourage the use of the newly developed method in the pre-screening of fresh and processed grapes for the presence of Penicillium DNA before the evaluation of related toxins. PMID:27509524

  6. The biochemical basis of pathogenicity and host-specificity of Penicillium digitatum on citrus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In this work, we report that volatiles emitted from wounded citrus peel play a major role in host recognition by Penicillium digitatum. Volatiles of various citrus cultivars had a pronounced stimulatory effect on germination and germ tube elongation of green mold pathogen. When exposed to volatile...

  7. DOSE-DEPENDENT ALLERGIC RESPONSES TO AN EXTRACT OF PENICILLIUM CHRYSOGENUM IN BALB/MICE

    EPA Science Inventory

    Indoor mold has been associated with the development of allergic asthma. Penicillium chrysogenum, a common indoor mold, is known to have several allergens and can induce allergic responses in a mouse model of allergic penicilliosis. Our hypothesis is that soluble components of ...

  8. DOSE-DEPENDENT ALLERGIC RESPONSES TO AN EXTRACT OF PENICILLIUM CHRYSOGENUM IN BAL/C MICE

    EPA Science Inventory

    Indoor mold has been associated with the development of allergic asthma. Penicillium chrysogenum, a common indoor mold, is known to have several allergens and can induce allergic responses in a mouse model of allergic penicilliosis. Our hypothesis is that soluble components of ...

  9. AN EXTRACT OF PENICILLIUM CHRYSOGENUM INDUCES DOSE-DEPENDENT ALLERGIC ASTHMA RESPONSES IN MICE

    EPA Science Inventory

    Rationale: Penicillium chrysogenum, a common indoor mold, is known to have several allergens and can induce allergic responses in a mouse model of allergic penicilliosis. Our hypothesis is that soluble components of P. chrysogenum (PCE) can dose-dependently induce responses typ...

  10. Host ranges of Penicillium species causing blue mold of bulb crops in Washington State and Idaho

    Technology Transfer Automated Retrieval System (TEKTRAN)

    First reported from the Pacific Northwest (PNW) of U.S.A. as causal agents of blue mold on edible and/or ornamental bulbs are Penicillium albocoremium (from Tulipa sp.; pathogenic on Allium sativum, A. cepa, A. stipitatum, Iris hollandica and Tulipa sp.), P. crustosum (from Narcissus; pathogenic on ...

  11. [Isolation of imazalil and prochloraz resistant Penicillium variable strains from a commercial citrus warehouse].

    PubMed

    Díaz, M A; Vila, R; Hernández, E

    1987-10-01

    "In vitro" sensitivity of two strains of Penicillium variabile to different concentrations of Imazalil and Prochloraz has been studied. Both strains were isolated from a citrus packing-house in which Imazalil was used. These strains were resistant to both compounds showing a ED50 higher than 100 ppm.

  12. First report of Penicillium crustosum causing blue mold on stored apple fruit in Serbia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Penicillium crustosum Thom causes blue mold on pome fruits and is also regularly found on cheese, nuts and soil. The fungus produces an array of mycotoxins that impact human health, including penitrem A, roquefortine C, terrestric acid, and cyclopenol. In January and February 2013, decayed apples, ‘...

  13. First report of Penicillium carneum causing blue mold on stored apples in the United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Blue mold decay occurs during long term storage of apples and is predominantly caused by Penicillium expansum Link. Apples harvested in 2010 were stored in controlled atmosphere at a commercial Pennsylvania apple packing and storage facility, and were examined for occurrence of decay in May 2011. ...

  14. Characterization of blue mold Penicillium species isolated from stored fruits using multiple highly conserved loci

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Penicillium is a large genus of common molds with over 400 described species; however, identification of individual species is difficult, including for those species that cause postharvest rots. In this study, blue rot fungi from stored apples and pears were isolated from a variety of hosts, locatio...

  15. First report of Penicillium expansum isolates resistant to pyrimethanil from stored apple fruit in Pennsylvania

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Apples in the United States are stored in low temperature controlled atmosphere for 9–12 months and are susceptible to decay by blue mold. Penicillium spp. cause significant economic losses worldwide and produce mycotoxins that contaminate processed apple products. Blue mold is managed by a combinat...

  16. ASSESSMENT OF IMMUNE RESPONSES TO PENICILLIUM CHRYSOGENUM AND CHARACTERIZATION OF ITS ALLERGENS

    EPA Science Inventory

    Assessment of immune responses to Penicillium chrysogenum and characterization of its allergens

    Yongjoo Chung1, Michael E Viana2, Lisa B Copeland3, and MaryJane K Selgrade3, Marsha D W Ward3. 1 UNC, SPH, Chapel Hill, NC, 2NCSU, CVM, Raleigh, NC, 3US EPA, ORD, NHEERL, RTP,...

  17. Performance of fogged disinfectants to inactivate conida of Penicillium digitatum within citrus degreening rooms

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fogging with formaldehyde of citrus packinghouses when the fruit are absent is a practice to control conidia of Penicillium digitatum (Pers.) Sacc., cause of citrus green mold. Replacements for formaldehyde in these facilities are needed because of worker and environmental health issues. To evaluate...

  18. A new polyketide, penicillolide from the marine-derived fungus Penicillium sacculum.

    PubMed

    Liu, Tao; Zhang, Songya; Li, Zhanlin; Wang, Yu; Chen, Zaixing; Bai, Jiao; Tian, Li; Pei, Yuehu; Hua, Huiming

    2016-01-01

    A new polyketide, penicillolide (1) was isolated from the fermentation broth of the marine-derived fungus Penicillium sacculum GT-308. Compound 1 is a polyketide with a unique carbon skeleton. The structure of this compound was established via extensive spectroscopic analyses including 1D-, 2D-NMR, and HRESI-MS.

  19. Occurrence of fludioxonil resistance in penicillium digitatum from citrus in California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Penicillium digitatum is the causal agent of green mold, the most important postharvest disease of citrus (Citrus spp.). Fludioxonil is marketed as either a solo product or in combination with azoxystrobin for control of green mold and other postharvest diseases. Baseline sensitivity to fludioxonil ...

  20. Endophytic synthesis of silver chloride nanoparticles from Penicillium sp. of Calophyllum apetalum

    NASA Astrophysics Data System (ADS)

    Chandrappa, C. P.; Govindappa, M.; Chandrasekar, N.; Sarkar, Sonia; Ooha, Sepuri; Channabasava, R.

    2016-06-01

    In the present study, Penicillium species extract isolated from Calophyllum apetalum was used for the synthesis of silver nanoparticles and it was confirmed by changing the color of the silver nitrate UV-Vis spectrum. The synthesized nanoparticles have been characterized by biophysical techniques such as scanning electron microscopy and x-ray diffraction.

  1. Novel decaturin alkaloids from the marine-derived fungus Penicillium oxalicum.

    PubMed

    Wang, Pei-le; Lib, Dan-yi; Xie, Lei-rui; Wu, Xin; Hua, Hui-ming; Li, Zhan-lin

    2013-10-01

    From the mycelia of Penicillium oxalicum two new compounds, decaturins E (1) and F (2), have been isolated, along with four known analogues, decaturin A (3), decaturin C (4), decaturin D (5), and oxalicine B (6). The structures were determined by HR-ESI-MS and 1D and 2D NMR analysis.

  2. Detection of VM55599 and preparaherquamide from Aspergillus japonicus and Penicillium fellutanum: biosynthetic implications.

    PubMed

    Ding, Yousong; Gruschow, Sabine; Greshock, Thomas J; Finefield, Jennifer M; Sherman, David H; Williams, Robert M

    2008-09-01

    The secondary metabolites VM55599 (4) and preparaherquamide (5) have been identified by LC-MS(n) analysis as natural metabolites in cultures of Penicillium fellutanum, whereas preparaherquamide has been identified only in cultures of Aspergillus japonicus. In accord with a previous proposal, the identification of both metabolites, which have a diastereomeric relationship, provides indirect support for a unified biosynthetic scheme.

  3. Production of bioactive compounds based on phylogeny in the genus Penicillium preserved at NBRC.

    PubMed

    Nakashima, Takuji; Mayuzumi, Shinzo; Inaba, Shigeki; Park, Ju-Young; Anzai, Kozue; Suzuki, Rieko; Kuwahara, Natsumi; Utsumi, Noriko; Yokoyama, Fumie; Sato, Hajime; Okane, Izumi; Tsurumi, Yasuhisa; Ando, Katsuhiko

    2008-11-01

    Penicillium strains (n=394) preserved at NBRC (the NITE Biological Resource Center) were compared as to groupings (11 species-clusters) based on phylogeny and the production of bioactive compounds. The strains in two clusters, of which P. chrysogenum and P. citrinum are representative, showed higher rates of positive strains with multi-biological activities.

  4. Two New Records of Penicillium Associated with Blue Moldy Bulbs of Lily in Korea

    PubMed Central

    Kim, Won Ki; Park, Myung Soo; Hahm, Soo-Sang

    2006-01-01

    Two new records of Penicillium from blue moldy bulbs of lily are reported in Korea. The Korean isolates of P. albocoremium (Frisvad) Frisvad and P. tulipae Overy and Frisvad were phylogenetically identical to the reference species based on DNA sequence of the β-tubulin gene. P. albocoremium and P. tulipae are described and illustrated. PMID:24039494

  5. Protein profiling of the dimorphic, pathogenic fungus, Penicillium marneffei

    PubMed Central

    Chandler, Julie M; Treece, Erin R; Trenary, Heather R; Brenneman, Jessica L; Flickner, Tressa J; Frommelt, Jonathan L; Oo, Zaw M; Patterson, Megan M; Rundle, William T; Valle, Olga V; Kim, Thomas D; Walker, Gary R; Cooper, Chester R

    2008-01-01

    Background Penicillium marneffei is a pathogenic fungus that afflicts immunocompromised individuals having lived or traveled in Southeast Asia. This species is unique in that it is the only dimorphic member of the genus. Dimorphism results from a process, termed phase transition, which is regulated by temperature of incubation. At room temperature, the fungus grows filamentously (mould phase), but at body temperature (37°C), a uninucleate yeast form develops that reproduces by fission. Formation of the yeast phase appears to be a requisite for pathogenicity. To date, no genes have been identified in P. marneffei that strictly induce mould-to-yeast phase conversion. In an effort to help identify potential gene products associated with morphogenesis, protein profiles were generated from the yeast and mould phases of P. marneffei. Results Whole cell proteins from the early stages of mould and yeast development in P. marneffei were resolved by two-dimensional gel electrophoresis. Selected proteins were recovered and sequenced by capillary-liquid chromatography-nanospray tandem mass spectrometry. Putative identifications were derived by searching available databases for homologous fungal sequences. Proteins found common to both mould and yeast phases included the signal transduction proteins cyclophilin and a RACK1-like ortholog, as well as those related to general metabolism, energy production, and protection from oxygen radicals. Many of the mould-specific proteins identified possessed similar functions. By comparison, proteins exhibiting increased expression during development of the parasitic yeast phase comprised those involved in heat-shock responses, general metabolism, and cell-wall biosynthesis, as well as a small GTPase that regulates nuclear membrane transport and mitotic processes in fungi. The cognate gene encoding the latter protein, designated RanA, was subsequently cloned and characterized. The P. marneffei RanA protein sequence, which contained the

  6. Native pears of Sardinia affect Penicillium expansum pathogenesis.

    PubMed

    Cubaiu, L; Azara, E; Ladu, G; Venditti, T; D'Hallewin, G

    2013-01-01

    Penicillium expansum causes blue mould rot, a serious post-harvest disease of pome fruits and is the main producer of the mycotoxin patulin. The occurrence of natural resistance against different hostpathogens, has been evidenced in some pear accessions of the Sardinian germoplasm. The aim of this research was to correlate P. expansum growth and patulin production on these indigenous pear accessions. In vitro and in vivo experiments were carried out with seven accessions ('Sarmentina', 'Vacchesa', 'De Puleu', 'De su Duca', 'Natalina', 'Oliena', 'Laconi 5') belonging to the CNR-ISPA ex situ collection and one national control cultivar ('Abate'). A wild type P. expansum from our collection was isolated from blue mould-decayed Sardinian pear fruit and selected for its aggressiveness and patulin production. The in vivo assay was carried out using 5 x 2 cm (Ø x thickness) sterilized fruit discs wounded and inoculated by a 10(5)UFC/mL concentration of P. expansum. Fruit discs were incubated at 23 degrees C for 7 days before analysis. The in vitro experiments, aimed at monitoring over time P. expansum mycelial growth and patulin accumulation, were performed with a standard medium (PDA) and a pear puree Agar Medium (PAM). Petri dishes with PDA and PAM were inoculated centrally with P. expansum conidia (10(5)UFC/ml) and then incubated at 23 degrees C for 7 days. Mycelial growth on Sardinian PAMs was inhibited in comparison to 'Abate' PAM and PDA. In particular, the accessions 'Sarmentina' and 'Vacchesa' showed the maximum inhibitory activity both in vitro and in vivo. Patulin production was detected by high-pressure liquid chromatography-mass spectrometry. The mycotoxin concentration in Sardinian PAMs was lower than that detected in PDA medium, pointing out a positive correlation between fungal growth inhibition and patulin production. The lowest concentration of patulin was found in 'Sarmentina' PAM. Based on these findings, some of Sardinian pear accessions seems to

  7. Enhancement of Penicillium echinulatum glycoside hydrolase enzyme complex.

    PubMed

    dos Santos Costa, Patrícia; Büchli, Fernanda; Robl, Diogo; Delabona, Priscila da Silva; Rabelo, Sarita Candida; Pradella, José Geraldo da Cruz

    2016-05-01

    The enhancement of enzyme complex produced by Penicillium echinulatum grown in several culture media components (bagasse sugarcane pretreated by various methods, soybean meal, wheat bran, sucrose, and yeast extract) was studied to increment FPase, xylanase, pectinase, and β-glucosidase enzyme activities. The present results indicated that culture media composed with 10 g/L of the various bagasse pretreatment methods did not have any substantial influence with respect to the FPase, xylanase, and β-glucosidase attained maximum values of, respectively, 2.68 FPU/mL, 2.04, and 115.4 IU/mL. On the other hand, proposed culture media to enhance β-glucosidase production composed of 10 g/L steam-exploded bagasse supplemented with soybean flour 5.0 g/L, yeast extract 1.0 g/L, and sucrose 10.0 g/L attained, respectively, 3.19 FPU/mL and 3.06 IU/mL while xylanase was maintained at the same level. The proteomes obtained from the optimized culture media for enhanced FPase, xylanase, pectinase, and β-glucosidase production were analyzed using mass spectrometry and a panel of GH enzyme activities against 16 different substrates. Culture medium designed to enhance β-glucosidase activity achieved higher enzymatic activities values (13 measured activities), compared to the culture media for FPase/pectinase (9 measured activities) and xylanase (7 measured activities), when tested against the 16 substrates. Mass spectrometry analyses of secretome showed a consistent result and the greatest number of spectral counts of Cazy family enzymes was found in designed β-glucosidase culture medium, followed by FPase/pectinase and xylanase. Most of the Cazy identified protein was cellobiohydrolase (GH6 and GH7), endoglucanase (GH5), and endo-1,4-β-xylanase (GH10). Enzymatic hydrolysis of hydrothermally pretreated sugarcane bagasse performed with β-glucosidase enhanced cocktail achieved 51.4 % glucose yield with 10 % w/v insoluble solids at enzyme load of 15 FPU/g material. Collectively the

  8. Functional studies of ATP sulfurylase from Penicillium chrysogenum

    SciTech Connect

    Seubert, P.A.

    1985-01-01

    ATP sulfurylase from Penicillium chrysogenum has a specific activity (V/sub max/) of 6-7 units x mg protein/sup -1/ determined with the physiological substrates of MgATP and SO/sub 4//sup 2 -/ and assayed by (A) initial velocity measurements with APS kinase and inorganic pyrophosphatase present and (B) analysis of nonlinear reaction progress curves. The fact both assays give the same results show the intrinsic activity of ATP sulfurylase is much higher than previously reported. In initial velocity dead-end inhibition studies, the sulfate analog S/sub 2/O/sub 3//sup 2 -/ is a competitive inhibitor of SO/sub 42/..sqrt.. and a noncompetitive inhibitor of MgATP. Monovalent oxyanions such as NO/sub 3//sup -/, ClO/sub 3//sup -/, ClO/sub 4//sup -/, and FSO/sub 3//sup -/ behave as uncompetitive inhibitors of MgATP and thus seem not to be true sulfate analogs. The reverse reaction was assayed by the pyrophosphate dependent release of /sup 35/SO/sub 4//sup 2 -/ from AP/sup 35/S. Product inhibition by MgATP or SO/sub 4//sup 2 -/ is competitive with APS and mixed-type with PP/sub i/. Imidodiphosphate can serve as an alternative substrate for PP/sub i/. ATP sulfurylase binds (but does not hydrolyze) APS. A Scatchard plot of the APS binding is nonlinear, suggesting at least two types of sites. The cumulative results are qualitatively consistent with the random addition of MgATP and SO/sub 4//sup 2 -/ and the ordered release of first MgPP/sub i/ then APS, with APS release being partially rate limiting. Certain quantitative discrepancies suggest either an unknown variable (e.g. enzyme concentration) complicates the analysis or, in light of binding studies that the actual mechanism is more complicated (e.g. alternating sites) than any of the conventional models examined.

  9. Fungal flora of the digestive tract of 5 species of triatomines vectors of Trypanosoma cruzi, Chagas 1909.

    PubMed

    Moraes, A M; Junqueira, A C; Costa, G L; Celano, V; Oliveira, P C; Coura, J R

    2001-01-01

    A study of the mycobiota in the digestive tract of 5 important species of triatomines, Triatoma brasiliensis, T infestans, T. sordida, T. pseudomaculata and T. vitticeps, was made. The digestive tracts of 164 adults and 535 nymphs of those triatomines were studied and 393 fungal strains were isolated. The genera with the greatest number of species were Penicillium (19 species), Aspergillus (17 species) and Acremonium (5 species) and the most frequent species, in decreasing order, were Penicillium corylophilum, Aspergillus niger, Penicillium felluttanum, Cladosporium herbarum, Penicillium waksmanii, Aspergillus awamori and Paecilomyces variotii. Among the isolated fungi, we found species that are recognized as entomopathogenic and pathogenic for humans and animals.

  10. DESTRUCTION OF ASPERGILLUS VERSICOLOR, PENICILLIUM CRYSOGENUM, STACHYBOTRYS CHARTARUM, AND CLADOSPORIUM CLADOSPORIDES SPORES USING CHEMICAL OXIDATION TREATMENT PROCESS

    EPA Science Inventory

    The survival of aqueous suspensions of Penicillium chrysogenum, Stachybotrys chartarum, Aspergillus versicolor, and Cladosporium cladosporioides spores was evaluated using various combinations of hydrogen peroxide and iron (II) as catalyst. Spores were suspended in water and trea...

  11. Discrimination of Penicillium isolates by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry fingerprinting.

    PubMed

    Hettick, Justin M; Green, Brett J; Buskirk, Amanda D; Kashon, Michael L; Slaven, James E; Janotka, Erika; Blachere, Francoise M; Schmechel, Detlef; Beezhold, Donald H

    2008-08-01

    Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used to generate highly reproducible mass spectral 'fingerprints' for twelve Penicillium species. Prior to MALDI-TOF MS analysis, eight replicate cultures of each Penicillium species were subjected to three one-minute bead-beating cycles in an acetonitrile/trifluoroacetic acid solvent. The mass spectra contained abundant peaks in the range of m/z 5000-20 000, and allowed unambiguous discrimination between species. In addition, a biomarker common to all Penicillium mass spectra was observed at m/z 13 900. Discriminant analysis using the MALDI-TOF MS data yielded classification error rates of 0% (i.e. 100% correct identification), indicating that MALDI-TOF MS data may be a useful diagnostic tool for the objective identification of Penicillium species of environmental and clinical importance.

  12. Phylogeny of Penicillium and the segregation of Trichocomaceae into three families

    PubMed Central

    Houbraken, J.; Samson, R.A.

    2011-01-01

    Species of Trichocomaceae occur commonly and are important to both industry and medicine. They are associated with food spoilage and mycotoxin production and can occur in the indoor environment, causing health hazards by the formation of β-glucans, mycotoxins and surface proteins. Some species are opportunistic pathogens, while others are exploited in biotechnology for the production of enzymes, antibiotics and other products. Penicillium belongs phylogenetically to Trichocomaceae and more than 250 species are currently accepted in this genus. In this study, we investigated the relationship of Penicillium to other genera of Trichocomaceae and studied in detail the phylogeny of the genus itself. In order to study these relationships, partial RPB1, RPB2 (RNA polymerase II genes), Tsr1 (putative ribosome biogenesis protein) and Cct8 (putative chaperonin complex component TCP-1) gene sequences were obtained. The Trichocomaceae are divided in three separate families: Aspergillaceae, Thermoascaceae and Trichocomaceae. The Aspergillaceae are characterised by the formation flask-shaped or cylindrical phialides, asci produced inside cleistothecia or surrounded by Hülle cells and mainly ascospores with a furrow or slit, while the Trichocomaceae are defined by the formation of lanceolate phialides, asci borne within a tuft or layer of loose hyphae and ascospores lacking a slit. Thermoascus and Paecilomyces, both members of Thermoascaceae, also form ascospores lacking a furrow or slit, but are differentiated from Trichocomaceae by the production of asci from croziers and their thermotolerant or thermophilic nature. Phylogenetic analysis shows that Penicillium is polyphyletic. The genus is re-defined and a monophyletic genus for both anamorphs and teleomorphs is created (Penicillium sensu stricto). The genera Thysanophora, Eupenicillium, Chromocleista, Hemicarpenteles and Torulomyces belong in Penicillium s. str. and new combinations for the species belonging to these genera

  13. The effect of Penicillium fungi on plant growth and phosphorus mobilization in neutral to alkaline soils from southern Australia.

    PubMed

    Wakelin, S A; Gupta, V V S R; Harvey, P R; Ryder, M H

    2007-01-01

    The phosphate solubilizing fungi Penicillium radicum, Penicillium bilaiae (strain RS7B-SD1), and an unidentified Penicillium sp. designated strain KC6-W2 were tested for their ability to increase the growth and phosphorus (P) nutrition of wheat, medic, and lentil in three soils of neutral to alkaline pH reaction. The strongest plant growth promoting (PGP) strain was Penicillium sp. KC6-W2, which stimulated significant increases in shoot growth and dry mass in seven of the nine experiments conducted. Levels of PGP by Penicillium sp. KC6-W2 ranged from 6.6% to 19% and were associated with increased uptake of P to the shoot. The PGP properties of Penicillium sp. KC6-W2 were evident on each of the three different plant species and soil types, a level of reliability not observed in other strains tested. Inoculation of seed with P. radicum increased lentil growth by 5.5% (P < 0.05) in soil from Tarlee but did not affect plant growth in the eight other experiments. Inoculation of plant seed with P. bilaiae RS7B-SD1 resulted in significant PGP in two of the nine experiments conducted. However, when significant, stimulation of PGP by P. bilaiae RS7B-SD1 was strong and resulted in increases in medic dry matter (19%) and lentil shoot dry matter (15%). A soil microcosm experiment investigated the effect of Penicillium fungi on cycling of soil P. Penicillium bilaiae RS7B-SD1 was the only fungus to significantly increase HCO3-extractable P (23% increase; P < 0.05). Production of phosphatase enzymes was not associated with increased HCO3-extractable P. Addition of carbon in the form of ryegrass seed significantly increased microbial respiration and movement of P to the microbial biomass (P < 0.05), but these parameters were irrespective of Penicillium treatment. This work has established the potential for use of Penicillium inoculants to increase plant growth on alkaline soils in Australia. The role of Penicillium fungi in plant P uptake and soil P cycling requires further

  14. Survival of Penicillium spp. conidia during deep-frying and baking steps of frozen chicken nuggets processing.

    PubMed

    Wigmann, Évelin Francine; Moreira, Rafael Chelala; Alvarenga, Verônica Ortiz; Sant'Ana, Anderson S; Copetti, Marina Venturini

    2016-05-01

    This study aimed at determining whether Penicillium spp. strains could survive through the heat treatment applied during the processing of frozen chicken nuggets. Firstly, it was found that the conidia of Penicillium were not able to survive the heat shock in phosphate buffer at pH 7.2 in thermal death tubes (TDT) at 80 °C/30 min. Subsequently, each Penicillium strain was inoculated in frozen chicken nuggets, which were subjected to the following treatments: i) only deep frying (frying oil at 195-200 °C), ii) only baking (120-130 °C until the internal temperature reached 70 °C) and iii) deep frying followed by baking (frying oil temperature of 195-200 °C and baking temperature of 120-130 °C, until the internal temperature reached 70 °C). The results indicated that Penicillium polonicum NGT 23/12, Penicillium commune NGT 16/12, Penicillium solitum NGT 30/12 and Penicillium crustosum NGT 51/12 were able to survive after the combined treatment (deep frying followed by baking) when inoculated in chicken nuggets. P. polonicum NGT 23/12 was the most resistant strain to the combined treatment (deep frying and baking), as its population was reduced by 3 log cycles CFU/g, when the internal temperature reached 78 °C after 10 min and 30 s of baking. The present data show that if Penicillium spp. is present in high numbers in raw materials, such as breading flours, it will survive the thermal processing applied during chicken nuggets production.

  15. An evaluation of the proteolytic and lipolytic potential of Penicillium spp. isolated from traditional Greek sausages in submerged fermentation.

    PubMed

    Papagianni, Maria

    2014-01-01

    A number of novel Penicillium strains belonging to Penicillium nalgiovense, Penicillium solitum, Penicillium commune, Penicillium olsonii, and Penicillium oxalicum species, isolated from the surface of traditional Greek sausages, were evaluated for their proteolytic and lipolytic potential in a solid substrate first and next in submerged fermentations, using complex media. Extracellular proteolytic activity was assessed at acid, neutral, and alkaline pH, while the lipolytic activity was assessed using olive oil, the short-chain triacylglycerol tributyrin, and the long-chain triolein, as substrates. The study revealed that although closely related, the tested strains produce enzymes of distinct specificities. P. nalgiovense PNA9 produced the highest alkaline proteolytic activity (13.2 unit (U)/ml) and the highest lipolytic activity with tributyrin (92 U/ml). Comparisons with known sources show that proteases and/or lipases can be secreted effectively by some Penicillia (P. nalgiovense PNA4, PNA7, and PNA9 and P. solitum PSO1), and further investigations on their properties and characteristics would be promising.

  16. Multilocus sequence identification of Penicillium species in cork bark during plank preparation for the manufacture of stoppers.

    PubMed

    Serra, Rita; Peterson, Stephen; Venâncio, Armando

    2008-04-01

    Despite several studies reporting Penicillium as one of the most frequent fungal genera in cork planks, the isolates were rarely identified to species level. We conducted a detailed study to identify Penicillium species from the field to the factory environment prior to and after boiling the cork planks. A total of 84 samples were analyzed. Of the 486 Penicillium isolates phenotypically identified, 32 representative or unusual strains were selected for identification by multilocus DNA sequence type. Cork proved to be a rich source of Penicillium biodiversity. A total of 30 taxa were recognized from cork including rarely seen species and 6 phylogenetically unique groups. Spores of some species lodged deep in cork can survive the boiling process. P. glabrum, P. glandicola and P. toxicarium, species with high CFU numbers in the field, are still frequently present in cork after boiling. Other species are killed by the boiling treatment and replaced by Penicillium species originating from the factory environment. Species known to contribute to cork taint were isolated at all stages. Good manufacturing practices are necessary at all stages in the preparation of cork planks to minimize the load of Penicillium species that produce cork taint.

  17. The small molecular mass antifungal protein of Penicillium chrysogenum--a mechanism of action oriented review.

    PubMed

    Hegedus, Nikoletta; Leiter, Eva; Kovács, Barbara; Tomori, Valéria; Kwon, Nak-Jung; Emri, Tamás; Marx, Florentine; Batta, Gyula; Csernoch, László; Haas, Hubertus; Yu, Jae-Hyuk; Pócsi, István

    2011-12-01

    The β-lactam producing filamentous fungus Penicillium chrysogenum secretes a 6.25 kDa small molecular mass antifungal protein, PAF, which has a highly stable, compact 3D structure and is effective against a wide spectrum of plant and zoo pathogenic fungi. Its precise physiological functions and mode of action need to be elucidated before considering possible biomedical, agricultural or food technological applications. According to some more recent experimental data, PAF plays an important role in the fine-tuning of conidiogenesis in Penicillium chrysogenum. PAF triggers apoptotic cell death in sensitive fungi, and cell death signaling may be transmitted through two-component systems, heterotrimeric G protein coupled signal transduction and regulatory networks as well as via alteration of the Ca(2+) -homeostasis of the cells. Possible biotechnological applications of PAF are also outlined in the review.

  18. Biosorption of binary mixtures of copper and cobalt by Penicillium brevicompactum.

    PubMed

    Tsekova, Kolishka; Ianis, Maria; Dencheva, Vera; Ganeva, Sonya

    2007-01-01

    This work reports on a study of the biosorption of copper and cobalt, both singly and in combination (in equimolar concentrations), by the resting cells of Penicillium brevicompactum. Equilibrium batch sorption studies were carried out at 30 degrees C and pH 5.0 for a contact time of 1 hour to guarantee that equilibrium was reached. The equilibrium data were analyzed using the Langmuir and Freundlich isotherms. The adsorption of binary mixtures of heavy metal solutions on the fungal biomass was found to be of competitive type where the adsorption capacity for any single metal decreased in the presence of the other. The cobalt ions showed a higher affinity for Penicillium brevicompactum than the copper ions.

  19. Purification of Candida guilliermondii and Pichia ohmeri killer toxin as an active agent against Penicillium expansum.

    PubMed

    Coelho, Alexandre Rodrigo; Tachi, Masahico; Pagnocca, Fernando Carlos; Nobrega, Gisele Maria Andrade; Hoffmann, Fernando Leite; Harada, Ken-Ichi; Hirooka, Elisa Yoko

    2009-01-01

    An antifungal assay with cell-free culture supernatant of Pichia ohmeri 158 and Candida guilliermondii P3 was tested against Penicillium expansum strain #2 at 25 degrees C by measuring hyphal length and percentage conidia germination. C. guilliermondii was more effective against P. expansum conidia germination (58.15% inhibition), while P. ohmeri showed higher inhibition of mycelial growth (66.17%), indicating a probable mechanism associated with killer activity. This killer toxin (molecular mass <3 kDa) was partially purified by normal phase HPLC, using TSKgel Amide-80 analytical and preparative columns. Compared with crude extract, the killer toxin eluted from the post analytical column significantly inhibited P. expansum:% inhibition rose from 42.16 to 90.93% (C. guilliermondii) and 39.32 to 91.12% (P. ohmeri) (p < 0.05). The one-step purification process was adequate in isolating killer toxin from culture supernatant and also increased anti-Penicillium activity.

  20. Yaequinolones J1 and J2, novel insecticidal antibiotics from Penicillium sp. FKI-2140.

    PubMed

    Uchida, Ryuji; Imasato, Rie; Shiomi, Kazuro; Tomoda, Hiroshi; Omura, Satoshi

    2005-12-08

    [chemical reaction: see text]. Two novel insecticidal antibiotics with a p-methoxyphenylquinolinone skeleton fused with a pyran ring, yaequinolones J1 (1) and J2 (2), have been isolated from Penicillium sp. FKI-2140, and structures were elucidated by spectroscopic studies including various NMR experiments. The relative stereochemistries were assigned by NOE experiments. Yaequinolones J1 and J2 showed toxicity against Artemia salina (brine shrimp) with the MIC value of 6.25 microg/mL.

  1. Abundance of airborne Penicillium CFU in relation to urbanization in Mexico City.

    PubMed

    Rosas, I; Calderón, C; Ulloa, M; Lacey, J

    1993-08-01

    Air was sampled simultaneously at three localities in Mexico City differing in urbanization index and air pollution level on 22 days during a period covering both dry and rainy seasons. An Andersen two-stage microbial sampler was used for 15 min at 28 liters min-1 to isolate culturable fungi on malt extract agar. After exposure, plates were incubated at 25 degrees C for 48 to 72 h before colonies were counted and identified to give concentrations of total fungal spores and of Penicillium spp., expressed as CFU per cubic meter of air. Total fungi numbered 91 to 602 CFU m-3 in Tlalpan Borough (southern area), 40 to 264 CFU m-3 in Cuauhtémoc Borough (downtown), and 26 to 495 CFU m-3 in Gustavo A. Madero Borough (northern area). Although Penicillium spp. were the second most frequently isolated fungal genus, concentrations were small, with a maximum of only 133 CFU m-3. Twice as many colonies were isolated in the southern area, with an urbanization index of 0.25 (arithmetic mean, 41 CFU m-3), as at other sampling stations with greater urbanization indices (arithmetic means, 19 and 20 CFU m-3). In the downtown area, with an urbanization index of 1.0, Penicillium spp. were more numerous than any other genus and formed 25% of the total fungal count compared with 14 and 17% in the other areas. Concentrations of airborne Penicillium spp. did not differ significantly between rainy and dry seasons.(ABSTRACT TRUNCATED AT 250 WORDS)

  2. Effect of dissolved carbon dioxide on penicillin fermentations: mycelial growth and penicillin production. [Penicillium chrysogenum

    SciTech Connect

    Ho, C.S.; Smith, M.D.

    1986-01-01

    The effect of dissolved carbon dioxide on the specific growth rate and the penicillin production rate of Penicillium chrysogenum was examined experimentally. The dissolved carbon dioxide was found to inhibit the specific growth rate and the penicillin production rate when the aerated submerged penicillin fermentation was exposed to influent gases of 12.6 and 20% carbon dioxide, respectively. Upon exposure to influent gases of 3 and 5% carbon dioxide, no pronounced metabolic inhibition was noted.

  3. Abundance of airborne Penicillium CFU in relation to urbanization in Mexico City.

    PubMed Central

    Rosas, I; Calderón, C; Ulloa, M; Lacey, J

    1993-01-01

    Air was sampled simultaneously at three localities in Mexico City differing in urbanization index and air pollution level on 22 days during a period covering both dry and rainy seasons. An Andersen two-stage microbial sampler was used for 15 min at 28 liters min-1 to isolate culturable fungi on malt extract agar. After exposure, plates were incubated at 25 degrees C for 48 to 72 h before colonies were counted and identified to give concentrations of total fungal spores and of Penicillium spp., expressed as CFU per cubic meter of air. Total fungi numbered 91 to 602 CFU m-3 in Tlalpan Borough (southern area), 40 to 264 CFU m-3 in Cuauhtémoc Borough (downtown), and 26 to 495 CFU m-3 in Gustavo A. Madero Borough (northern area). Although Penicillium spp. were the second most frequently isolated fungal genus, concentrations were small, with a maximum of only 133 CFU m-3. Twice as many colonies were isolated in the southern area, with an urbanization index of 0.25 (arithmetic mean, 41 CFU m-3), as at other sampling stations with greater urbanization indices (arithmetic means, 19 and 20 CFU m-3). In the downtown area, with an urbanization index of 1.0, Penicillium spp. were more numerous than any other genus and formed 25% of the total fungal count compared with 14 and 17% in the other areas. Concentrations of airborne Penicillium spp. did not differ significantly between rainy and dry seasons.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8368852

  4. Tanzawaic acids I–L: Four new polyketides from Penicillium sp. IBWF104-06

    PubMed Central

    Sandjo, Louis P; Thines, Eckhard

    2014-01-01

    Summary Four new polyketides have been identified in culture filtrates of the fungal strain Penicillium sp. IBWF104-06 isolated from a soil sample. They are structurally based on the same trans-decalinpentanoic acid skeleton as tanzawaic acids A–H. One of the new compounds was found to inhibit the conidial germination in the rice blast fungus Magnaporthe oryzae at concentrations of 25 μg/mL. PMID:24605144

  5. Effects of salts and temperatures on post-irradiation growth of Penicillium exposed to ultraviolet

    SciTech Connect

    Valdez, R.; Siegel, B.Z.; Siegel, S.M.

    1981-01-01

    The growth of Penicillium notatum colonies after uv irradiation of dried mycelium or spores was studied in relation to post-irradiation temperature and salt environment. Dried mycelium and spores behaved differently with respect to sensitivity to temperature, salts and uv, especially the latter. Threshold inhibitory doses for spores were modified markedly either at 4 C or in magnesium and calcium chlorides. It is suggested that these temperature and salt effects are related to prevention of photochemical membrane damage.

  6. Mycobiota and toxigenic Penicillium species on two Spanish dry-cured ham manufacturing plants.

    PubMed

    Alapont, C; López-Mendoza, M C; Gil, J V; Martínez-Culebras, P V

    2014-01-01

    The present study reports the natural mycobiota occurring in dry-cured hams, and in particular on the incidence of mycotoxin-producing fungi. A total of 338 fungal colonies were isolated from three stages of production, these being the post-salting, ripening and aging stages in two manufacturing plants. The results show that fungi were more frequently isolated from the aging stage and that the predominant filamentous fungal genus isolated was Penicillium. Seventy-four of the 338 fungal strains were selected for identification at the species level by using morphological criteria and internal transcribed spacers sequencing. Of the 74 fungal strains, 59 were Penicillium strains. Sixteen Penicillium species were identified, with P. commune (24 strains) and P. chrysogenum (13 strains) being the most abundant. The potential ability to produce cyclopiazonic acid (CPA) and ochratoxin A (OTA) was studied by isolating the culture followed by HPLC analysis of these mycotoxins in the culture extracts. The results indicated that 25 (33.7%) of the 74 fungal strains produced CPA. Worth noting is the high percentage of CPA-producing strains of P. commune (66.6%) of which some strains were highly toxigenic. P. polonicum strains were also highly toxigenic. With respect to OTA-producing fungi, a low percentage of fungal strains (9.5%) were able to produce OTA at moderate levels. OTA-producing fungi belonged to different Penicillium species including P. chrysogenum, P. commune, P. polonicum and P. verrucosum. These results indicate that there is a possible risk factor posed by CPA and OTA contamination of dry-cured hams.

  7. [Biosynthesis of biologically active low-molecular weight compounds by fungi of the genus Penicillium (review)].

    PubMed

    Kozlovskii, A G; Antipova, T V; Zhelifonova, V P

    2015-01-01

    The recent data on exometabolite biosynthesis in fungi of the genus Penicillium is summarized. The study of creative species, as well as those isolated from extreme ecotopes, resulted in the identification of a number of novel, biologically active compounds. Alkaloid biosynthesis has been shown to begin on.the first day of fungus cultivation and to proceed throughout the cultivation period. Idiophase kinetics was observed for the biosynthesis of polyketide metabolites. The mechanisms of regulation of biosynthesis of promising bioactive compounds are discussed.

  8. Impact of the Penicillium chrysogenum genome on industrial production of metabolites.

    PubMed

    van den Berg, Marco Alexander

    2011-10-01

    The genome sequence of Penicillium chrysogenum has initiated a range of fundamental studies, deciphering the genetic secrets of the industrial penicillin producer. More than 60 years of classical strain improvement has resulted in major but delicate rebalancing of the intracellular metabolism leading to the impressive penicillin titres of the current production strains. Several leads for further improvement are being followed up, including the use of P. chrysogenum as a cell factory for other products than β-lactam antibiotics.

  9. Proteomics shows new faces for the old penicillin producer Penicillium chrysogenum.

    PubMed

    Barreiro, Carlos; Martín, Juan F; García-Estrada, Carlos

    2012-01-01

    Fungi comprise a vast group of microorganisms including the Ascomycota (majority of all described fungi), the Basidiomycota (mushrooms or higher fungi), and the Zygomycota and Chytridiomycota (basal or lower fungi) that produce industrially interesting secondary metabolites, such as β-lactam antibiotics. These compounds are one of the most commonly prescribed drugs world-wide. Since Fleming's initial discovery of Penicillium notatum 80 years ago, the role of Penicillium as an antimicrobial source became patent. After the isolation of Penicillium chrysogenum NRRL 1951 six decades ago, classical mutagenesis and screening programs led to the development of industrial strains with increased productivity (at least three orders of magnitude). The new "omics" era has provided the key to understand the underlying mechanisms of the industrial strain improvement process. The review of different proteomics methods applied to P. chrysogenum has revealed that industrial modification of this microorganism was a consequence of a careful rebalancing of several metabolic pathways. In addition, the secretome analysis of P. chrysogenum has opened the door to new industrial applications for this versatile filamentous fungus.

  10. Genome, Transcriptome, and Functional Analyses of Penicillium expansum Provide New Insights Into Secondary Metabolism and Pathogenicity.

    PubMed

    Ballester, Ana-Rosa; Marcet-Houben, Marina; Levin, Elena; Sela, Noa; Selma-Lázaro, Cristina; Carmona, Lourdes; Wisniewski, Michael; Droby, Samir; González-Candelas, Luis; Gabaldón, Toni

    2015-03-01

    The relationship between secondary metabolism and infection in pathogenic fungi has remained largely elusive. The genus Penicillium comprises a group of plant pathogens with varying host specificities and with the ability to produce a wide array of secondary metabolites. The genomes of three Penicillium expansum strains, the main postharvest pathogen of pome fruit, and one Pencillium italicum strain, a postharvest pathogen of citrus fruit, were sequenced and compared with 24 other fungal species. A genomic analysis of gene clusters responsible for the production of secondary metabolites was performed. Putative virulence factors in P. expansum were identified by means of a transcriptomic analysis of apple fruits during the course of infection. Despite a major genome contraction, P. expansum is the Penicillium species with the largest potential for the production of secondary metabolites. Results using knockout mutants clearly demonstrated that neither patulin nor citrinin are required by P. expansum to successfully infect apples. Li et al. ( MPMI-12-14-0398-FI ) reported similar results and conclusions in their recently accepted paper.

  11. Proteomics Shows New Faces for the Old Penicillin Producer Penicillium chrysogenum

    PubMed Central

    Barreiro, Carlos; Martín, Juan F.; García-Estrada, Carlos

    2012-01-01

    Fungi comprise a vast group of microorganisms including the Ascomycota (majority of all described fungi), the Basidiomycota (mushrooms or higher fungi), and the Zygomycota and Chytridiomycota (basal or lower fungi) that produce industrially interesting secondary metabolites, such as β-lactam antibiotics. These compounds are one of the most commonly prescribed drugs world-wide. Since Fleming's initial discovery of Penicillium notatum 80 years ago, the role of Penicillium as an antimicrobial source became patent. After the isolation of Penicillium chrysogenum NRRL 1951 six decades ago, classical mutagenesis and screening programs led to the development of industrial strains with increased productivity (at least three orders of magnitude). The new “omics” era has provided the key to understand the underlying mechanisms of the industrial strain improvement process. The review of different proteomics methods applied to P. chrysogenum has revealed that industrial modification of this microorganism was a consequence of a careful rebalancing of several metabolic pathways. In addition, the secretome analysis of P. chrysogenum has opened the door to new industrial applications for this versatile filamentous fungus. PMID:22318718

  12. A rapid knockdown effect of Penicillium citrinum for control of the mosquito Culex quinquefasciatus in Thailand.

    PubMed

    Maketon, Monchan; Amnuaykanjanasin, Alongkorn; Kaysorngup, Achirayar

    2014-02-01

    Twenty local isolates of entomopathogenic fungi were determined for control of the larvae and adults of Culex quinquefasciatus. In a laboratory experiment, a Penicillium sp. CM-010 caused 100% mortality of third-instar larvae within 2 h using a conidial suspension of 1 × 10⁶ conidia ml⁻¹. Its LC₅₀ was 3 × 10⁵ conidia ml⁻¹, and the lethal time (LT₅₀) was 1.06 h. Cloning and sequencing of its internal transcribed spacer region indicated that this Penicillium species is Penicillium citrinum (100% identity in 434 bp). Mortality of the adult was highest with Aspergillus flavus CM-011 followed with Metarhizium anisopliae CKM-048 from 1 × 10⁹ conidia ml⁻¹. P. citrinum CM-010 at 1 × 10⁶ conidia ml⁻¹ killed 100% larvae within 2 h while Bacillus thuringiensis var. israelensis at 5 ITU ml⁻¹ required 24 h. This P. citrinum CM-010 also greatly reduced survival of C. quinquefasciatus larvae in an unreplicated field test. Light and transmission electron micrographs showed that the fungal conidia were ingested by the larvae and deposited in the gut. The metabolite patulin was produced by P. citrinum CM-010 instead of citrinin.

  13. Cytotoxicity and mycotoxin production of shellfish-derived Penicillium spp., a risk for shellfish consumers.

    PubMed

    Geiger, M; Guitton, Y; Vansteelandt, M; Kerzaon, I; Blanchet, E; Robiou du Pont, T; Frisvad, J C; Hess, P; Pouchus, Y F; Grovel, O

    2013-11-01

    In order to assess the putative toxigenic risk associated with the presence of fungal strains in shellfish-farming areas, Penicillium strains were isolated from bivalve molluscs and from the surrounding environment, and the influence of the sample origin on the cytotoxicity of the extracts was evaluated. Extracts obtained from shellfish-derived Penicillia exhibited higher cytotoxicity than the others. Ten of these strains were grown on various media including a medium based on mussel extract (Mytilus edulis), mussel flesh-based medium (MES), to study the influence of the mussel flesh on the production of cytotoxic compounds. The MES host-derived medium was created substituting the yeast extract of YES medium by an aqueous extract of mussel tissues, with other constituent identical to YES medium. When shellfish-derived strains of fungi were grown on MES medium, extracts were found to be more cytotoxic than on the YES medium for some of the strains. HPLC-UV/DAD-MS/MS dereplication of extracts from Penicillium marinum and P. restrictum strains grown on MES medium showed the enhancement of the production of some cytotoxic compounds. The mycotoxin patulin was detected in some P. antarcticum extracts, and its presence seemed to be related to their cytotoxicity. Thus, the enhancement of the toxicity of extracts obtained from shellfish-derived Penicillium strains grown on a host-derived medium, and the production of metabolites such as patulin suggests that a survey of mycotoxins in edible shellfish should be considered.

  14. Genomic Characterization Reveals Insights Into Patulin Biosynthesis and Pathogenicity in Penicillium Species.

    PubMed

    Li, Boqiang; Zong, Yuanyuan; Du, Zhenglin; Chen, Yong; Zhang, Zhanquan; Qin, Guozheng; Zhao, Wenming; Tian, Shiping

    2015-06-01

    Penicillium species are fungal pathogens that infect crop plants worldwide. P. expansum differs from P. italicum and P. digitatum, all major postharvest pathogens of pome and citrus, in that the former is able to produce the mycotoxin patulin and has a broader host range. The molecular basis of host-specificity of fungal pathogens has now become the focus of recent research. The present report provides the whole genome sequence of P. expansum (33.52 Mb) and P. italicum (28.99 Mb) and identifies differences in genome structure, important pathogenic characters, and secondary metabolite (SM) gene clusters in Penicillium species. We identified a total of 55 gene clusters potentially related to secondary metabolism, including a cluster of 15 genes (named PePatA to PePatO), that may be involved in patulin biosynthesis in P. expansum. Functional studies confirmed that PePatL and PePatK play crucial roles in the biosynthesis of patulin and that patulin production is not related to virulence of P. expansum. Collectively, P. expansum contains more pathogenic genes and SM gene clusters, in particular, an intact patulin cluster, than P. italicum or P. digitatum. These findings provide important information relevant to understanding the molecular network of patulin biosynthesis and mechanisms of host-specificity in Penicillium species.

  15. Evaluation of secretome of highly efficient lignocellulolytic Penicillium sp. Dal 5 isolated from rhizosphere of conifers.

    PubMed

    Rai, Rohit; Kaur, Baljit; Singh, Surender; Di Falco, Macros; Tsang, Adrian; Chadha, B S

    2016-09-01

    Penicillium sp. (Dal 5) isolated from rhizosphere of conifers from Dalhousie (Himachal Pradesh, India) was found to be an efficient cellulolytic strain. The culture under shake flask on CWR (cellulose, wheat bran and rice straw) medium produced appreciably higher levels of endoglucanase (35.69U/ml), β-glucosidase (4.20U/ml), cellobiohydrolase (2.86U/ml), FPase (1.2U/ml) and xylanase (115U/ml) compared to other Penicillium strains reported in literature. The mass spectroscopy analysis of Penicillium sp. Dal 5 secretome identified 108 proteins constituting an array of CAZymes including glycosyl hydrolases (GH) belonging to 24 different families, polysaccharide lyases (PL), carbohydrate esterases (CE), lytic polysaccharide mono-oxygenases (LPMO) in addition to swollenin and a variety of carbohydrate binding modules (CBM) indicating an elaborate genetic potential of this strain for hydrolysis of lignocellulosics. Further, the culture extract was evaluated for hydrolysis of alkali treated rice straw, wheat straw, bagasse and corn cob at 10% substrate loading rate.

  16. Prospects for fungus identification using CO1 DNA barcodes, with Penicillium as a test case.

    PubMed

    Seifert, Keith A; Samson, Robert A; Dewaard, Jeremy R; Houbraken, Jos; Lévesque, C André; Moncalvo, Jean-Marc; Louis-Seize, Gerry; Hebert, Paul D N

    2007-03-06

    DNA barcoding systems employ a short, standardized gene region to identify species. A 648-bp segment of mitochondrial cytochrome c oxidase 1 (CO1) is the core barcode region for animals, but its utility has not been tested in fungi. This study began with an examination of patterns of sequence divergences in this gene region for 38 fungal taxa with full CO1 sequences. Because these results suggested that CO1 could be effective in species recognition, we designed primers for a 545-bp fragment of CO1 and generated sequences for multiple strains from 58 species of Penicillium subgenus Penicillium and 12 allied species. Despite the frequent literature reports of introns in fungal mitochondrial genomes, we detected introns in only 2 of 370 Penicillium strains. Representatives from 38 of 58 species formed cohesive assemblages with distinct CO1 sequences, and all cases of sequence sharing involved known species complexes. CO1 sequence divergences averaged 0.06% within species, less than for internal transcribed spacer nrDNA or beta-tubulin sequences (BenA). CO1 divergences between species averaged 5.6%, comparable to internal transcribed spacer, but less than values for BenA (14.4%). Although the latter gene delivered higher taxonomic resolution, the amplification and alignment of CO1 was simpler. The development of a barcoding system for fungi that shares a common gene target with other kingdoms would be a significant advance.

  17. Biotransformation of 2,4-dinitroanisole by a fungal Penicillium sp.

    PubMed

    Schroer, Hunter W; Langenfeld, Kathryn L; Li, Xueshu; Lehmler, Hans-Joachim; Just, Craig L

    2017-02-01

    Insensitive munitions explosives are new formulations that are less prone to unintended detonation compared to traditional explosives. While these formulations have safety benefits, the individual constituents, such as 2,4-dinitroanisole (DNAN), have an unknown ecosystem fate with potentially toxic impacts to flora and fauna exposed to DNAN and/or its metabolites. Fungi may be useful in remediation and have been shown to degrade traditional nitroaromatic explosives, such as 2,4,6-trinitrotoluene and 2,4-dinitrotoluene, that are structurally similar to DNAN. In this study, a fungal Penicillium sp., isolated from willow trees and designated strain KH1, was shown to degrade DNAN in solution within 14 days. Stable-isotope labeled DNAN and an untargeted metabolomics approach were used to discover 13 novel transformation products. Penicillium sp. KH1 produced DNAN metabolites resulting from ortho- and para-nitroreduction, demethylation, acetylation, hydroxylation, malonylation, and sulfation. Incubations with intermediate metabolites such as 2-amino-4-nitroanisole and 4-amino-2-nitroanisole as the primary substrates confirmed putative metabolite isomerism and pathways. No ring-cleavage products were observed, consistent with other reports that mineralization of DNAN is an uncommon metabolic outcome. The production of metabolites with unknown persistence and toxicity suggests further study will be needed to implement remediation with Penicillium sp. KH1. To our knowledge, this is the first report on the biotransformation of DNAN by a fungus.

  18. Effects of different culture media on biodegradation of triclosan by Rhodotorula mucilaginosa and Penicillium sp.

    PubMed

    Ertit Taştan, Burcu; Özdemir, Caner; Tekinay, Turgay

    Triclosan is an antimicrobial agent and a persistent pollutant. The biodegradation of triclosan is dependent on many variables including the biodegradation organism and the environmental conditions. Here, we evaluated the triclosan degradation potential of two fungi strains, Rhodotorula mucilaginosa and Penicillium sp., and the rate of its turnover to 2,4-dichlorophenol (2,4-DCP). Both of these strains showed less susceptibility to triclosan when grown in minimal salt medium. In order to further evaluate the effects of environmental conditions on triclosan degradation, three different culture conditions including original thermal power plant wastewater, T6 nutrimedia and ammonium mineral salts medium were used. The maximum triclosan degradation yield was 48% for R. mucilaginosa and 82% for Penicillium sp. at 2.7 mg/L triclosan concentration. Biodegradation experiments revealed that Penicillium sp. was more tolerant to triclosan. Scanning electron microscopy micrographs also showed the morphological changes of fungus when cells were treated with triclosan. Overall, these fungi strains could be used as effective microorganisms in active uptake (degradation) and passive uptake (sorption) of triclosan and their efficiency can be increased by optimizing the culture conditions.

  19. Use of GFP-tagged strains of Penicillium digitatum and Penicillium expansum to study host-pathogen interactions in oranges and apples.

    PubMed

    Buron-Moles, G; López-Pérez, M; González-Candelas, L; Viñas, I; Teixidó, N; Usall, J; Torres, R

    2012-11-15

    Penicillium digitatum and Penicillium expansum are responsible for green and blue molds in citrus and pome fruits, respectively, which result in major monetary losses worldwide. In order to study their infection process in fruits, we successfully introduced a green fluorescent protein (GFP) encoding gene into wild type P. digitatum and P. expansum isolates, using Agrobacterium tumefaciens-mediated transformation (ATMT), with hygromycin B resistance as the selectable marker. To our knowledge, this is the first report describing the transformation of these two important postharvest pathogens with GFP and the use of transformed strains to study compatible and non-host pathogen interactions. Transformation did not affect the pathogenicity or the ecophysiology of either species compared to their respective wild type strains. The GFP-tagged strains were used for in situ analysis of compatible and non-host pathogen interactions on oranges and apples. Knowledge of the infection process of apples and oranges by these pathogens will facilitate the design of novel strategies to control these postharvest diseases and the use of the GFP-tagged strains will help to determine the response of P. digitatum and P. expansum on/in plant surface and tissues to different postharvest treatments.

  20. The effect of locust bean gum (LBG)-based edible coatings carrying biocontrol yeasts against Penicillium digitatum and Penicillium italicum causal agents of postharvest decay of mandarin fruit.

    PubMed

    Parafati, Lucia; Vitale, Alessandro; Restuccia, Cristina; Cirvilleri, Gabriella

    2016-09-01

    Strains belonging to Wickerhamomyces anomalus, Metschnikowia pulcherrima and Aureobasidium pullulans species were tested in vitro as biocontrol agents (BCAs) against the post-harvest pathogenic molds Penicillium digitatum and Penicillium italicum. Moreover, studies aimed at screening the antifungal activity of selected yeast strains in vivo conditions against P. digitatum and P. italicum, and investigated the efficacy of a polysaccharidic matrix, locust bean gum (LBG), enriched with the tested BCAs, in controlling postharvest decays in artificially inoculated mandarins. The population dynamics of BCAs on wounds and the magnitude of peroxidase (POD) and superoxide dismutase (SOD) in fruit tissues were also investigated after treatments of mandarins with antagonistic yeasts. W. anomalus BS91, M. pulcherrima MPR3 and A. pullulans PI1 provided excellent control of postharvest decays caused by P. digitatum and P. italicum on mandarins, both when the yeasts were used alone and in combination with LBG, which enhanced the yeast cell viability over time. Finally, the increased activity of POD and lower decrease in SOD activity in response to BCAs application in mandarin fruits confirmed their involvement in the biocontrol mechanism.

  1. Inhibition of Penicillium digitatum and Penicillium italicum by hydroxypropyl methylcellulose-lipid edible composite films containing food additives with antifungal properties.

    PubMed

    Valencia-Chamorro, Silvia A; Palou, Lluís; del Río, Miguel A; Pérez-Gago, María B

    2008-12-10

    New hydroxypropyl methylcellulose (HPMC)-lipid edible composite films containing low-toxicity chemicals with antifungal properties were developed. Tested chemicals were mainly salts of organic acids, salts of parabens, and mineral salts, classified as food additives or generally recognized as safe (GRAS) compounds. Selected films containing food preservatives were used for in vitro evaluation (disk diameter test) of their antifungal activity against Penicillium digitatum (PD) and Penicillium italicum (PI), the most important postharvest pathogens of fresh citrus fruit. Mechanical properties and oxygen (OP) and water vapor permeabilities (WVP) of selected films were also determined. Film disks containing parabens and their mixtures inhibited PD and PI to a higher extent than the other chemicals tested. Among all organic acid salts tested, potassium sorbate (PS) and sodium benzoate (SB) were the most effective salts in controlling both PD and PI. The use of mixtures of parabens or organic acid salts did not provide an additive or synergistic effect for mold inhibition when compared to the use of single chemicals. Barrier and mechanical properties of films were affected by the addition of food preservatives. Results showed that HPMC-lipid films containing an appropriate food additive should promise as potential commercial antifungal edible coatings for fresh citrus fruit.

  2. Modelling the effect of temperature, pH, water activity, and organic acids on the germination time of Penicillium camemberti and Penicillium roqueforti conidia.

    PubMed

    Kalai, Safaa; Anzala, Lexane; Bensoussan, Maurice; Dantigny, Philippe

    2017-01-02

    In this study, the influence of environmental factors on the germination time of Penicillium camemberti and Penicillium roqueforti conidia was evaluated. To do so, the effects of i/temperature, pH, water activity, and ii/organic acids were determined using models based on i/cardinal values, and ii/minimum inhibitory concentration (MIC) respectively. Cardinal values for germination of conidia were not observed to be species dependent. Minimum temperatures were estimated to be below the freezing point, with an optimum of 26.9°C, and a maximum of 33.5°C. For both species, minimal and optimal aw values were found to be 0.83 and 0.99, respectively, while for pH these values corresponded to 2.9, and 5.6. MIC values could not be determined for lactic acid because conidia of both species germinated in up to 1M concentrations, the highest concentration tested. At pH5.6, P. camemberti (MIC=0.197M) was more sensitive to propionic acid than P. roqueforti (MIC=0.796M).

  3. The draft genome sequence of the ascomycete fungus Penicillium subrubescens reveals a highly enriched content of plant biomass related CAZymes compared to related fungi.

    PubMed

    Peng, Mao; Dilokpimol, Adiphol; Mäkelä, Miia R; Hildén, Kristiina; Bervoets, Sander; Riley, Robert; Grigoriev, Igor V; Hainaut, Matthieu; Henrissat, Bernard; de Vries, Ronald P; Granchi, Zoraide

    2017-03-20

    Here we report the genome sequence of the ascomycete saprobic fungus Penicillium subrubescens FBCC1632/CBS132785 isolated from a Jerusalem artichoke field in Finland. The 39.75Mb genome containing 14,188 gene models is highly similar for that reported for other Penicillium species, but contains a significantly higher number of putative carbohydrate active enzyme (CAZyme) encoding genes.

  4. Examination of the taxonomic position of Penicillium strains used in blue cheese production based on the partial sequence of β-tubulin.

    PubMed

    Ogawa, Yoshio; Hirose, Dai; Akiyama, Ayano; Ichinoe, Masakatsu

    2014-01-01

    Penicillium roqueforti is a well known starter used for blue cheese production. Two closely related species, P. carneum and P. paneum, were previously classified as varieties of P. roqueforti. Penicillium roqueforti does not produce patulin, a mycotoxin harmful for human health, whereas both P. carneum and P. paneum actively produce this toxin. From the viewpoint of food safety, it is thus important to confirm that P. carneum and P. paneum are not used for cheese production. In the present study, the taxonomic position of Penicillium strains used for blue cheese production was examined on the basis of the partial sequence of β-tubulin. Twenty-eight Penicillium strains isolated from blue cheeses were investigated. All the examined strains belonged to P. roqueforti. Therefore, the Penicillium strains used for production of the blue cheese samples examined here do not negatively impact on human health.

  5. Identification, characterization, and molecular cloning of a novel hyaluronidase, a member of glycosyl hydrolase family 16, from Penicillium spp.

    PubMed

    Bakke, Mikio; Kamei, Jun-ichi; Obata, Akio

    2011-01-03

    Hyaluronidase (HAase) activity was detected in the culture supernatants of Penicillium purpurogenum and Penicillium funiculosum. The HAase from Penicillium spp. (HAase-P) was a hyaluronate 4-glycanohydrolase, which catalyzed the endolytic hydrolysis of the β-1,4 glycosidic linkage, as do vertebrate HAases. The gene encoding HAase-P was cloned and expressed in Escherichia coli. According to homology analyses of the deduced amino acid sequences, HAase-P is not classified into any of the known HAase groups, but belongs to glycoside hydrolase family 16, which includes endo-β-1,3(4)-glucanase. Regarding the substrate specificities, no chondroitinase and glucanase activities were detected. Judging from homology analyses and enzymatic properties, HAase-P seems to be a new type of HAase.

  6. Evaluation of food additives and low-toxicity compounds as alternative chemicals for the control of Penicillium digitatum and Penicillium italicum on citrus fruit.

    PubMed

    Palou, Lluís; Usall, Josep; Smilanick, Joseph L; Aguilar, Maria-José; Viñas, Inmaculada

    2002-05-01

    The effectiveness of low-toxicity chemicals as possible alternatives to synthetic fungicides for the control of post-harvest green and blue moulds of citrus was evaluated. A preliminary selection of chemicals, mostly common food additives, was made through in vivo primary screenings with oranges artificially inoculated with Penicillium digitatum or P italicum. Selected compounds and mixtures were tested as heated solutions in small-scale trials. Immersion of artificially inoculated oranges or lemons for 120 s in solutions at 40.6 degrees C and natural pH of potassium sorbate (0.2 M), sodium benzoate (0.2 M) or mixtures (0.1 + 0.1 M) of potassium sorbate with sodium benzoate, sodium propionate or sodium acetate were the most effective organic acid salts tested and reduced green mould by 70-80% after 7 days of storage at 20 degrees C. The mixtures did not significantly enhance the effectiveness of potassium sorbate or sodium benzoate alone. These solutions were as effective as sodium carbonate or calcium polysulphide treatments and, in general, they were more effective on lemons than on oranges. Satisfactory control of green and blue moulds was obtained by dipping oranges for 150 s in solutions of sodium molybdate (24.2 mM) or ammonium molybdate (1.0 mM) at 48 or 53 degrees C, but not at 20 degrees C. At 53 degrees C, however, the effectiveness of hot water was not enhanced by either molybdate. Molybdenum salts at higher concentrations were phytotoxic and stained the fruit. At non-phytotoxic concentrations, the effectiveness of these solutions was more influenced by temperature than by concentration. In general, the inhibitory effects of all compounds tested were not fungicidal but fungistatic and not very persistent. In conclusion, potassium sorbate, sodium benzoate and ammonium molybdate, among the wide range of chemicals tested, were superior for the control of post-harvest Penicillium decay of citrus fruit.

  7. Molecular characterization of the PR-toxin gene cluster in Penicillium roqueforti and Penicillium chrysogenum: cross talk of secondary metabolite pathways.

    PubMed

    Hidalgo, Pedro I; Ullán, Ricardo V; Albillos, Silvia M; Montero, Olimpio; Fernández-Bodega, María Ángeles; García-Estrada, Carlos; Fernández-Aguado, Marta; Martín, Juan-Francisco

    2014-01-01

    The PR-toxin is a potent mycotoxin produced by Penicillium roqueforti in moulded grains and grass silages and may contaminate blue-veined cheese. The PR-toxin derives from the 15 carbon atoms sesquiterpene aristolochene formed by the aristolochene synthase (encoded by ari1). We have cloned and sequenced a four gene cluster that includes the ari1 gene from P. roqueforti. Gene silencing of each of the four genes (named prx1 to prx4) resulted in a reduction of 65-75% in the production of PR-toxin indicating that the four genes encode enzymes involved in PR-toxin biosynthesis. Interestingly the four silenced mutants overproduce large amounts of mycophenolic acid, an antitumor compound formed by an unrelated pathway suggesting a cross-talk of PR-toxin and mycophenolic acid production. An eleven gene cluster that includes the above mentioned four prx genes and a 14-TMS drug/H(+) antiporter was found in the genome of Penicillium chrysogenum. This eleven gene cluster has been reported to be very poorly expressed in a transcriptomic study of P. chrysogenum genes under conditions of penicillin production (strongly aerated cultures). We found that this apparently silent gene cluster is able to produce PR-toxin in P. chrysogenum under static culture conditions on hydrated rice medium. Noteworthily, the production of PR-toxin was 2.6-fold higher in P. chrysogenum npe10, a strain deleted in the 56.8kb amplifiable region containing the pen gene cluster, than in the parental strain Wisconsin 54-1255 providing another example of cross-talk between secondary metabolite pathways in this fungus. A detailed PR-toxin biosynthesis pathway is proposed based on all available evidence.

  8. Functional characterization of the penicillin biosynthetic gene cluster of Penicillium chrysogenum Wisconsin54-1255.

    PubMed

    van den Berg, Marco A; Westerlaken, Ilja; Leeflang, Chris; Kerkman, Richard; Bovenberg, Roel A L

    2007-09-01

    Industrial strain improvement via classical mutagenesis is a black box approach. In an attempt to learn from and understand the mutations introduced, we cloned and characterized the amplified region of industrial penicillin production strains. Upon amplification of this region Penicillium chrysogenum is capable of producing an increased amount of antibiotics, as was previously reported [Barredo, J.L., Diez, B., Alvarez, E., Martín, J.F., 1989a. Large amplification of a 35-kb DNA fragment carrying two penicillin biosynthetic genes in high yielding strains of Penicillium chrysogenum. Curr. Genet. 16, 453-459; Newbert, R.W., Barton, B., Greaves, P., Harper, J., Turner, G., 1997. Analysis of a commercially improved Penicillium chrysogenum strain series, involvement of recombinogenic regions in amplification and deletion of the penicillin gene cluster. J. Ind. Microbiol. 19, 18-27]. Bioinformatic analysis of the central 56.9kb, present as six direct repeats in the strains analyzed in this study, predicted 15 Open Reading Frames (ORFs). Besides the three penicillin biosynthetic genes (pcbAB, pcbC and penDE) only one ORF has an orthologue of known function in the database: the Saccharomyces cerevisiae gene ERG25. Surprisingly, many genes known to encode direct or indirect steps beta-lactam biosynthesis like phenyl acetic acid CoA ligase and transporters are not present. Detailed analyses reveal a detectable transcript for most of the predicted ORFs under the conditions tested. We have studied the role of these in relation to penicillin production and amplification of the biosynthetic gene cluster. In contrast to what was expected, the genes encoding the three penicillin biosynthetic enzymes alone are sufficient to restore full beta-lactam synthesis in a mutant lacking the complete region. Therefore, the role of the other 12 ORFs in this region seems irrelevant for penicillin biosynthesis.

  9. Genotypic identification of Penicillium expansum and the role of processing on patulin presence in juice.

    PubMed

    Elhariry, Hesham; Bahobial, Abdul Aziz; Gherbawy, Youssuf

    2011-04-01

    This work aimed at isolation and identification of patulin producing fungi and to follow the presence of patulin during apple juice processing. Among 34 Penicillium isolates, eight isolates (five from healthy appeared apples and 12 from rot spotted apples) were considered as patulin producers using thin-layer chromatography. These isolates were classically identified as a Penicillium expansum. PCR utilizing primers based on the polygalacturonase gene of P. expansum was applied for detecting this mold. The PCR amplified a 404-bp DNA product from all tested P. expansum isolates, but not in other common food spoilage Penicillium species. RAPD technique using P1 or M13 primers was applied to determine the similarity of the P. expansum isolates. RAPD results revealed that the tested strains showed high percentage of similarity and no correlation was observed between cluster analysis and the sources of isolation. Patulin could not be detected in healthy appeared apples and their extracted juice during different stages of juice process. In apple juice made from the healthy parts of apples decayed by P. expansum contained patulin which may present health hazard. The obtained results assured that patulin is known to be stable in apple juice even after pasteurization. In conclusion, the removal of the rotten part from the fruit is not sufficient to eliminate the mycotoxin patulin from apple juice. Although, the enzyme treatment (pectinase and amylase) and pasteurization (95 °C for 7 min) significantly (p < 0.05) reduced patulin level, its level is still higher than the level of <50 μg/kg considered by Codex alimentarius when the apple juice processed from the healthy parts of rot spotted fruits.

  10. Genetic basis for mycophenolic acid production and strain-dependent production variability in Penicillium roqueforti.

    PubMed

    Gillot, Guillaume; Jany, Jean-Luc; Dominguez-Santos, Rebeca; Poirier, Elisabeth; Debaets, Stella; Hidalgo, Pedro I; Ullán, Ricardo V; Coton, Emmanuel; Coton, Monika

    2017-04-01

    Mycophenolic acid (MPA) is a secondary metabolite produced by various Penicillium species including Penicillium roqueforti. The MPA biosynthetic pathway was recently described in Penicillium brevicompactum. In this study, an in silico analysis of the P. roqueforti FM164 genome sequence localized a 23.5-kb putative MPA gene cluster. The cluster contains seven genes putatively coding seven proteins (MpaA, MpaB, MpaC, MpaDE, MpaF, MpaG, MpaH) and is highly similar (i.e. gene synteny, sequence homology) to the P. brevicompactum cluster. To confirm the involvement of this gene cluster in MPA biosynthesis, gene silencing using RNA interference targeting mpaC, encoding a putative polyketide synthase, was performed in a high MPA-producing P. roqueforti strain (F43-1). In the obtained transformants, decreased MPA production (measured by LC-Q-TOF/MS) was correlated to reduced mpaC gene expression by Q-RT-PCR. In parallel, mycotoxin quantification on multiple P. roqueforti strains suggested strain-dependent MPA-production. Thus, the entire MPA cluster was sequenced for P. roqueforti strains with contrasted MPA production and a 174bp deletion in mpaC was observed in low MPA-producers. PCRs directed towards the deleted region among 55 strains showed an excellent correlation with MPA quantification. Our results indicated the clear involvement of mpaC gene as well as surrounding cluster in P. roqueforti MPA biosynthesis.

  11. First Report on Isolation of Penicillium adametzioides and Purpureocillium lilacinum from Decayed Fruit of Cheongsoo Grapes in Korea.

    PubMed

    Deng, Jian Xin; Paul, Narayan Chandra; Sang, Hyun Kyu; Lee, Ji Hye; Hwang, Yong Soo; Yu, Seung Hun

    2012-03-01

    Two species, Penicillium adametzioides and Purpureocillium lilacinum, were isolated from decayed grapes (cv. Cheongsoo) in Korea. Each species was initially identified by phylogenetic analysis of a combined dataset of two genes. Internal transcribed spacer (ITS) and β-tubulin (BT2) genes were used for identification of Penicillium adametzioides, and ITS and partial translation elongation factor 1-α (TEF) genes were used for identification of Purpureocillium lilacinum. Morphologically, they were found to be identical to previous descriptions. The two species presented here have not been previously reported in Korea.

  12. Sexual recombination as a tool for engineering industrial Penicillium chrysogenum strains.

    PubMed

    Dahlmann, Tim A; Böhm, Julia; Becker, Kordula; Kück, Ulrich

    2015-11-01

    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.

  13. Polyketides with Immunosuppressive Activities from Mangrove Endophytic Fungus Penicillium sp. ZJ-SY2

    PubMed Central

    Liu, Hongju; Chen, Senhua; Liu, Weiyang; Liu, Yayue; Huang, Xishan; She, Zhigang

    2016-01-01

    Nine polyketides, including two new benzophenone derivatives, peniphenone (1) and methyl peniphenone (2), along with seven known xanthones (3–9) were obtained from mangrove endophytic fungus Penicillium sp. ZJ-SY2 isolated from the leaves of Sonneratia apetala. Their structures were elucidated on the basis of MS, 1D, and 2D NMR data. Compounds 1, 3, 5, and 7 showed potent immunosuppressive activity with IC50 values ranging from 5.9 to 9.3 μg/mL. PMID:27897975

  14. Environmental and Nutritional Factors Affecting the Production of Rubratoxin B by Penicillium rubrum Stoll 1

    PubMed Central

    Hayes, A. Wallace; Wyatt, Elwanda P.; King, Patricia A.

    1970-01-01

    Rubratoxin B can be produced in a semisynthetic medium by Penicillium rubrum under varying environmental and nutritional conditions. Maximum production (552.0 mg/500 ml) was obtained with P. rubrum NRRL A-11785 grown in stationary cultures of Mosseray's simplified Raulin solution supplemented with 2.5% malt extract broth at ambient temperature. Zinc is required at levels of at least 0.4 mg per liter. In the absence of iron sulfate, there was a 50-fold reduction in rubratoxin B production but not in growth. No toxin was produced by this isolate in 5- or 7-liter fermentors. PMID:5485727

  15. Improved Mannanase Production from Penicillium occitanis by Fed-Batch Fermentation Using Acacia Seeds

    PubMed Central

    Blibech, Monia; Ellouz Ghorbel, Raoudha; Chaari, Fatma; Dammak, Ilyes; Bhiri, Fatma; Neifar, Mohamed; Ellouz Chaabouni, Semia

    2011-01-01

    By applying a fed-batch strategy, production of Penicillium occitanis mannanases could be almost doubled as compared to a batch cultivation on acacia seeds (76 versus 41 U/mL). Also, a 10-fold increase of enzyme activities was observed from shake flask fermentation to the fed-batch fermentation. These production levels were 3-fold higher than those obtained on coconut meal. The high mannanase production using acacia seeds powder as inducer substrate showed the suitability of this culture process for industrial-scale development. PMID:23724314

  16. Pretrichodermamides D–F from a Marine Algicolous Fungus Penicillium sp. KMM 4672

    PubMed Central

    Yurchenko, Anton N.; Smetanina, Olga F.; Ivanets, Elena V.; Kalinovsky, Anatoly I.; Khudyakova, Yuliya V.; Kirichuk, Natalya N.; Popov, Roman S.; Bokemeyer, Carsten; von Amsberg, Gunhild; Chingizova, Ekaterina A.; Afiyatullov, Shamil Sh.; Dyshlovoy, Sergey A.

    2016-01-01

    Three new epidithiodiketopiperazines pretrichodermamides D–F (1–3), together with the known N-methylpretrichodermamide B (4) and pretrichodermamide С (5), were isolated from the lipophilic extract of the marine algae-derived fungus Penicillium sp. KMM 4672. The structures of compounds 1–5 were determined based on spectroscopic methods. The absolute configuration of pretrichodermamide D (1) was established by a combination of modified Mosher′s method, NOESY data, and biogenetic considerations. N-Methylpretrichodermamide B (5) showed strong cytotoxicity against 22Rv1 human prostate cancer cells resistant to androgen receptor targeted therapies. PMID:27355960

  17. Mass Spectrometric Characteristics of Prenylated Indole Derivatives from Marine-Derived Penicillium sp. NH-SL.

    PubMed

    Ding, Hui; Ding, Wanjing; Ma, Zhongjun

    2017-03-22

    Two prenylated indole alkaloids were isolated from the ethyl acetate extracts of a marine-derived fungus Penicillium sp. NH-SL and one of them exhibited potent cytotoxic activity against mouse hepa 1c1c7 cells. In order to detect other bioactive analogs, we used liquid chromatogram tandem mass spectrometry (LC-MS/MS) to analyze the mass spectrometric characteristics of the isolated compounds as well as the crude extracts. As a result, three other analogs were detected, and their structures were deduced according to the similar fragmentation patterns. This is the first systematic report on the mass spectrometric characteristics of prenylated indole derivatives.

  18. Penicillium excelsum sp. nov from the Brazil Nut Tree Ecosystem in the Amazon Basin’

    PubMed Central

    Taniwaki, Marta Hiromi; Pitt, John I.; Iamanaka, Beatriz T.; Massi, Fernanda P.; Fungaro, Maria Helena P.; Frisvad, Jens C.

    2015-01-01

    A new Penicillium species, P. excelsum, is described here using morphological characters, extrolite and partial sequence data from the ITS, β-tubulin and calmodulin genes. It was isolated repeatedly using samples of nut shells and flowers from the brazil nut tree, Bertolletia excelsa, as well as bees and ants from the tree ecosystem in the Amazon rainforest. The species produces andrastin A, curvulic acid, penicillic acid and xanthoepocin, and has unique partial β-tubulin and calmodulin gene sequences. The holotype of P. excelsum is CCT 7772, while ITAL 7572 and IBT 31516 are cultures derived from the holotype. PMID:26717519

  19. Penicillium excelsum sp. nov from the Brazil Nut Tree Ecosystem in the Amazon Basin'.

    PubMed

    Taniwaki, Marta Hiromi; Pitt, John I; Iamanaka, Beatriz T; Massi, Fernanda P; Fungaro, Maria Helena P; Frisvad, Jens C

    2015-01-01

    A new Penicillium species, P. excelsum, is described here using morphological characters, extrolite and partial sequence data from the ITS, β-tubulin and calmodulin genes. It was isolated repeatedly using samples of nut shells and flowers from the brazil nut tree, Bertolletia excelsa, as well as bees and ants from the tree ecosystem in the Amazon rainforest. The species produces andrastin A, curvulic acid, penicillic acid and xanthoepocin, and has unique partial β-tubulin and calmodulin gene sequences. The holotype of P. excelsum is CCT 7772, while ITAL 7572 and IBT 31516 are cultures derived from the holotype.

  20. Mass Spectrometric Characteristics of Prenylated Indole Derivatives from Marine-Derived Penicillium sp. NH-SL

    PubMed Central

    Ding, Hui; Ding, Wanjing; Ma, Zhongjun

    2017-01-01

    Two prenylated indole alkaloids were isolated from the ethyl acetate extracts of a marine-derived fungus Penicillium sp. NH-SL and one of them exhibited potent cytotoxic activity against mouse hepa 1c1c7 cells. In order to detect other bioactive analogs, we used liquid chromatogram tandem mass spectrometry (LC-MS/MS) to analyze the mass spectrometric characteristics of the isolated compounds as well as the crude extracts. As a result, three other analogs were detected, and their structures were deduced according to the similar fragmentation patterns. This is the first systematic report on the mass spectrometric characteristics of prenylated indole derivatives. PMID:28327529

  1. X-ray diffraction study of Penicillium Vitale catalase in the complex with aminotriazole

    NASA Astrophysics Data System (ADS)

    Borovik, A. A.; Grebenko, A. I.; Melik-Adamyan, V. R.

    2011-07-01

    The three-dimensional structure of the enzyme catalase from Penicillium vitale in a complex with the inhibitor aminotriazole was solved and refined by protein X-ray crystallography methods. An analysis of the three-dimensional structure of the complex showed that the inhibition of the enzyme occurs as a result of the covalent binding of aminotriazole to the amino-acid residue His64 in the active site of the enzyme. An investigation of the three-dimensional structure of the complex resulted in the amino-acid residues being more precisely identified. The binding sites of saccharide residues and calcium ions in the protein molecule were found.

  2. X-ray diffraction study of Penicillium Vitale catalase in the complex with aminotriazole

    SciTech Connect

    Borovik, A. A.; Grebenko, A. I.; Melik-Adamyan, V. R.

    2011-07-15

    The three-dimensional structure of the enzyme catalase from Penicillium vitale in a complex with the inhibitor aminotriazole was solved and refined by protein X-ray crystallography methods. An analysis of the three-dimensional structure of the complex showed that the inhibition of the enzyme occurs as a result of the covalent binding of aminotriazole to the amino-acid residue His64 in the active site of the enzyme. An investigation of the three-dimensional structure of the complex resulted in the amino-acid residues being more precisely identified. The binding sites of saccharide residues and calcium ions in the protein molecule were found.

  3. Isolation and partial characterization of a mutant of Penicillium funiculosum for the saccharification of straw

    SciTech Connect

    Hoffman, R.M.; Wood, T.M.

    1985-01-01

    Clearing of agar plates containing ball-milled, delignified straw has been used for screening mutants of Penicillium funiculosum IMI 87160 III. The effects of glycerol and a number of sugars on the clearing were investigated for selecting derepressed mutants. The ..beta..-glucosidase synthesis by one such mutant, C22c, in shake flasks containing straw was not repressed by 5% glycerol, whereas activities on filter paper, CM-cellulose, and p-nitrophenyl-..beta..-xylosidase were only partially derepressed; xylanase was extensively derepressed. The evidence for separate control of the enzymes involved in the solubilization of straw is discussed. 23 references.

  4. Identification of intermediates in the biosynthesis of PR toxin by Penicillium roqueforti.

    PubMed

    Riclea, Ramona; Dickschat, Jeroen S

    2015-10-05

    The sesquiterpenoid 7-epi-neopetasone was synthesized via the Wieland-Miescher ketone. The compound was identical to a previously tentatively identified headspace constituent of Penicillium roqueforti. Feeding experiments with (13) C-labeled mevalonolactone isotopomers demonstrated that oxidation at C12 and an isomerization of the C11C12 to a C7C11 double bond must occur independently and not via a C7-C11-C12 allyl radical in one step. Feeding with (11,12,13-(13) C3 )-7-epi-neopetasone resulted in labelling of the PR toxin, thus establishing this compound as a newly identified pathway intermediate.

  5. Biological control of Penicillium digitatum by Trichoderma viride on postharvest citrus fruits.

    PubMed

    Díaz Borrás, A; Vila Aguilar, R

    1990-10-01

    In previous studies it was shown that Trichoderma viride, isolated from Spanish citrus packing houses, showed antagonistic activity against Penicillium digitatum in in vitro laboratory tests. In the present in vivo studies Navelina oranges, protected with aqueous suspension of T. viride (2.5 x 10(6) to 2.5 x 10(9) spores per ml), showed an increase in resistance toward P. digitatum. Oranges, inoculated with P. digitatum, did not produce lesions after 5 days when T. viride was applied 48 h or 72 h before inoculation.

  6. Rapid inactivation of Penicillium digitatum spores using high-density nonequilibrium atmospheric pressure plasma

    SciTech Connect

    Iseki, Sachiko; Hori, Masaru; Ohta, Takayuki; Aomatsu, Akiyoshi; Ito, Masafumi; Kano, Hiroyuki; Higashijima, Yasuhiro

    2010-04-12

    A promising, environmentally safe method for inactivating fungal spores of Penicillium digitatum, a difficult-to-inactivate food spoilage microorganism, was developed using a high-density nonequilibrium atmospheric pressure plasma (NEAPP). The NEAPP employing Ar gas had a high electron density on the order of 10{sup 15} cm{sup -3}. The spores were successfully and rapidly inactivated using the NEAPP, with a decimal reduction time in spores (D value) of 1.7 min. The contributions of ozone and UV radiation on the inactivation of the spores were evaluated and concluded to be not dominant, which was fundamentally different from the conventional sterilizations.

  7. Lipase production by Penicillium restrictum using solid waste of industrial babassu oil production as substrate.

    PubMed

    Palma, M B; Pinto, A L; Gombert, A K; Seitz, K H; Kivatinitz, S C; Castilho, L R; Freire, D M

    2000-01-01

    Lipase, protease, and amylase production by Penicillium restrictum in solid-state fermentation was investigated. The basal medium was an industrial waste of babassu oil (Orbignya oleifera) production. It was enriched with peptone, olive oil, and Tween-80. The supplementation positively influenced both enzyme production and fungal growth. Media enriched with Tween-80 provided the highest protease activity (8.6 U/g), whereas those enriched with peptone and olive oil led to the highest lipase (27.8 U/g) and amylase (31.8 U/g) activities, respectively.

  8. Bridged Epipolythiodiketopiperazines from Penicillium raciborskii, an Endophytic Fungus of Rhododendron tomentosum Harmaja.

    PubMed

    Kajula, Marena; Ward, Joshua M; Turpeinen, Ari; Tejesvi, Mysore V; Hokkanen, Juho; Tolonen, Ari; Häkkänen, Heikki; Picart, Pere; Ihalainen, Janne; Sahl, Hans-Georg; Pirttilä, Anna Maria; Mattila, Sampo

    2016-04-22

    Three new epithiodiketopiperazine natural products [outovirin A (1), outovirin B (2), and outovirin C (3)] resembling the antifungal natural product gliovirin have been identified in extracts of Penicillium raciborskii, an endophytic fungus isolated from Rhododendron tomentosum. The compounds are unusual for their class in that they possess sulfide bridges between α- and β-carbons rather than the typical α-α bridging. To our knowledge, outovirin A represents the first reported naturally produced epimonothiodiketopiperazine, and antifungal outovirin C is the first reported trisulfide gliovirin-like compound. This report describes the identification and structural elucidation of the compounds by LC-MS/MS and NMR.

  9. Penicillium arizonense, a new, genome sequenced fungal species, reveals a high chemical diversity in secreted metabolites

    PubMed Central

    Grijseels, Sietske; Nielsen, Jens Christian; Randelovic, Milica; Nielsen, Jens; Nielsen, Kristian Fog; Workman, Mhairi; Frisvad, Jens Christian

    2016-01-01

    A new soil-borne species belonging to the Penicillium section Canescentia is described, Penicillium arizonense sp. nov. (type strain CBS 141311T = IBT 12289T). The genome was sequenced and assembled into 33.7 Mb containing 12,502 predicted genes. A phylogenetic assessment based on marker genes confirmed the grouping of P. arizonense within section Canescentia. Compared to related species, P. arizonense proved to encode a high number of proteins involved in carbohydrate metabolism, in particular hemicellulases. Mining the genome for genes involved in secondary metabolite biosynthesis resulted in the identification of 62 putative biosynthetic gene clusters. Extracts of P. arizonense were analysed for secondary metabolites and austalides, pyripyropenes, tryptoquivalines, fumagillin, pseurotin A, curvulinic acid and xanthoepocin were detected. A comparative analysis against known pathways enabled the proposal of biosynthetic gene clusters in P. arizonense responsible for the synthesis of all detected compounds except curvulinic acid. The capacity to produce biomass degrading enzymes and the identification of a high chemical diversity in secreted bioactive secondary metabolites, offers a broad range of potential industrial applications for the new species P. arizonense. The description and availability of the genome sequence of P. arizonense, further provides the basis for biotechnological exploitation of this species. PMID:27739446

  10. Metabolomic and Transcriptomic Comparison of Solid-State and Submerged Fermentation of Penicillium expansum KACC 40815.

    PubMed

    Kim, Hyang Yeon; Heo, Do Yeon; Park, Hye Min; Singh, Digar; Lee, Choong Hwan

    2016-01-01

    Penicillium spp. are known to harbor a wide array of secondary metabolites with cryptic bioactivities. However, the metabolomics of these species is not well-understood in terms of different fermentation models and conditions. The present study involved metabolomics profiling and transcriptomic analysis of Penicillium expansum 40815 under solid-state fermentation (SSF) and submerged fermentation (SmF). Metabolite profiling was carried out using ultra-performance liquid chromatography quadruple time-of-flight mass spectrometry with multivariate analysis, followed by transcriptomic analyses of differentially expressed genes. In principal component analysis, the metabolite profiling data was studied under different experimental sets, including SSF and SmF. The significantly different metabolites such as polyketide metabolites (agonodepside B, rotiorin, verrucosidin, and ochrephilone) and corresponding gene transcripts (polyketide synthase, aromatic prenyltransferase, and terpenoid synthase) were primarily detected under SmF conditions. In contrast, the meroterpenoid compounds (andrastin A and C) and their genes transcripts were exclusively detected under SSF conditions. We demonstrated that the metabolite production and its corresponding gene expression levels in P. expansum 40815 were significantly influenced by the varying growth parameters and the immediate environment. This study further provides a foundation to produce specific metabolites by regulating fermentation conditions.

  11. Penicillium sp. as an organism that degrades endosulfan and reduces its genotoxic effects.

    PubMed

    Romero-Aguilar, Mariana; Tovar-Sánchez, Efrain; Sánchez-Salinas, Enrique; Mussali-Galante, Patricia; Sánchez-Meza, Juan Carlos; Castrejón-Godínez, María Luisa; Dantán-González, Edgar; Trujillo-Vera, Miguel Ángel; Ortiz-Hernández, Ma Laura

    2014-01-01

    Endosulfan is an organochloride and persistent pesticide that has caused concern because of its impact in the environment and its toxicity to and bioaccumulation in living organisms. In this study, we isolated an endosulfan-degrading fungus from the activated sludge from an industrial wastewater treatment plant. Through repetitive enrichment and successive subculture in media containing endosulfan as the sole carbon source, a fungus designated CHE 23 was isolated. Based on a phylogenetic analysis, strain CHE 23 was assigned to the genus Penicillium sp. In a mineral salt medium with 50 mg/l endosulfan as the sole source carbon, CHE 23 removed the added endosulfan in a period of six days. To verify the decrease in endosulfan toxicity due to the activity of the fungus, we performed genotoxicity tests trough the single cell gel electrophoresis assay or comet assay, with Eisenia fetida as the bioindicator species. This organism was exposed to the supernatants of the culture of the fungus and endosulfan. Our results indicated that the genotoxicity of endosulfan was completely reduced due the activity of this fungus. These results suggest that the Penicillium sp. CHE 23 strain can be used to degrade endosulfan residues and/or for water and soil bioremediation processes without causing toxicity problems, which are probably due to the generation of no-toxic metabolites during biodegradation.

  12. Purification and characterization of an extracellular protease from Penicillium chrysogenum Pg222 active against meat proteins.

    PubMed

    Benito, María J; Rodríguez, Mar; Núñez, Félix; Asensio, Miguel A; Bermúdez, María E; Córdoba, Juan J

    2002-07-01

    An extracellular protease from Penicillium chrysogenum (Pg222) isolated from dry-cured ham has been purified. The purification procedure involved several steps: ammonium sulfate precipitation, ion-exchange chromatography, filtration, and separation by high-performance liquid chromatography. Based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis and gel filtration, the purified fraction showed a molecular mass of about 35 kDa. The hydrolytic properties of the purified enzyme (EPg222) on extracted pork myofibrillar proteins under several conditions were evaluated by SDS-PAGE. EPg222 showed activity in the range of 10 to 60 degrees C in temperature, 0 to 3 M NaCl, and pH 5 to 7, with maximum activity at pH 6, 45 degrees C, and 0.25 M NaCl. Under these conditions the enzyme was most active against tropomyosin, actin, and myosin. EPg222 showed collagenolytic activity but did not hydrolyze myoglobin. EPg222 showed higher activity than other proteolytic enzymes like papain, trypsin, and Aspergillus oryzae protease. The N-terminal amino acid sequence was determined and was found to be Glu-Asn-Pro-Leu-Gln-Pro-Asn-Ala-Pro-Ser-Trp. This partial amino acid sequence revealed a 55% homology with serine proteases from Penicillium citrinum. The activity of this novel protease may be of interest in ripening and generating the flavor of dry-cured meat products.

  13. Hydrolytic activity of Penicillium chrysogenum Pg222 on pork myofibrillar proteins.

    PubMed

    Benito, María J; Córdoba, Juan J; Alonso, Mercedes; Asensio, Miguel A; Núñez, Félix

    2003-12-31

    Moulds grow on many different dry-cured meat products and are able to hydrolyse muscle proteins. However, their contribution to proteolysis in these products is not well known. Only recently, the ability of just a few strains of Penicillium spp. to increase proteolysis in dry-cured meat products has been shown. For these strains to be used as starter cultures, their hydrolytic activity under standard conditions should be characterised. With this purpose, the effect of Penicillium chrysogenum Pg222 on pork myofibrillar proteins has been assayed in a culture medium containing 5% (w/v) NaCl. SDS-PAGE revealed that Pg222 was responsible for extensive hydrolysis of the main myofibrillar proteins except alpha-actinin. The proteolysis led to increases in free amino acids, reaching peak values at 84 h. Ala, Tyr and Lys were present in the greatest amount. These results suggest that P. chrysogenum Pg222 would contribute to development of desired texture and flavours in dry-cured meat products.

  14. Purification and Characterization of an Extracellular Protease from Penicillium chrysogenum Pg222 Active against Meat Proteins

    PubMed Central

    Benito, María J.; Rodríguez, Mar; Núñez, Félix; Asensio, Miguel A.; Bermúdez, María E.; Córdoba, Juan J.

    2002-01-01

    An extracellular protease from Penicillium chrysogenum (Pg222) isolated from dry-cured ham has been purified. The purification procedure involved several steps: ammonium sulfate precipitation, ion-exchange chromatography, filtration, and separation by high-performance liquid chromatography. Based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis and gel filtration, the purified fraction showed a molecular mass of about 35 kDa. The hydrolytic properties of the purified enzyme (EPg222) on extracted pork myofibrillar proteins under several conditions were evaluated by SDS-PAGE. EPg222 showed activity in the range of 10 to 60°C in temperature, 0 to 3 M NaCl, and pH 5 to 7, with maximum activity at pH 6, 45°C, and 0.25 M NaCl. Under these conditions the enzyme was most active against tropomyosin, actin, and myosin. EPg222 showed collagenolytic activity but did not hydrolyze myoglobin. EPg222 showed higher activity than other proteolytic enzymes like papain, trypsin, and Aspergillus oryzae protease. The N-terminal amino acid sequence was determined and was found to be Glu-Asn-Pro-Leu-Gln-Pro-Asn-Ala-Pro-Ser-Trp. This partial amino acid sequence revealed a 55% homology with serine proteases from Penicillium citrinum. The activity of this novel protease may be of interest in ripening and generating the flavor of dry-cured meat products. PMID:12089038

  15. Cloning and heterologous expression of a thermostable pectate lyase from Penicillium occitanis in Escherichia coli.

    PubMed

    Damak, Naourez; Abdeljalil, Salma; Koubaa, Aida; Trigui, Sameh; Ayadi, Malika; Trigui-Lahiani, Hèla; Kallel, Emna; Turki, Nadia; Djemal, Lamia; Belghith, Hafeth; Taieb, Noomen Hadj; Gargouri, Ali

    2013-11-01

    The entire pectate lyase cDNA (Pel1) of Penicillium occitanis was cloned from a cDNA bank and sequenced. The ORF exhibited a great homology to Penicillium marneffei and conservation of all features of fungal pectate lyases such as the barrel structure with "eight right-handed parallel β-helix" architecture. The structure modeling also showed the interesting resemblance with thermostable pectate lyases since several specific residues were also shared by Pel1 and these thermostable enzymes. Having shown that the enzyme retains its activity after endoH-mediated deglycosylation, we investigated its expression in Escherichia coli BL21 using the pET28-a vector. This expression was shown to be optimum when cells were induced at room temperature in 2YT medium rather than at 37 °C and LB medium. In such conditions, the recombinant protein was apparently produced more in soluble form than as inclusion bodies. The effect of NaCl concentration was investigated during the binding and elution steps of recombinant His-tagged enzyme on MagneHis Ni-particles. The purified enzyme was shown to retain its thermo-activity as well as a great tolerance to high concentration of NaCl and imidazole.

  16. Penicillium glabrum cork-colonising isolates - preliminary analysis of their genomic similarity.

    PubMed

    Basílio, Maria Carmo; Gaspar, Ricardo; Silva Pereira, Cristina; San Romão, Maria Vitória

    2006-09-01

    The cork stopper manufacturing process includes an operation, known as stabilisation, by which humid cork slabs are extensively colonised by fungi. The effects of fungal growth on cork are not completely understood although they are considered to be involved in the so-called "cork taint" of wine. It is essential to (a) identify environmental constraints which define the appearance of the colonising fungal species and (b) trace their origin to the forest and/or the manufacturing space. The present article correlates two sets of data, from consecutive years and the same season, of systematic sampling of two manufacturing units, located in the North and South of Portugal. Chrysonilia sitophila dominance was confirmed, followed by a high diversity of Penicillium species. Penicillium glabrum, which was found in all samples, was the most frequently isolated species. P. glabrum intra-species variability was investigated using DNA fingerprinting techniques revealing highly discriminative polymorphic markers in the genome. Cluster analysis of P. glabrum data was discussed in relation to the geographical location of strains, and results suggest that P. glabrum arise from predominantly the manufacturing space, although cork specific fungi can contribute.

  17. Identification of a Polyketide Synthase Involved in Sorbicillin Biosynthesis by Penicillium chrysogenum

    PubMed Central

    Salo, Oleksandr; Guzmán-Chávez, Fernando; Ries, Marco I.; Lankhorst, Peter P.; Bovenberg, Roel A. L.; Vreeken, Rob J.

    2016-01-01

    ABSTRACT Secondary metabolism in Penicillium chrysogenum was intensively subjected to classical strain improvement (CSI), the resulting industrial strains producing high levels of β-lactams. During this process, the production of yellow pigments, including sorbicillinoids, was eliminated as part of a strategy to enable the rapid purification of β-lactams. Here we report the identification of the polyketide synthase (PKS) gene essential for sorbicillinoid biosynthesis in P. chrysogenum. We demonstrate that the production of polyketide precursors like sorbicillinol and dihydrosorbicillinol as well as their derivatives bisorbicillinoids requires the function of a highly reducing PKS encoded by the gene Pc21g05080 (pks13). This gene belongs to the cluster that was mutated and transcriptionally silenced during the strain improvement program. Using an improved β-lactam-producing strain, repair of the mutation in pks13 led to the restoration of sorbicillinoid production. This now enables genetic studies on the mechanism of sorbicillinoid biosynthesis in P. chrysogenum and opens new perspectives for pathway engineering. IMPORTANCE Sorbicillinoids are secondary metabolites with antiviral, anti-inflammatory, and antimicrobial activities produced by filamentous fungi. This study identified the gene cluster responsible for sorbicillinoid formation in Penicillium chrysogenum, which now allows engineering of this diverse group of compounds. PMID:27107123

  18. Metabolomic and Transcriptomic Comparison of Solid-State and Submerged Fermentation of Penicillium expansum KACC 40815

    PubMed Central

    Park, Hye Min; Singh, Digar; Lee, Choong Hwan

    2016-01-01

    Penicillium spp. are known to harbor a wide array of secondary metabolites with cryptic bioactivities. However, the metabolomics of these species is not well-understood in terms of different fermentation models and conditions. The present study involved metabolomics profiling and transcriptomic analysis of Penicillium expansum 40815 under solid-state fermentation (SSF) and submerged fermentation (SmF). Metabolite profiling was carried out using ultra-performance liquid chromatography quadruple time-of-flight mass spectrometry with multivariate analysis, followed by transcriptomic analyses of differentially expressed genes. In principal component analysis, the metabolite profiling data was studied under different experimental sets, including SSF and SmF. The significantly different metabolites such as polyketide metabolites (agonodepside B, rotiorin, verrucosidin, and ochrephilone) and corresponding gene transcripts (polyketide synthase, aromatic prenyltransferase, and terpenoid synthase) were primarily detected under SmF conditions. In contrast, the meroterpenoid compounds (andrastin A and C) and their genes transcripts were exclusively detected under SSF conditions. We demonstrated that the metabolite production and its corresponding gene expression levels in P. expansum 40815 were significantly influenced by the varying growth parameters and the immediate environment. This study further provides a foundation to produce specific metabolites by regulating fermentation conditions. PMID:26863302

  19. Purification, sequencing and evaluation of a divergent phytase from Penicillium oxalicum KCTC6440.

    PubMed

    Kim, Bong-Hyun; Lee, Ji Yeon; Lee, Peter C W

    2015-01-01

    A fungal strain producing high levels of phytase was purified to homogeneity from Penicillium oxalicum KCTC6440 (PhyA). The molecular mass of the purified PhyA was 65 kDa and optimal activity occurred at 55°C. The enzyme was stable in a pH range of 4.5-6.5, with an optimum performance at pH 5.5. The Km value for the substrate sodium phytate was 0.48 mM with a Vmax of 672 U/mg. The enzyme was inhibited by Ca(2+), Cu(2+), and Zn(2+), and slightly enhanced by EDTA. The PhyA efficiently released phosphate from feedstuffs such as soybean, rich bran and corn meal. The PhyA gene was cloned in two steps of degenerate PCR and inverse PCR and found to comprise 1501 bp and encode 461 amino acid residues. The enzyme was found to have only 13 amino acids differing to the known PhyA from other Penicillium sp., but has distinct enzyme characteristics. Computational analysis showed that PhyA possessed more positively charged residues in the active sites compared to other PhyA molecules, which may explain the broader pH spectrum.

  20. Proteomics Insights into the Biomass Hydrolysis Potentials of a Hypercellulolytic Fungus Penicillium funiculosum.

    PubMed

    Ogunmolu, Funso Emmanuel; Kaur, Inderjeet; Gupta, Mayank; Bashir, Zeenat; Pasari, Nandita; Yazdani, Syed Shams

    2015-10-02

    The quest for cheaper and better enzymes needed for the efficient hydrolysis of lignocellulosic biomass has placed filamentous fungi in the limelight for bioprospecting research. In our search for efficient biomass degraders, we identified a strain of Penicillium funiculosum whose secretome demonstrates high saccharification capabilities. Our probe into the secretome of the fungus through qualitative and label-free quantitative mass spectrometry based proteomics studies revealed a high abundance of inducible CAZymes and several nonhydrolytic accessory proteins. The preferential association of these proteins and the attending differential biomass hydrolysis gives an insight into their interactions and clues about possible roles of novel hydrolytic and nonhydrolytic proteins in the synergistic deconstruction of lignocellulosic biomass. Our study thus provides the first comprehensive insight into the repertoire of proteins present in a high-performing secretome of a hypercellulolytic Penicillium funiculosum, their relative abundance in the secretome, and the interaction dynamics of the various protein groups in the secretome. The gleanings from the stoichiometry of these interactions hold a prospect as templates in the design of cost-effective synthetic cocktails for the optimal hydrolysis of biomass.

  1. Rapid extra-/intracellular biosynthesis of gold nanoparticles by the fungus Penicillium sp.

    NASA Astrophysics Data System (ADS)

    Du, Liangwei; Xian, Liang; Feng, Jia-Xun

    2011-03-01

    In this work, the fungus Penicillium was used for rapid extra-/intracellular biosynthesis of gold nanoparticles. AuCl4 - ions reacted with the cell filtrate of Penicillium sp. resulting in extracellular biosynthesis of gold nanoparticles within 1 min. Intracellular biosynthesis of gold nanoparticles was obtained by incubating AuCl4 - solution with fungal biomass for 8 h. The gold nanoparticles were characterized by means of visual observation, UV-Vis absorption spectroscopy, X-ray diffraction (XRD), transmission electron microscopy (TEM), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDX). The extracellular nanoparticles exhibited maximum absorbance at 545 nm in UV-Vis spectroscopy. The XRD spectrum showed Bragg reflections corresponding to the gold nanocrystals. TEM exhibited the formed spherical gold nanoparticles in the size range from 30 to 50 nm with an average size of 45 nm. SEM and TEM revealed that the intracellular gold nanoparticles were well dispersed on the cell wall and within the cell, and they are mostly spherical in shape with an average diameter of 50 nm. The presence of gold was confirmed by EDX analysis.

  2. Penicillium salamii strain ITEM 15302: A new promising fungal starter for salami production.

    PubMed

    Magistà, D; Ferrara, M; Del Nobile, M A; Gammariello, D; Conte, A; Perrone, G

    2016-08-16

    Traditional sausages are often considered of superior quality to sausages inoculated with commercial starter cultures and this is partially due to the action of the typical house microflora. Penicillium nalgiovense is the species commonly used as starter culture for dry-cured meat production. Recently a new species, Penicillium salamii, was described as typical colonizer during salami seasoning. In order to understand its contribution to the seasoning process, two different experiments on curing of fresh pork sausages were conducted using P. salamii ITEM 15302 in comparison with P. nalgiovense ITEM 15292 at small and industrial scale, and the dry-cured sausages were subjected to sensory analyses. Additionally, proteolytic and lipolytic in vitro assays were performed on both strains. P. salamii ITEM 15302 proved to be a fast growing mould on dry-cured sausage casings, well adapted to the seasoning process, with high lipolytic and proteolytic enzymatic activity that confers typical sensory characteristics to meat products. Therefore, P. salamii ITEM 15302 was shown to be a good candidate as new starter for meat industry.

  3. Aerobiological analysis in a salami factory: a possible case of extrinsic allergic alveolitis by Penicillium camembertii.

    PubMed

    Marchisio, V F; Sulotto, F; Botta, G C; Chiesa, A; Airaudi, D; Anastasi, A

    1999-08-01

    A 39-year-old man was hospitalized with a history of fatigue, dyspnoea and low grade fever which seemed to be related to his working environment. The patient was employed in a salami factory, working near the area where the salami are seasoned with fungal inocula. Chest X-ray showed diffuse initial changes of reticulonodular pattern that disappeared after a brief course of steroids therapy. Precipitating antibodies to Penicillium notatum and Aspergillus fumigatus were found both in plasma and bronchoalveolar lavage fluid. This, together with the finding of a lymphocytic alveolitis with CD4+ depletion and CD8+ increase, suggested the possibility of extrinsic allergic alveolitis of fungal aetiology. Qualitative and quantitative monitoring with an impinger of both the working and outside environment for aerial fungal concentration demonstrated a very high level of contamination (up to 1.14x10(9) fungal propagules m-3 of air) and an inside/outside ratio from 21 to about 2000. Penicillium camembertii was the most common species found in all the indoor sites (60-100% of the fungal load). The patient's BALF and serum both displayed precipitating antibodies to P. camembertii from the powder used for the inoculum and the air samples. These results together with the patient's working history gave some evidence of relationship between the indoor P. camembertii concentration and the patient's symptoms.

  4. Comparative Secretome Analysis of Aspergillus niger, Trichoderma reesei, and Penicillium oxalicum During Solid-State Fermentation.

    PubMed

    Gong, Weili; Zhang, Huaiqiang; Liu, Shijia; Zhang, Lili; Gao, Peiji; Chen, Guanjun; Wang, Lushan

    2015-11-01

    Filamentous fungi such as Aspergillus spp., Trichoderma spp., and Penicillium spp. are frequently used to produce high concentrations of lignocellulosic enzymes. This study examined the discrepancies in the compositions and dynamic changes in the extracellular enzyme systems secreted by Aspergillus niger ATCC1015, Trichoderma reesei QM9414, and Penicillium oxalicum 114-2 cultured on corn stover and wheat bran. The results revealed different types and an abundance of monosaccharides and oligosaccharides were released during incubation, which induced the secretion of diverse glycoside hydrolases. Both the enzyme activities and isozyme numbers of the three fungal strains increased with time. A total of 279, 161, and 183 secretory proteins were detected in A. niger, T. reesei, and P. oxalicum secretomes, respectively. In the A. niger secretomes, more enzymes involved in the degradation of (galacto)mannan, xyloglucan, and the backbone of pectin distributed mostly in dicots were detected. In comparison, although P. oxalicum 114-2 hardly secreted any xyloglucanases, the diversities of enzymes involved in the degradation of xylan and β-(1,3;1,4)-D-glucan commonly found in monocots were higher. The cellulase system of P. oxalicum 114-2 was more balanced. The degradation preference provided a new perspective regarding the recomposition of lignocellulosic enzymes based on substrate types.

  5. Identification and characterization of chitin synthase genes in the postharvest citrus fruit pathogen Penicillium digitatum.

    PubMed

    Gandía, Mónica; Harries, Eleonora; Marcos, Jose F

    2012-06-01

    In this study, we carried out the isolation and characterization of chitin synthase genes (CHS) of the main citrus fruit postharvest pathogen Penicillium digitatum. Using distinct sets of degenerate primers designed from conserved regions of CHS genes of yeast and filamentous fungi, PCR methods, and a DNA genomic library, five putative CHS genes (PdigCHSI, PdigCHSII, PdigCHSIII, PdigCHSV, and PdigCHSVII) were identified, isolated, sequenced, and characterized. Phylogenetic analyses, sequence identity, and domain conservation support the annotation as CHS. A very high sequence identity and strong synteny were found with corresponding regions from the genome of Penicillium chrysogenum. Gene expression of P. digitatum CHS genes during mycelium axenic growth, under oxidative and osmotic stress conditions, and during infection of citrus fruits was confirmed and quantified using quantitative RT-PCR (qRT-PCR). PdigCHSIII had the highest expression among the five genes by one order of magnitude, while PdigCHSII had the lowest. However, PdigCHSII was strongly induced coincident with conidial production, suggesting a role in conidiogenesis. The expression of PdigCHSI, PdigCHSIII, PdigCHSV, and PdigCHSVII was upregulated during infection of citrus fruit. PdigCHSV and PdigCHSVII coexpressed in most of the experiments carried out, and they are separated by a 1.77 kb intergenic region and arranged in opposite directions.

  6. [Purification and characterization of a novel alpha-galactosidase from penicillium sp. F63 CGMCC1669].

    PubMed

    Mi, Shi-jun; Bai, Ying-guo; Meng, Kun; Wang, Ya-ru; Yao, Bin; Shi, Xiu-yun; Huang, Huo-qing; Zhang, Yu-hong; Shi, Peng-jun

    2007-02-01

    An a-galactosidase-producing fungus was screened out of 26 filamentous fungi isolated from soil by us. Phylogenetic analysis based on the alignment of 18S rDNA sequences, combined with the morphological identification, indicated that the strain F63 was a member of the genus Penicillium. The a-galactosidase from Penicillium sp. F63 was purified to apparent homogeneity by ammonium sulfate precipitation, ion-exchange and gel filtration chromatography. The molecular size of the purified enzyme is approximately 82kDa estimated by SDS-PAGE. The a-galactosidase has an optimum pH of 5.0 and an optimum temperature of 45 degrees C. The enzyme is stable between pH5.0 and 6.0 below 40 degrees C. The a-galactosidase activity is slightly inhibited by Ag+ , which is dissimilar to other a-galactosidases. Kinetic studies of the a-galactosidase showed that the Km and the Vmax for pNPG are 1.4mmol/L and 1.556mmol/L. min(-1) x mg- 1, respectively. The enzyme is able to degrade natural substrates such as melibiose, raffinose and stachyose but not galactose-containing polysaccharides. The alpha-galactosidase was identified by MALDI-TOF-MS and its inner peptides were sequenced by ESI-MS/MS. The results show that the a-galactosidase is a novel one.

  7. Effect of ozone on sterilization of Penicillium digitatum using non-equilibrium atmospheric pressure plasma

    NASA Astrophysics Data System (ADS)

    Ohta, Takayuki; Iseki, Sachiko; Ito, Masafumi; Kano, Hiroyuki; Higashijima, Yasuhiro; Hori, Masaru

    2008-10-01

    Methyl bromide has been sprayed to the crops for protecting from insects and virus, but has high ozone depletion potential. Thus, the development of substitute-technology has been strongly required. We have investigated a plasma sterilization for spores of Penicillium digitatum, which causes green mold disease of the crops, using non-equilibrium atmospheric pressure plasma. The sterilization was caused by UV light, ozone, O and OH radicals. In this study, ozone density was measured and the effect to sterilization was discussed. The plasma was generated at an alternative current of 6kV and Ar gas flow rate of 3L/min. In order to investigate the sterilization mechanism of ozone, the absolute density of ozone was measured using ultraviolet absorption spectroscopy and was from 2 to 8 ppm. The sterilization by this plasma was larger than that by the ozonizer (03:600ppm). It is confirmed that the effect of ozone to the sterilization of Penicillium digitatum would be small.

  8. GenBank submission of draft whole genome sequence of the apple decay pathogen Penicillium solitum (RS1 isolate)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Penicillium species cause postharvest blue mold decay of apples and pears in the United States and in many countries worldwide. This genus is responsible for severe economic losses and produces an array of mycotoxins that contaminate processed apple products. Among the species that cause blue mold,...

  9. Speciation despite globally overlapping distributions in Penicillium chrysogenum: the population genetics of Alexander Fleming’s lucky fungus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Eighty years ago, Alexander Fleming described the antibiotic effects of a fungus that had contaminated his bacterial culture, kick starting the antimicrobial revolution. The fungus was later ascribed to a globally distributed asexual species, Penicillium chrysogenum. Recently, the species has been...

  10. Molecular characterization and a multiplex allele-specific PCR method for detection of thiabendazole resistance in Penicillium expansum from apple

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Thiabendazole (TBZ) is commonly used as a postharvest treatment for control of blue mold in apples caused by Penicillium expansum. Different point mutations in the ß-tubulin gene conferring benzimidazole resistance have been reported in plant pathogens, but molecular mechanisms of TBZ resistance in ...

  11. GenBank submission of draft whole genome sequence of the apple decay pathogen Penicillium expansum isolate (R19)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Penicillium species cause postharvest blue mold decay of apple and pear fruits in the United States and around the world. This genus is responsible for severe economic losses and produces an array of mycotoxins that contaminate processed apple products. Among the species that cause blue mold, isolat...

  12. Carbon, nitrogen and pH regulate the production and activity of a polygalacturonase isozyme produced by Penicillium expansum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The influence of carbon, nitrogen and pH on polygalacturonase activity produced by Penicillium expansum were investigated. P. expansum mycelial growth was greatest on lyophilized fruit tissue and the highest PG activity occurred in apple pectin medium. Nitrogen source influenced PG activity and was ...

  13. Identification of wild apple germplasm (Malus spp.) with resistance to the postharvest decay pathogens Penicillium expansum and Colletotrichum acutatum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Penicillium expansum and Colletotrichum acutatum cause blue mold and bitter rot of apples during storage which results in significant economic losses. Resistance to these pathogens in commercial apple cultivars has not been documented in the literature. An apple germplasm collection, from the center...

  14. Purification and biochemical characterization of polygalacturonase produced by Penicillium expansum during postharvest decay of ‘Anjou’ pear

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A polygalacturonase (PG) was extracted and purified from decayed tissue of ‘Anjou’ pear fruit inoculated with Penicillium expansum. Ammonium sulfate precipitation, gel filtration and cation exchange chromatography were used to purify the enzyme. Both chromatographic methods revealed a single peak co...

  15. Radiosensitization of Aspergillus niger and Penicillium chrysogenum using basil essential oil and ionizing radiation for food decontamination.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Minimum Inhibitory Concentration (MIC) of basil oil, was determined for two pathogenic fungi of rice, Aspergillus niger and Penicillium chrysogenum. The antifungal activity of the basil oil in combination with ionising radiation was then investigated to determine if basil oil caused radiosensit...

  16. First report of Penicillium expansum isolates with reduced sensitivity to fludioxonil from a commercial packinghouse in Pennsylvania

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Blue mold is caused by Penicillium expansum and is among the most economically significant disease of stored apples worldwide. The fungus gains ingress through cracks, natural openings, and wounds in the fruit and produces mycotoxins that contaminate processed apple products. All commercial apples a...

  17. Preliminary evaluation of apple germplasm from Kazakhstan for resistance to blue mold decay caused by Penicillium expansum after harvest

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Blue mold of apples, incited by Penicillium expansum, causes extensive loss on stored apples worldwide. Despite the severity of this problem, apple breeders do not evaluate their crosses for resistance to this disease, because there has been little resistance to blue mold in the gene pool of the ge...

  18. Isolation, purification, and characterization of a polygalacturonase produced in Penicillium solitum-decayed 'Golden Delicious' apple fruit

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Polygalacturonase (PG) was extracted and purified from decayed Golden Delicious apple fruit inoculated with Penicillium solitum. Gel filtration and cation exchange chromatography were used to purify the enzyme. Both methods revealed a single peak corresponding to PG activity and analysis of cation ...

  19. Penicillium solitum produces a polygalacturonase isozyme in decayed ‘Anjou’ pear fruit capable of macerating host tissue in vitro

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A polygalacturonase (PG) isozyme was isolated from Penicillium solitum-decayed ‘Anjou’ pear fruit and purified to homogeneity using a multistep process. Both gel filtration and cation exchange chromatography revealed a single PG activity peak and analysis of the purified protein showed a single band...

  20. Polymerase Chain Reaction (PCR)-based methods for detection and identification of mycotoxigenic Penicillium species using conserved genes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Polymerase chain reaction amplification of conserved genes and sequence analysis provides a very powerful tool for the identification of toxigenic as well as non-toxigenic Penicillium species. Sequences are obtained by amplification of the gene fragment, sequencing via capillary electrophoresis of d...

  1. Simultaneous Saccharification and fermentation of cellulose to ethanol using Penicillium funiculosum cellulose and free or immobilized Saccharomyces uvarum cells

    SciTech Connect

    Deshpa V.; Sivaraman, H.; Rao, M.

    1983-06-01

    This communication discusses the compatibility of Penicillium funiculosum cellulase with Saccharomyces uvarum cells and the results on the combined hydrolysis and fermentation of cellulose to ethanol using a system of S. uvarum cells immobilized in an open pore gelatin matrix described earlier from the same laboratory. (Refs. 10).

  2. Comparative stability and catalytic and chemical properties of the sulfate-activating enzymes from Penicillium chrysogenum (mesophile) and Penicillium duponti (thermophile).

    PubMed

    Renosto, F; Schultz, T; Re, E; Mazer, J; Chandler, C J; Barron, A; Segel, I H

    1985-11-01

    ATP sulfurylases from Penicillium chrysogenum (a mesophile) and from Penicillium duponti (a thermophile) had a native molecular weight of about 440,000 and a subunit molecular weight of about 69,000. (The P. duponti subunit appeared to be a little smaller than the P. chrysogenum subunit.) The P. duponti enzyme was about 100 times more heat stable than the P. chrysogenum enzyme; k inact (the first-order rate constant for inactivation) at 65 degrees C = 3.3 X 10(-4) s-1 for P. duponti and 3.0 X 10(-2) s-1 for P. chrysogenum. The P. duponti enzyme was also more stable to low pH and urea at 30 degrees C. Rabbit serum antibodies to each enzyme showed heterologous cross-reaction. Amino acid analyses disclosed no major compositional differences between the two enzymes. The analogous Km and Ki values of the forward and reverse reactions were also essentially identical at 30 degrees C. At 30 degrees C, the physiologically important adenosine 5'-phosphosulfate (APS) synthesis activity of the P. duponti enzyme was 4 U mg of protein-1, which is about half that of the P. chrysogenum enzyme. The molybdolysis and ATP synthesis activities of the P. duponti enzyme at 30 degrees C were similar to those of the P. chrysogenum enzyme. At 50 degrees C, the APS synthesis activity of the P. duponti enzyme was 12 to 19 U mg of protein-1, which was higher than that of the P. chrysogenum enzyme at 30 degrees C (8 +/- 1 U mg of protein-1). Treatment of the P. chrysogenum enzyme with 5,5'-dithiobis(2-nitrobenzoate) (DTNB) at 30 degrees C under nondenaturing conditions modified one free sulfhydryl group per subunit. Vmax was not significantly altered, but the catalytic activity at low magnesium-ATP or SO4(2-) (or MoO4(2-)) was markedly reduced. Chemical modification with tetranitromethane had the same results on the kinetics. The native P. duponti enzyme was relatively unreactive toward DTNB or tetranitromethane at 30 degrees C and pH 8.0 or pH 9.0, but at 50 degrees C and pH 8.0, DTNB rapidly

  3. Pulmonary fungus ball caused by Penicillium capsulatum in a patient with type 2 diabetes: a case report

    PubMed Central

    2013-01-01

    Background Following the recent transfer of all accepted species of Penicillium subgenus Biverticillium to Talaromyces (including Talaromyces marneffei, formerly Penicillium marneffei), Penicillium species are becoming increasingly rare causal agents of invasive infections. Herein, we present a report of a type 2 diabetes patient with a fungus ball in the respiratory tract caused by Penicillium capsulatum. Case presentation A 56-year-old Chinese female gardener with a 5-year history of type 2 diabetes presented at the Shanghai Changzheng Hospital with fever, a cough producing yellow-white sputum, and fatigue. The therapeutic effect of cefoxitin was poor. An HIV test was negative, but the β-D-glucan test was positive (459.3 pg/ml). Chest radiography revealed a cavitary lesion in the left upper lobe, and a CT scan showed globate cavities with a radiopaque, gravity-dependent ball. The histopathologic features of the tissue after haematoxylin-eosin staining showed septate hyphae. The fungus was isolated from the gravity-dependent ball and identified as Penicillium capsulatum based on the morphological analysis of microscopic and macroscopic features and on ribosomal internal transcribed spacer sequencing. After surgery, the patient was cured with a sequential treatment of fluconazole 400 mg per day for 90 days and caspofungin 70 mg per day for 14 days. Conclusions Although the prognosis is often satisfactory, clinicians, mycologists and epidemiologists should be aware of the possibility of infection by this uncommon fungal pathogen in diabetes patients, since it may cause severe invasive infections in immunocompromised hosts such as diabetes and AIDS patients. PMID:24152579

  4. The development of genetic and molecular markers to register and commercialize Penicillium rubens (formerly Penicillium oxalicum) strain 212 as a biocontrol agent.

    PubMed

    Villarino, Maria; De Cal, Antonieta; Melgarejo, Paloma; Larena, Inmaculada; Espeso, Eduardo A

    2016-01-01

    Penicillium oxalicum strain 212 (PO212) is an effective biocontrol agent (BCA) against a large number of economically important fungal plant pathogens. For successful registration as a BCA in Europe, PO212 must be accurately identified. In this report, we describe the use of classical genetic and molecular markers to characterize and identify PO212 in order to understand its ecological role in the environment or host. We successfully generated pyrimidine (pyr-) auxotrophic mutants. In addition we also designed specific oligonucleotides for the pyrF gene at their untranslated regions for rapid and reliable identification and classification of strains of P. oxalicum and P. rubens, formerly P. chrysogenum. Using these DNA-based technologies, we found that PO212 is a strain of P. rubens, and is not a strain of P. oxalicum. This work presents PO212 as the unique P. rubens strain to be described as a BCA and the information contained here serves for its registration and commercialization in Europe.

  5. Use of active extracts of poplar buds against Penicillium italicum and possible modes of action.

    PubMed

    Yang, Shuzhen; Liu, Limei; Li, Dongmei; Xia, Huan; Su, Xiaojun; Peng, Litao; Pan, Siyi

    2016-04-01

    Antifungal components, from poplar buds active fraction (PBAF) against Penicillium italicum, the causal agent of blue mold in citrus fruits, were identified and possible action modes were investigated. Pinocembrin, chrysin and galangin were determined as active components in PBAF, using HPLC and HPLC-MS analysis. The antifungal activity is stable at temperatures ranging from 4 °C to 100 °C and pH levels ranging from 4 to 8. In the presence of PBAF, the hyphae become shriveled, wrinkled and the cell membrane became seriously disrupted. Further investigation on cell permeability, nucleic acid content and alkaline phosphatase suggest that the cell membrane might be the target. Mycelial oxygen consumption and the respiration-related enzymatic activity of succinate dehydrogenase, malate dehydrogenase and ATPase were all inhibited by PBAF. We propose that PBAF is a potentially useful alternative for blue mold control and may act against P. italicum by interfering with respiration and disrupting the cell membrane.

  6. Travel-related disseminated Penicillium marneffei infection in a renal transplant patient.

    PubMed

    Hart, J; Dyer, J R; Clark, B M; McLellan, D G; Perera, S; Ferrari, P

    2012-08-01

    Penicillium marneffei is a thermally dimorphic fungus that causes severe human immunodeficiency virus-related opportunistic infection in endemic areas of Southeast Asia and has rarely been reported in solid organ transplant (SOT) recipients. We report here the case of an Australian renal transplant patient who presented with disseminated P. marneffei infection shortly after a 10-day holiday to Vietnam, and review all previously published cases of penicilliosis associated with renal transplantation. This is the first reported case, to our knowledge, of P. marneffei infection in an SOT recipient acquired during travel to an endemic country, and highlights the importance of an accurate travel history when opportunistic infection is suspected, as well as giving appropriate health advice to transplant patients who travel.

  7. Morphology engineering of Penicillium chrysogenum by RNA silencing of chitin synthase gene.

    PubMed

    Liu, Hui; Wang, Peng; Gong, Guohong; Wang, Li; Zhao, Genhai; Zheng, Zhiming

    2013-03-01

    Chitin synthases, that catalyze the formation of chitin the major component of cell walls in most filamentous fungi, play crucial roles in the growth and morphogenesis. To investigate the roles of chitin synthase in Penicillium chrysogenum, we developed an RNAi system to silence the class III chitin synthase gene chs4. After transformation, mutants had a slow growth rate and shorter but highly branched hyphae. All transformants either were unable to form conidia or could form only a few. Changes in chs4 expression could lead to a completely different morphology and eventually cause distinct penicillin yields. In particular, the yield of one transformant was 41 % higher than that of the original strain.

  8. Aminoacyl-coenzyme A synthesis catalyzed by a CoA ligase from Penicillium chrysogenum.

    PubMed

    Koetsier, Martijn J; Jekel, Peter A; Wijma, Hein J; Bovenberg, Roel A L; Janssen, Dick B

    2011-03-23

    Coenzyme A ligases play an important role in metabolism by catalyzing the activation of carboxylic acids. In this study we describe the synthesis of aminoacyl-coenzyme As (CoAs) catalyzed by a CoA ligase from Penicillium chrysogenum. The enzyme accepted medium-chain length fatty acids as the best substrates, but the proteinogenic amino acids L-phenylalanine and L-tyrosine, as well as the non-proteinogenic amino acids D-phenylalanine, D-tyrosine and (R)- and (S)-β-phenylalanine were also accepted. Of these amino acids, the highest activity was found for (R)-β-phenylalanine, forming (R)-β-phenylalanyl-CoA. Homology modeling suggested that alanine 312 is part of the active site cavity, and mutagenesis (A312G) yielded a variant that has an enhanced catalytic efficiency with β-phenylalanines and D-α-phenylalanine.

  9. Promotion of extracellular lignocellulolytic enzymes production by restraining the intracellular β-glucosidase in Penicillium decumbens.

    PubMed

    Chen, Mei; Qin, Yuqi; Cao, Qing; Liu, Guodong; Li, Jie; Li, Zhonghai; Zhao, Jian; Qu, Yinbo

    2013-06-01

    In this study, the functions of β-glucosidases in regulation of the lignocellulolytic enzymes production in Penicillium decumbens 114-2 were investigated. The major extracellular β-glucosidase gene bgl1 and the major intracellular β-glucosidase gene bgl2 were deleted in P. decumbens 114-2 respectively. In Δbgl2, the production of extracellular lignocellulolytic enzymes (including endoglucanases, cellobiohydrolases and xylanases) on insoluble cellulose was significantly promoted, while in Δbgl1 there was no any difference compared with that of 114-2. The enhancement of the production of lignocellulolytic enzymes in Δbgl2 was likely attributed to the accumulation of intracellular cellobiose. Induction experiment in Δbgl1Δbgl2 showed that cellobiose was an inducer of lignocellulolytic enzymes expression in P. decumbens 114-2, and the induction was unrelated to the formation, if any, of gentiobiose or sophorose from cellobiose.

  10. Secondary Metabolism in Penicillium expansum: Emphasis on Recent Advances in Patulin Research.

    PubMed

    Tannous, Joanna; Keller, Nancy P; Atoui, Ali; El Khoury, André; Lteif, Roger; Oswald, Isabelle P; Puel, Olivier

    2017-03-31

    The plant pathogenic fungus Penicillium expansum is a major concern of the global food industry due to its wide occurrence and ability to produce various mycotoxins, of which the most significant is patulin. Relatively less highlighted in the literature, in comparison with the other food-borne mycotoxins, patulin is one of the main factors in economic losses of vegetables and fruits. Otherwise, patulin is a health hazard which results in both short-term and long-term risks. This review includes knowledge on the biosynthetic mechanisms used for secondary metabolite production in P. expansum, with special emphasis on patulin biosynthesis. The abiotic factors triggering the production of patulin and the strategies developed to reduce or prevent the contamination by this mycotoxin are comprehensively discussed. The database presented in this review would be useful for the prioritization and development of future research.

  11. Oxidative damage involves in the inhibitory effect of nitric oxide on spore germination of Penicillium expansum.

    PubMed

    Lai, Tongfei; Li, Boqiang; Qin, Guozheng; Tian, Shiping

    2011-01-01

    The effects of nitric oxide (NO) on spore germination of Penicillium expansum were investigated and a possible mechanism was evaluated. The results indicated that NO released by sodium nitroprusside (SNP) significantly suppressed fungal growth. With the use of an oxidant sensitive probe and Western blot analysis, an increased level of intracellular reactive oxygen species (ROS) and enhanced carbonylation damage were detected in spores of P. expansum under NO stress. Exogenous superoxide dismutase (SOD) and ascorbic acid (Vc) could increase the resistance of the spore to the inhibitory effect of NO. The activities of SOD and catalase (CAT), as well as ATP content in spores under NO stress were also lower than those in the control. We suggest that NO in high concentration induces the generation of ROS which subsequently causes severe oxidative damage to proteins crucial to the process of spore germination of P. expansum.

  12. Bergamotane Sesquiterpenes with Alpha-Glucosidase Inhibitory Activity from the Plant Pathogenic Fungus Penicillium expansum.

    PubMed

    Ying, You-Min; Fang, Cheng-An; Yao, Feng-Qi; Yu, Yuan; Shen, Ying; Hou, Zhuo-Ni; Wang, Zhen; Zhang, Wei; Shan, Wei-Guang; Zhan, Zha-Jun

    2017-01-01

    Two new bergamotane sesquiterpene lactones, named expansolides C and D (1 and 2), together with two known compounds expansolides A and B (3 and 4), were isolated from the plant pathogenic fungus Penicillium expansum ACCC37275. The structures of the new compounds were established by detailed analyses of the spectroscopic data, especially 1D-, 2D-NMR, and HR-ESI-MS. In an in vitro bioassay, the epimeric mixture of expansolides C and D (1 and 2) (in a ratio of 2:1 at the temprature of the bioassay) exhibited more potent α-glucosidase inhibitory activity (IC50 =0.50 ± 0.02 mm) as compared with the positive control acarbose (IC50 = 1.90 ± 0.05 mm). To the best of our knowledge, it was the first report on the α-glucosidase inhibitory activity of bergamotane sesquiterpenes.

  13. Molecular cloning and characterization of the glyceraldehyde-3-phosphate dehydrogenase gene from Penicillium expansum PE-12.

    PubMed

    Zhang, T; Qi, Z; Yu, Q S; Tang, K X

    2013-07-15

    Penicillium expansum produces large amounts of lipase, which is widely used in laundry detergent and leather industry. We isolated the glyceraldehyde-3-phosphate dehydrogenase gene (PeGPD) from P. expansum PE-12 through reverse transcriptase PCR and 5'-3' rapid amplification of cDNA ends (RACE-PCR). The gene is 1266 bp long, including an ORF of 1014 bp, encoding a polypeptide chain of 337 amino acids. A phylogenetic tree based on GPD proteins showed that P. expansum is close to Aspergillus species, but comparatively distant from P. marneffei. Southern blot results revealed a single copy of PeGPD, and expression analysis gave evidence of high expression levels. PeGPD genes have potential for genetic engineering of P. expansum for industrial lipase production.

  14. Residue Val237 is critical for the enantioselectivity of Penicillium expansum lipase.

    PubMed

    Tang, Lianghua; Su, Min; Chi, Liying; Zhang, Junling; Zhang, Huihui; Zhu, Ling

    2014-03-01

    The shape of the hydrophobic tunnel leading to the active site of Penicillium expansum lipase (PEL) was redesigned by single-point mutations, in order to better understand enzyme enantioselectivity towards naproxen. A variant with a valine-to-glycine substitution at residue 237 exhibited almost no enantioselectivity (E = 1.1) compared with that (E = 104) of wild-type PEL. The function of the residue, Val237, in the hydrophobic tunnel was further analyzed by site-directed mutagenesis. For each of these variants a significant decrease of enantioselectivity (E < 7) was observed compared with that of wild-type enzyme. Further docking result showed that Val237 plays the most important role in stabilizing the correct orientation of (R)-naproxen. Overall, these results indicate that the residue Val237 is the key amino acid residue maintaining the enantioselectivity of the lipase.

  15. Alkaloids with Cardiovascular Effects from the Marine-Derived Fungus Penicillium expansum Y32.

    PubMed

    Fan, Ya-Qin; Li, Pei-Hai; Chao, Ya-Xi; Chen, Hao; Du, Ning; He, Qiu-Xia; Liu, Ke-Chun

    2015-10-22

    Three new alkaloids (1, 4 and 8), together with nine known analogues (2, 3, 5-7, and 9-12), were isolated from the marine-derived fungus Penicillium expansum Y32. Their structures including the absolute configurations were elucidated by spectroscopic and Mosher's and Marfey's methods, along with quantum electronic circular dichroism (ECD) calculations. Each of the compounds was evaluated for cardiovascular effects in a live zebrafish model. All of the compounds showed a significant mitigative effect on bradycardia caused by astemizole (ASM) in the heart rate experiments. Compounds 4-6 and 8-12 exhibited potent vasculogenetic activity in vasculogenesis experiments. This is the first study to report that these types of compounds show cardiovascular effects in zebrafish. The results suggest that these compounds could be promising candidates for cardiovascular disease lead compounds.

  16. Antimicrobial effects of ionizing radiation on artificially and naturally contaminated cacao beans. [Aspergillus flavus; Penicillium citrinum

    SciTech Connect

    Restaino, L.; Myron, J.J.J.; Lenovich, L.M.; Bills, S.; Tscherneff, K.

    1984-04-01

    With an initial microbial level of ca. 10/sup 7/ microorganisms per g of Ivory Coast cacao beans, 5 kGy of gamma radiation from a Co/sup 60/ source under an atmosphere of air reduced the microflora per g by 2.49 and 3.03 logs at temperatures of 35 and 50/sup 0/C, respectively. Bahia cacao beans were artificially contaminated with dried spores of Aspergillus flavus and Penicillium citrinum, giving initial fungal levels of 1.9 x 10/sup 4/ and 1.4 x 10/sup 3/ spores per g of whole Bahia cacao beans, respectively. The average D/sub 10/ values for A. flavus and P. citrinum spores on Bahia cacao beans were 0.66 and 0.88 kGy, respectively. 12 references.

  17. Influence of Different Nanomaterials on Growth and Mycotoxin Production of Penicillium verrucosum

    PubMed Central

    Kotzybik, Kathrin; Gräf, Volker; Kugler, Lena; Stoll, Dominic A.; Greiner, Ralf; Geisen, Rolf; Schmidt-Heydt, Markus

    2016-01-01

    Nanoparticles are ubiquitous in the environment. They originate from anthropogenic or natural sources or they are intentionally produced for different purposes. There exist manifold applications of nanoparticles in modern life leading unavoidably to a confrontation and interaction between nanomaterial and living organisms. Based on their wide distribution tending to increase steadily, the influence of particles based on silica and silver, exhibiting nominal sizes between 0.65 nm and 200 nm, on the physiology of the mycotoxigenic filamentous fungus Penicillium verrucosum was analyzed. The applied concentration and time-point, the size and the chemical composition of the particles was shown to have a strong influence on growth and mycotoxin biosynthesis. On microscopic scale it could be shown that silver nanoparticles attach to the mycelial surface. Moreover, silver nanoparticles with 0.65 nm and 5 nm in size were shown to internalize within the cell, form agglomerates in the cytoplasm and associate to cell organelles. PMID:26974550

  18. Fleming's penicillin producing strain is not Penicillium chrysogenum but P. rubens.

    PubMed

    Houbraken, Jos; Frisvad, Jens C; Samson, Robert A

    2011-06-01

    Penicillium chrysogenum is a commonly occurring mould in indoor environments and foods, and has gained much attention for its use in the production of the antibiotic penicillin. Phylogenetic analysis of the most important penicillin producing P. chrysogenum isolates revealed the presence of two highly supported clades, and we show here that these two clades represent two species, P. chrysogenum and P. rubens. These species are phenotypically similar, but extrolite analysis shows that P. chrysogenum produces secalonic acid D and F and/or a metabolite related to lumpidin, while P. rubens does not produce these metabolites. Fleming's original penicillin producing strain and the full genome sequenced strain of P. chrysogenum are re-identified as P. rubens. Furthermore, the well-known claim that Alexander Fleming misidentified the original penicillin producing strain as P. rubrum is discussed.

  19. Acute Abdomen Due to Penicillium marneffei: An Indicator of HIV Infection in Manipur State.

    PubMed

    Ghalige, Hemanth Sureshwara; Sahoo, Biswajeet; Sharma, Sanjeeb; Devi, Khuraijam Ranjana; Singh Th, Sudhir Chandra

    2014-09-01

    Opportunistic infection in HIV disease often present to clinicians in an atypical manner testing clinical acumen. Here, we report a case of Penicilliosis marneffei (PM) infection presenting to surgical emergency as acute abdomen with undiagnosed HIV status in advanced AIDS, chief complaints being prolonged fever and diffuse abdominal pain. Radiologic imaging showed non-specific mesenteric and retroperitoneal lymphadenopathy. Fine needle aspiration cytology (FNAC) of the lymph node was done and subjected to direct microscopy, gram staining and culture on Sabouraud's dextrose agar (SDA) which showed Penicillium marneffei. He was then treated with intravenous amphotericin. This case is reported for its rarity and unusual presentation to sensitise clinicians and microbiologists to consider PM as an aetiology in acute abdomen in high risk individuals, more so, in patients from north-east India.

  20. Combined effects of selected Penicillium mycotoxins on in vitro proliferation of porcine lymphocytes.

    PubMed

    Bernhoft, Aksel; Keblys, Modestas; Morrison, Ellen; Larsen, Hans Jørgen S; Flåøyen, Arne

    2004-11-01

    The in vitro effect of combinations of the Penicillium mycotoxins citrinin (CIT), cyclopiazonic acid (CPA), ochratoxin A (OTA), patulin (PAT), penicillic acid (PIA) and roquefortine C (RQC) on mitogen induced lymphocyte proliferation was determined using purified lymphocytes from six piglets. Dose-response curves for each mycotoxin and mycotoxin combinations were generated. The combined effects of toxin pairs based on IC20 were illustrated in isobole diagrams and statistically calculated. OTA and CIT elicited a synergistic effect. Four toxin pairs elicited additive effects, four pairs less-than-additive effects and six pairs independent effects. Thus, the majority of toxin pairs tested produced lower combined effects than an additive effect. The results indicate that the sum effect of all toxins is less than that from the summation of concentrations of the individual compounds, adjusted for differences in potencies.

  1. A Sterol and Spiroditerpenoids from a Penicillium sp. Isolated from a Deep Sea Sediment Sample

    PubMed Central

    Li, Yan; Ye, Dezan; Shao, Zongze; Cui, Chengbin; Che, Yongsheng

    2012-01-01

    A new polyoxygenated sterol, sterolic acid (1), three new breviane spiroditerpenoids, breviones I–K (2–4), and the known breviones (5–8), were isolated from the crude extract of a Penicillium sp. obtained from a deep sea sediment sample that was collected at a depth of 5115 m. The structures of 1–4 were elucidated primarily by NMR experiments, and 1 was further confirmed by X-ray crystallography. The absolute configurations of 2 and 3 were deduced by comparison of their CD spectra with those of the model compounds. Compounds 2 and 5 showed significant cytotoxicity against MCF-7 cells, which is comparable to the positive control cisplatin. PMID:22412815

  2. Citral exerts its antifungal activity against Penicillium digitatum by affecting the mitochondrial morphology and function.

    PubMed

    Zheng, Shiju; Jing, Guoxing; Wang, Xiao; Ouyang, Qiuli; Jia, Lei; Tao, Nengguo

    2015-07-01

    This work investigated the effect of citral on the mitochondrial morphology and function of Penicillium digitatum. Citral at concentrations of 2.0 or 4.0 μL/mL strongly damaged mitochondria of test pathogen by causing the loss of matrix and increase of irregular mitochondria. The deformation extent of the mitochondria of P. digitatum enhanced with increasing concentrations of citral, as evidenced by a decrease in intracellular ATP content and an increase in extracellular ATP content of P. digitatum cells. Oxygen consumption showed that citral resulted in an inhibition in the tricarboxylic acid cycle (TCA) pathway of P. digitatum cells, induced a decrease in activities of citrate synthetase, isocitrate dehydrogenase, α-ketoglutarate dehydrogenase, succinodehydrogenase and the content of citric acid, while enhancing the activity of malic dehydrogenase in P. digitatum cells. Our present results indicated that citral could damage the mitochondrial membrane permeability and disrupt the TCA pathway of P. digitatum.

  3. Six New Polyketide Decalin Compounds from Mangrove Endophytic Fungus Penicillium aurantiogriseum 328#

    PubMed Central

    Ma, Yanhong; Li, Jing; Huang, Meixiang; Liu, Lan; Wang, Jun; Lin, Yongcheng

    2015-01-01

    Six new compounds with polyketide decalin ring, peaurantiogriseols A–F (1–6), along with two known compounds, aspermytin A (7), 1-propanone,3-hydroxy-1-(1,2,4a,5,6,7,8,8a-octahydro-2,5-dihydroxy-1,2,6-trimethyl-1-naphthalenyl) (8), were isolated from the fermentation products of mangrove endophytic fungus Penicillium aurantiogriseum 328#. Their structures were elucidated based on their structure analysis. The absolute configurations of compounds 1 and 2 were determined by 1H NMR analysis of their Mosher esters; the absolute configurations of 3–6 were determined by using theoretical calculations of electronic circular dichroism (ECD). Compounds 1–8 showed low inhibitory activity against human aldose reductase, no activity of inducing neurite outgrowth, nor antimicrobial activity. PMID:26473887

  4. Production of xylanase and protease by Penicillium janthinellum CRC 87M-115 from different agricultural wastes.

    PubMed

    Oliveira, Luciana A; Porto, Ana L F; Tambourgi, Elias B

    2006-04-01

    Five agricultural wastes were evaluated in submerged fermentation for xylanolytic enzymes production by Penicillium janthinellum. The wastes were hydrolyzed in acid medium and the liquid fraction was used for cultivation. Corn cob (55.3 U/mL) and oat husk (54.8 U/mL) were the best inducers of xylanase. Sugar cane bagasse (23.0 U/mL) and corn husk (23.8 U/mL) were moderately good, while cassava peel was negligible. Protease production was very low in all agro-industrial residues. The maximum biomass yields were 1.30 and 1.17 g/L for cassava peel and corn husk after 180 h, respectively. Xylanolytic activity showed a cell growth associated profile.

  5. Inhibition of Aspergillus spp. and Penicillium spp. by fatty acids and their monoglycerides.

    PubMed

    Altieri, Clelia; Cardillo, Daniela; Bevilacqua, Antonio; Sinigaglia, Milena

    2007-05-01

    The antifungal activity of three fatty acids (lauric, myristic, and palmitic acids) and their monoglycerides (monolaurin, monomyristic acid, and palmitin, respectively) against Aspergillus and Penicillium species in a model system was investigated. Data were modeled through a reparameterized Gompertz equation. The maximum colony diameter attained within the experimental time (30 days), the maximal radial growth rate, the lag time (i.e., the number of days before the beginning of radial fungal growth), and the minimum detection time (MDT; the number of days needed to attain 1 cm colony diameter) were evaluated. Fatty acids and their monoglycerides inhibited mold growth by increasing MDT and lag times. The effectiveness of the active compounds seemed to be strain and genus dependent. Palmitic acid was the most effective chemical against aspergilli, whereas penicilli were strongly inhibited by myristic acid. Aspergilli also were more susceptible to fatty acids than were penicilli, as indicated by the longer MDT.

  6. Bioleaching of rare earth and radioactive elements from red mud using Penicillium tricolor RM-10.

    PubMed

    Qu, Yang; Lian, Bin

    2013-05-01

    The aim of this work is to investigate biological leaching of rare earth elements (REEs) and radioactive elements from red mud, and to evaluate the radioactivity of the bioleached red mud used for construction materials. A filamentous, acid-producing fungi named RM-10, identified as Penicillium tricolor, is isolated from red mud. In our bioleaching experiments by using RM-10, a total concentration of 2% (w/v) red mud under one-step bioleaching process was generally found to give the maximum leaching ratios of the REEs and radioactive elements. However, the highest extraction yields are achieved under two-step bioleaching process at 10% (w/v) pulp density. At pulp densities of 2% and 5% (w/v), red mud processed under both one- and two-step bioleaching can meet the radioactivity regulations in China.

  7. Efficient production and evaluation of lignocellulolytic enzymes using a constitutive protein expression system in Penicillium oxalicum.

    PubMed

    Hu, Yibo; Xue, Haizhao; Liu, Guodong; Song, Xin; Qu, Yinbo

    2015-06-01

    Native lignocellulolytic enzyme systems secreted by filamentous fungi can be further optimized by protein engineering or supplementation of exogenous enzyme components. We developed a protein production and evaluation system in cellulase-producing fungus Penicillium oxalicum. First, by deleting the major amylase gene amy15A, a strain Δ15A producing few extracellular proteins on starch was constructed. Then, three lignocellulolytic enzymes (BGL4, Xyn10B, and Cel12A) with originally low expression levels were successfully expressed with selected constitutive promoters in strain Δ15A. BGL4 and Cel12A overexpression resulted in increased specific filter paper activity (FPA), while the overexpression of Xyn10B improved volumetric FPA but not specific FPA. By switching the culture medium, this platform is convenient to produce originally low-expressed lignocellulolytic enzymes in relatively high purities on starch and to evaluate the effect of their supplementation on the performance of a complex cellulase system on cellulose.

  8. Brevianamides and Mycophenolic Acid Derivatives from the Deep-Sea-Derived Fungus Penicillium brevicompactum DFFSCS025

    PubMed Central

    Xu, Xinya; Zhang, Xiaoyong; Nong, Xuhua; Wang, Jie; Qi, Shuhua

    2017-01-01

    Four new compounds (1–4), including two brevianamides and two mycochromenic acid derivatives along with six known compounds were isolated from the deep-sea-derived fungus Penicillium brevicompactum DFFSCS025. Their structures were elucidated by spectroscopic analysis. Moreover, the absolute configurations of 1 and 2 were determined by quantum chemical calculations of the electronic circular dichroism (ECD) spectra. Compound 9 showed moderate cytotoxicity against human colon cancer HCT116 cell line with IC50 value of 15.6 μM. In addition, 3 and 5 had significant antifouling activity against Bugula neritina larval settlement with EC50 values of 13.7 and 22.6 μM, respectively. The NMR data of 6, 8, and 9 were assigned for the first time. PMID:28218640

  9. Evaluation of chitosans and Pichia guillermondii as growth inhibitors of Penicillium digitatum.

    PubMed

    Pacheco, Neith; Larralde-Corona, C Patricia; Sepulveda, Jose; Trombotto, Stéphan; Domard, Alain; Shirai, Keiko

    2008-07-01

    Chitosans were obtained by room-temperature-homogeneous-deacetylation (RTHD) and freeze-pump-out-thaw-heterogeneous-deacetylation (FPT) from chitins purified from fermentations. Commercial chitosan was deacetylated by three-FPT-cycles. Chitosans and Pichia guillermondii were evaluated on the growth of Penicillium digitatum. Medium molecular weight (M(W)) chitosans displayed higher inhibitory activity against the yeast than low M(W) biopolymers. Chitosans with low degree of acetylation (DA) were inhibitory for yeast and mould. Therefore, a low M(W) and high DA chitosan was selected for use against moulds combined with yeasts. Biopolymer and yeasts presented an additive effect, since chitosans were effective to delay spore germination, whereas yeast decreased apical fungal growth.

  10. A kinetic study of textile dyeing wastewater degradation by Penicillium chrysogenum.

    PubMed

    Durruty, Ignacio; Fasce, Diana; González, Jorge Froilán; Wolski, Erika Alejandra

    2015-06-01

    The potential of Penicillium chrysogenum to decolorize azo dyes and a real industrial textile wastewater was studied. P. chrysogenum was able to decolorize and degrade three azo dyes (200 mg L(-1)), either independently or in a mixture of them, using glucose as a carbon source. A kinetic model for degradation was developed and it allowed predicting the degradation kinetics of the mixture of the three azo dyes. In addition, P. chrysogenum was able to decolorize real industrial wastewater. The kinetic model proposed was also able to predict the decolorization of the real wastewater. The calibration of the proposed model makes it a useful tool for future wastewater facilities' design and for practical applications.

  11. Penicillium expansum Link strain for a biometallurgical method to recover REEs from WEEE.

    PubMed

    Di Piazza, Simone; Cecchi, Grazia; Cardinale, Anna Maria; Carbone, Cristina; Mariotti, Mauro Giorgio; Giovine, Marco; Zotti, Mirca

    2017-02-01

    Due to the wide range of applications in high-tech solutions, Rare Earth Elements (REEs) have become object of great interest. In the last years several studies regarding technologies for REE extraction from secondary resources have been carried out. In particular biotechnologies, which use tolerant and accumulator microorganisms to recover and recycle precious metals, are replacing traditional methods. This paper describes an original biometallurgical method to recover REEs from waste electrical and electronic equipment (WEEE) by using a strain of Penicillium expansum Link isolated from an ecotoxic metal contaminated site. The resulting product is a high concentrated solution of Lanthanum (up to 390ppm) and Terbium (up to 1520ppm) obtained from WEEE. Under this perspective, the proposed protocol can be considered a method of recycling exploiting biometallurgy. Finally, the process is the subject of the Italian patent application n. 102015000041404 submitted by the University of Genoa.

  12. Primary structure of a guanyl-specific ribonuclease from the fungus Penicillium brevicompactum

    SciTech Connect

    Kulikov, V.A.; Shlyapnikov, S.V.; Yakovlev, G.I.

    1986-01-01

    By the automatic Edman degradation of the intact S-carboxymethylated protein and a mixture of the products of its proteolytic cleavage at Arg, Lys, and Glu residues, together with results on the kinetics of the proteolysis of the protein under the action of carboxypeptidase Y, the primary structure of the extracellular guanyl-specific RNase of the fungus Penicillium brevicompactum has been determined. The RNase contains 102 amino acid residues: 7 Asp, 7 Asn, 9 Thr, 11 Ser, 4 Glu, 1 Gln, 4 Pro, 10 Gly, 11 Ala, 4 Cys, 7 Val, 4 Ile, 3 Leu, 9 Tyr, 5 Phe, 2 Lys, 3 His, 1 Arg (M/sub r/ 10,801). It has been established that four hemicystine residues of the P. compactum RNase form, in pairs, two disulfide bonds

  13. Role of intracellular free calcium in killing Penicillium marneffei within human macrophages.

    PubMed

    Chen, Renqiong; Ji, Guangquan; Ma, Tuan; Huang, Xiaowen; Ren, Hong; Xi, Liyan

    2015-01-01

    Increases in cytosolic Ca(2+) concentration ([Ca(2+)]c) promote phagocyte antimicrobial responses. Here, we investigated macrophages stimulated by Penicillium marneffei (P. marneffei). [Ca(2+)]c was determined in macrophages loaded with the fluorescent calcium probe Fura 2/AM as they were stimulated by P. marneffei. We found that P. marneffei induced an increase in [Ca(2+)]c in human macrophages. Further, increased [Ca(2+)]c with the ionophore A23187 promoted phagosomal acidification and maturation and reduced intracellular replication of P. marneffei in P. marneffei-infected human macrophages, whereas decreased [Ca(2+)]c with the chelation MAPTAM decreased TNF-α production, inhibited phagosomal acidification and maturation and increased intracellular replication of P. marneffei. These data indicate that Ca(2+) signaling may play an important role in controlling the replication of P. marneffei within macrophages.

  14. Quantitative clarification of inactivation mechanism of Penicillium digitatum spores treated with neutral oxygen radicals

    NASA Astrophysics Data System (ADS)

    Hashizume, Hiroshi; Ohta, Takayuki; Takeda, Keigo; Ishikawa, Kenji; Hori, Masaru; Ito, Masafumi

    2015-01-01

    We have quantitatively investigated the oxidative inactivation process of Penicillium digitatum spores including intracellular nanostructural changes through neutral oxygen species with a flux-defined atmospheric-pressure oxygen radical source, using fluorescent confocal-laser microscopy and transmission electron microscopy (TEM). The results suggest that neutral oxygen species, particularly ground-state atomic oxygen [O(3Pj)], which is an effective species for inactivating P. digitatum spores, inhibit the function of the cell membrane of spores without causing major superficial morphological changes at a low O(3Pj) dose of ˜2.1 × 1019 cm-2 under an O(3Pj) flux of 2.3 × 1017 cm-2 s-1, following the oxidation of intracellular organelles up to an O(3Pj) dose of ˜1.0 × 1020 cm-2. Finally, intracellular nanostructures are degraded by excess oxygen radicals over an O(3Pj) dose of ˜1.0 × 1020 cm-2.

  15. Studies on cellulase production by a mutant - Penicillium funiculosum UV-49

    SciTech Connect

    Joglekar, A.V.; Karanth, N.G.

    1984-01-01

    Rapidly depleting reserves and the rising cost of petroleum has focused worldwide attention on the development of alternate sources of energy. One of the most promising potential solutions to this problem is utilization of cellulosic materials through bioconversion. In search of hypercelluloytic microorganisms, ultraviolet irradiation carried out with Penicillium funiculosum has yielded a superior mutant. The investigations reported in this article are shake flask studies on some important nutritional requirements of the mutant, namely, nitrogen source, carbon source, and inducers. The mutant shows an ability to metabolize inorganic nitrogen sources like urea and sodium nitrate both for growth and enzyme production. A comparison of the long-term saccharification ability and the utilization efficiency of the mutant enzyme with those reported in the literature is also carried out, showing the superior performance of the mutant enzyme.

  16. Penicillium roqueforti: a multifunctional cell factory of high value-added molecules.

    PubMed

    Mioso, R; Toledo Marante, F J; Herrera Bravo de Laguna, I

    2015-04-01

    This is a comprehensive review, with 114 references, of the chemical diversity found in the fungus Penicillium roqueforti. Secondary metabolites of an alkaloidal nature are described, for example, ergot alkaloids such as festuclavine, isofumigaclavines A and B, and diketopiperazine alkaloids such as roquefortines A-D, which are derived from imidazole. Other metabolites are marcfortines A-C, PR-toxin, eremofortines A-E, mycophenolic and penicillic acids, and some γ-lactones. Also, recent developments related to the structural characteristics of botryodiplodin and andrastin are studied-the latter has anticancer properties. Finally, we discuss the enzymes of P. roqueforti, which can participate in the biotechnological production of high value-added molecules, as well as the use of secondary metabolite profiles for taxonomic purposes.

  17. Untargeted Metabolic Profiling of Winery-Derived Biomass Waste Degradation by Penicillium chrysogenum.

    PubMed

    Karpe, Avinash V; Beale, David J; Godhani, Nainesh B; Morrison, Paul D; Harding, Ian H; Palombo, Enzo A

    2015-12-16

    Winery-derived biomass waste was degraded by Penicillium chrysogenum under solid state fermentation over 8 days in a (2)H2O-supplemented medium. Multivariate statistical analysis of the gas chromatography-mass spectrometry (GC-MS) data resulted in the identification of 94 significant metabolites, within 28 different metabolic pathways. The majority of biomass sugars were utilized by day 4 to yield products such as sugars, fatty acids, isoprenoids, and amino acids. The fungus was observed to metabolize xylose to xylitol, an intermediate of ethanol production. However, enzyme inhibition and autolysis were observed from day 6, indicating 5 days as the optimal time for fermentation. P. chrysogenum displayed metabolism of pentoses (to alcohols) and degraded tannins and lignins, properties that are lacking in other biomass-degrading ascomycetes. Rapid fermentation (3-5 days) may not only increase the pentose metabolizing efficiency but also increase the yield of medicinally important metabolites, such as syringate.

  18. [Synthesis and identification of penicillic-acid antigens from Penicillium cyclopium].

    PubMed

    Lei, Hongyu; Yuan, Hui; Wu, Jing; Yuan, Liyun; Wen, Lixin; Ni, Hengjia

    2008-05-01

    To establish a new immune assay for Penicillic Acid (PA) from Penicillium cyclopium, we studied the synthesis of conjugated complete antigens for penicillic acid. PA was conjugated to bovine serum album (BSA) and ovalbumin (OVA) by 1-ethyl-3-(3-dimethyl-aminopropyl) carbodiimide hydrochloride (EDC). The artificial antigens PA-BSA and PA-OVA were identified by ultraviolet spectrometric scanning, SDS-PAGE and immunization. Results showed that the absorption peak of conjugation were different from that of the carrier protein alone and of the PA. The conjugated ratio of PA and BSA was 23.2:1 and that of PA and OVA was 10.4:1. Balb/c mice were immunized by the artificial antigen of PA-BSA, with PA-OVA as coating antigen. The average titer of antiserums was more than 12 800 by indirect ELISA. The obtained antigens offered a basis for developing immunoassay method.

  19. Identification of the viridicatumtoxin and griseofulvin gene clusters from Penicillium aethiopicum.

    PubMed

    Chooi, Yit-Heng; Cacho, Ralph; Tang, Yi

    2010-05-28

    Penicillium aethiopicum produces two structurally interesting and biologically active polyketides: the tetracycline-like viridicatumtoxin 1 and the classic antifungal agent griseofulvin 2. Here, we report the concurrent discovery of the two corresponding biosynthetic gene clusters (vrt and gsf) by 454 shotgun sequencing. Gene deletions confirmed that two nonreducing PKSs (NRPKSs), vrtA and gsfA, are required for the biosynthesis of 1 and 2, respectively. Both PKSs share similar domain architectures and lack a C-terminal thioesterase domain. We identified gsfI as the chlorinase involved in the biosynthesis of 2, because deletion of gsfI resulted in the accumulation of decholorogriseofulvin 3. Comparative analysis with the P. chrysogenum genome revealed that both clusters are embedded within conserved syntenic regions of P. aethiopicum chromosomes. Discovery of the vrt and gsf clusters provided the basis for genetic and biochemical studies of the pathways.

  20. Characterization of novel thermostable polygalacturonases from Penicillium brasilianum and Aspergillus niger.

    PubMed

    Zeni, Jamile; Pili, Jonaina; Cence, Karine; Toniazzo, Geciane; Treichel, Helen; Valduga, Eunice

    2015-12-01

    The aim of this research was the partial characterization of polygalacturonase (PG) extracts produced by a newly isolated Penicillium brasilianum and Aspergillus niger in submerged fermentation. The partial characterization of the crude enzymatic extracts showed optimum activity at pH 5.5 and 37 °C for both extracts. The results of temperature stability showed that PG from both microorganisms were more stable at 55 °C. However, the enzyme obtained by P. brasilianum presents a half-life time (t 1/2 = 693.10 h), about one order of magnitude higher than those observed in for A. niger at 55 °C. In terms of pH stability, the PG produced by P. brasilianum presented higher stability at pH 4.0 and 5.0, while the PG from A. niger showed higher stability at pH 5.0.

  1. [Exometabolites of the Fungal Isolates (Genus Penicillium, Section Chrysogena) from Low-Temperature Ecotopes].

    PubMed

    Kozlovskii, A G; Antipova, T V; Zhelifonova, V P; Baskunov, B P; Kochkina, G A; Ozerskaya, S M

    2016-01-01

    Exometabolites of 22 strains of the genus Penicillium, section Chrysogena isolated from low-temperature ecotopes of various geographical regions were analyzed. The ecotopes included permafrost deposits, frozen volcanic ash, a fossil horse, cryopeg, and water from an Antarctic lake. The studied strains were found to contain exometabolites belonging to the groups of penicillins (penicillin G), chrysogins (chrysogin, 3-acetylquinazolone-4, 2-pyruvoyl aminobenzamide, 2-hydroxypropionyl amunobenzamide, and questiomycin A), roquefortines (3,12-dihydroroquefortine, roquefortine, glandicolines A and B, and meleagrine), xanthocillins (xanthocillin X), and simple tryptophan derivatives (N-acetyltriptamine and indoleacetic acid). In five P. chrysogenum strains and three P. nalgiovense strains a correlation was found between exometabolite spectra and morphological characteristics of the cultures isolated from modern ecotopes. For other strains species identification was based on morphological features, due to the absence of biosynthesis of penicillin G, on of the major chemotaxonomic markers for these species.

  2. Application of nested PCR to detect Penicillium marneffei in serum samples.

    PubMed

    Pongpom, Monsicha; Sirisanthana, Thira; Vanittanakom, Nongnuch

    2009-01-01

    We previously reported a nested PCR assay for specific identification of 18S ribosomal DNA of Penicillium marneffei. In this study, the assay was used to detect the DNA of P. marneffei in serum samples. Sensitivity of the test was 4 pg/microl and 0.4 fg/microl when the cycle numbers used for nested reactions were 15 and 30, respectively. Twenty four out of 35 sera (68.6%) collected from patients with culture confirmed penicilliosis marneffei were positive, while normal healthy and non-P. marneffei infected HIV-positive sera were negative. The results suggested that the assay could be applied for the diagnosis of infections due to P. marneffei.

  3. Polymerase chain reaction (PCR) identification of Penicillium brevicompactum, a grape contaminant and mycophenolic acid producer.

    PubMed

    Patiño, B; Medina, A; Doménech, M; González-Jaén, M T; Jiménez, M; Vázquez, C

    2007-02-01

    Penicillium brevicompactum is a ubiquitous fungal species that contaminates diverse substrates and commodities and produces an array of metabolites toxic to human and animals. The present work has obtained evidence, by liquid chromatography (LC)-ion-trap mass spectrometry, of the ability of P. brevicompactum strains isolated from grapes to produce mycophenolic acid, a potent immunosuppressor. In order to facilitate early diagnosis of this species on commodities for human and animal consumption, a rapid, sensitive and specific polymerase chain reaction (PCR) assay for P. brevicompactum was developed. The specific primers were designed based on the ITS1-5.8S-ITS2ITS (Internal Transcribed Spacers of rRNA genes) multicopy region. This method provides a useful aid to detect the presence of this fungal species in grapes and other commodities in order to prevent the toxins produced entering the food chain.

  4. [Disseminated infection due to Penicillium marneffei related to HIV infection: first observation in Argentina].

    PubMed

    Santiso, Gabriela; Chediak, Viviana; Maiolo, Elena; Mujica, María T; San Juan, Jorge; Arechavala, Alicia; Negroni, Ricardo

    2011-01-01

    The first case observed in Argentina of AIDS-related human penicillosis is herein presented. The patient was a six- teen year-old young man coming from a rural area of southern China. He was admitted at the F. J. Muñiz Hospital of Buenos Aires city with severe pneumonia and adult respiratory distress. Penicillium marneffei was isolated from bronchoalveolar lavage fuid and was microscopically observed in a skin cytodiagnosis. P. marneffei identification was confirmed by rRNA amplification and its phenotypic characteristics. The patient suffered an advanced HIV infection and also presented several AIDS-related diseases due to CMV, nosocomial bacterial infections and Pneumocystis jirovecii which led to a fatal outcome.

  5. Morphogenetic circuitry regulating growth and development in the dimorphic pathogen Penicillium marneffei.

    PubMed

    Boyce, Kylie J; Andrianopoulos, Alex

    2013-02-01

    Penicillium marneffei is an emerging human-pathogenic fungus endemic to Southeast Asia. Like a number of other fungal pathogens, P. marneffei exhibits temperature-dependent dimorphic growth and grows in two distinct cellular morphologies, hyphae at 25°C and yeast cells at 37°C. Hyphae can differentiate to produce the infectious agents, asexual spores (conidia), which are inhaled into the host lung, where they are phagocytosed by pulmonary alveolar macrophages. Within macrophages, conidia germinate into unicellular yeast cells, which divide by fission. This minireview focuses on the current understanding of the genes required for the morphogenetic control of conidial germination, hyphal growth, asexual development, and yeast morphogenesis in P. marneffei.

  6. Bioconversion to chitosan: a two stage process employing chitin deacetylase from Penicillium oxalicum SAEM-51.

    PubMed

    Pareek, Nidhi; Vivekanand, V; Agarwal, Pragati; Saroj, Samta; Singh, Rajesh P

    2013-07-25

    Chitin deacetylase from Penicillium oxalicum SAEM-51 was evaluated for bioconversion of chitin to chitosan in a two stage chemical and enzymatic process. Variations in morphology, crystallinity and thermal properties following chemical treatment were evaluated by scanning electron microscopy, X-ray diffraction, differential scanning calorimetry and thermogravimetric analysis. Degree of deacetylation of the substrates was determined using FT-IR and elemental analysis. The pretreatment of substrate led to the decrease in crystallinity and formation of amorphous chitinous substrates to facilitate enzyme reaction. The treated chitin was further subjected to enzymatic deacetylation employing chitin deacetylase from P. oxalicum SAEM-51 to produce chitosan with considerably higher degree of deacetylation. Maximum deacetylation (79.52%) was achieved using superfine chitin, owing to its porous structure and low crystallinity. Further, derivation of reaction variables, i.e. substrate amount and enzyme dose through full-factorial central composite design led to enhanced degree of deacetylation with formation of 90% deacetylated chitosan.

  7. Bioactive Compounds Produced by Strains of Penicillium and Talaromyces of Marine Origin

    PubMed Central

    Nicoletti, Rosario; Trincone, Antonio

    2016-01-01

    In recent years, the search for novel natural compounds with bioactive properties has received a remarkable boost in view of their possible pharmaceutical exploitation. In this respect the sea is entitled to hold a prominent place, considering the potential of the manifold animals and plants interacting in this ecological context, which becomes even greater when their associated microbes are considered for bioprospecting. This is the case particularly of fungi, which have only recently started to be considered for their fundamental contribution to the biosynthetic potential of other more valued marine organisms. Also in this regard, strains of species which were previously considered typical terrestrial fungi, such as Penicillium and Talaromyces, disclose foreground relevance. This paper offers an overview of data published over the past 25 years concerning the production and biological activities of secondary metabolites of marine strains belonging to these genera, and their relevance as prospective drugs. PMID:26901206

  8. Quantitative quenching evaluation and direct intracellular metabolite analysis in Penicillium chrysogenum.

    PubMed

    Meinert, Sabine; Rapp, Sina; Schmitz, Katja; Noack, Stephan; Kornfeld, Georg; Hardiman, Timo

    2013-07-01

    Sustained progress in metabolic engineering methodologies has stimulated new efforts toward optimizing fungal production strains such as through metabolite analysis of Penicillium chrysogenum industrial-scale processes. Accurate intracellular metabolite quantification requires sampling procedures that rapidly stop metabolism (quenching) and avoid metabolite loss via the cell membrane (leakage). When sampling protocols are validated, the quenching efficiency is generally not quantitatively assessed. For fungal metabolomics, quantitative biomass separation using centrifugation is a further challenge. In this study, P. chrysogenum intracellular metabolites were quantified directly from biomass extracts using automated sampling and fast filtration. A master/slave bioreactor concept was applied to provide industrial production conditions. Metabolic activity during sampling was monitored by 13C tracing. Enzyme activities were efficiently stopped and metabolite leakage was absent. This work provides a reliable method for P. chrysogenum metabolomics and will be an essential base for metabolic engineering of industrial processes.

  9. Morphogenesis and Production of Enzymes by Penicillium echinulatum in Response to Different Carbon Sources

    PubMed Central

    Schneider, Willian Daniel Hahn; dos Reis, Laísa; Dillon, Aldo José Pinheiro

    2014-01-01

    The effect of different carbon sources on morphology and cellulase and xylanase production of Penicillium echinulatum was evaluated in this work. Among the six carbon sources studied, cellulose and sugar cane bagasse were the most suitable for the production of filter paper activity, endoglucanases, xylanases, and β-glucosidases. However, sucrose and glucose showed β-glucosidase activities similar to those obtained with the insoluble sources. The polyacrylamide gels proved the enzymatic activity, since different standards bands were detected in the media mentioned above. Regarding morphology, it was observed that the mycelium in a dispersed form provided the greatest enzymatic activity, possibly due to greater interaction between the substrate and hyphae. These data are important in understanding the physiology of fungi and could contribute to obtaining enzyme with potential application in the technology of second generation ethanol. PMID:24877074

  10. Autolysis and ageing of Penicillium chrysogenum under carbon starvation: respiration and glucose oxidase production.

    PubMed

    Sámi, L; Karaffa, L; Emri, T; Pócsi, I

    2003-01-01

    During the exponential growth phase of Penicillium chrysogenum NCAIM 00237 the effective conversion of glucose and O2 to gluconate and H2O2 by glucose oxidase (GOX) was the most likely source of intracellular ROS measured. In glucose-supplemented autolysing cultures, the increased of intracellular ROS concentration was attributed to respiration in the absence of any significant GOX activity. The induction of GOX and catalase by glucose and H2O2 was clearly age-dependent in P. chrysogenum. In ageing cryptic growth phase cultures, superoxide dismutase and cyanide-resistant respiration were the major elements of antioxidative defence but these activities were insufficient to prevent the progressive accumulation of ROS and the concomitant decrease in cell vitality.

  11. Biosynthetic concepts for the production of β-lactam antibiotics in Penicillium chrysogenum.

    PubMed

    Weber, Stefan S; Bovenberg, Roel A L; Driessen, Arnold J M

    2012-02-01

    Industrial production of β-lactam antibiotics by the filamentous fungus Penicillium chrysogenum is based on successive classical strain improvement cycles. This review summarizes our current knowledge on the results of this classical strain improvement process, and discusses avenues to improve β-lactam biosynthesis and to exploit P. chrysogenum as an industrial host for the production of other antibiotics and peptide products. Genomic and transcriptional analysis of strain lineages has led to the identification of several important alterations in high-yielding strains, including the amplification of the penicillin biosynthetic gene cluster, elevated transcription of genes involved in biosynthesis of penicillin and amino acid precursors, and genes encoding microbody proliferation factors. In recent years, successful metabolic engineering and synthetic biology approaches have resulted in the redirection of the penicillin pathway towards the production of cephalosporins. This sets a new direction in industrial antibiotics productions towards more sustainable methods for the fermentative production of unnatural antibiotics and related compounds.

  12. The ABC transporter ABC40 encodes a phenylacetic acid export system in Penicillium chrysogenum.

    PubMed

    Weber, Stefan S; Kovalchuk, Andriy; Bovenberg, Roel A L; Driessen, Arnold J M

    2012-11-01

    The filamentous fungus Penicillium chrysogenum is used for the industrial production of β-lactam antibiotics. The pathway for β-lactam biosynthesis has been resolved and involves the enzyme phenylacetic acid CoA ligase that is responsible for the CoA activation of the side chain precursor phenylacetic acid (PAA) that is used for the biosynthesis of penicillin G. To identify ABC transporters related to β-lactam biosynthesis, we analyzed the expression of all 48 ABC transporters present in the genome of P. chryso-genum when grown in the presence and absence of PAA. ABC40 is significantly upregulated when cells are grown or exposed to high levels of PAA. Although deletion of this transporter did not affect β-lactam biosynthesis, it resulted in a significant increase in sensitivity to PAA and other weak acids. It is concluded that ABC40 is involved in weak acid detoxification in P. chrysogenum including resistance to phenylacetic acid.

  13. New compound with DNA Topo I inhibitory activity purified from Penicillium oxalicum HSY05.

    PubMed

    Liu, Bing; Wang, Hai-Feng; Zhang, Li-Hua; Liu, Fang; He, Feng-Jun; Bai, Jiao; Hua, Hui-Ming; Chen, Gang; Pei, Yue-Hu

    2015-01-01

    Strain HSY05 was isolated from sea sediment collected from the South China Sea and was later identified as Penicillium oxalicum by 16S rDNA sequence analysis. Various chromatographic processes led to the isolation and purification of two metabolites from the fermentation culture of HSY05, including one new compound, 2,2',4,4'-tetrahyoxy-8'-methyl-6-methoxy-acyl-ethyl-diphenylmethanone (1), and a known compound secalonic acid D (SAD, 2), as characterised by UV, IR, 1D, 2D-NMR and MS data. The inhibitory activities against topoisomerase I of these two compounds were evaluated. The result showed that in addition to the known topo I inhibitor SAD (2), compound 1 also exhibited a moderate inhibitory effect.

  14. Degradation of 2,4-dichlorophenoxyacetic acid by a halotolerant strain of Penicillium chrysogenum: antibiotic production.

    PubMed

    Ferreira-Guedes, Sumaya; Mendes, Benilde; Leitão, Ana Lúcia

    2012-01-01

    The extensive use of pesticides in agriculture has prompted intensive research on chemical and biological methods in order to protect contamination of water and soil resources. In this paper the degradation of the pesticide 2,4-dichlorophenoxyacetic acid by a Penicillium chrysogenum strain previously isolated from a salt mine was studied in batch cultures. Co-degradation of 2,4-dichlorophenoxyacetic acid with additives such as sugar and intermediates of pesticide metabolism was also investigated. Penicillium chrysogenum in solid medium was able to grow at concentrations up to 1000 mg/L of 2,4-dichlorophenoxyacetic acid (2,4-D) with sucrose. Meanwhile, supplementation of the solid medium with glucose and lactose led to fungal growth at concentrations up to 500 mg/L of herbicide. Batch cultures of 2,4-D at 100 mg/L were developed under aerobic conditions with the addition of glucose, lactose and sucrose, showing sucrose as the best additional carbon source. The 2,4-D removal was quantified by liquid chromatography. The fungus was able to use 2,4-D as the sole carbon and energy source under 0%, 2% and 5.9% NaCl. The greatest 2,4-D degradation efficiency was found using alpha-ketoglutarate and ascorbic acid as co-substrates under 2% NaCl at pH 7. Penicillin production was evaluated in submerged cultures by bioassay, and higher amounts of beta-lactam antibiotic were produced when the herbicide was alone. Taking into account the ability of P. chrysogenum CLONA2 to degrade aromatic compounds, this strain could be an interesting tool for 2,4-D herbicide remediation in saline environments.

  15. Lag time for germination of Penicillium chrysogenum conidia is induced by temperature shifts.

    PubMed

    Kalai, Safaa; Bensoussan, Maurice; Dantigny, Philippe

    2014-09-01

    In the environment, fungal conidia are subject to transient conditions. In particular, temperature is varying according to day/night periods. All predictive models for germination assume that fungal spores can adapt instantaneously to changes of temperature. The only study that supports this assumption (Gougouli and Koutsoumanis, 2012, Modelling germination of fungal spores at constant and fluctuating temperature conditions. International Journal of Food Microbiology, 152: 153-161) was carried out on Penicillium expansum and Aspergillus niger conidia that, in most cases, already produced germ tubes. In contrast, the present study focuses on temperature shifts applied during the first stages of germination (i.e., before the apparition of the germ tubes). Firstly, germination times were determined in steady state conditions at 10, 15, 20 and 25 °C. Secondly, temperature shifts (e.g., up-shifts and down-shifts) were applied at 1/4, 1/2, and 3/4 of germination times, with 5, 10 and 15 °C magnitudes. Experiments were carried out in triplicate on Penicillium chrysogenum conidia on Potato Dextrose Agar medium according to a full factorial design. Statistical analysis of the results clearly demonstrated that the assumption of instantaneous adaptation of the conidia should be rejected. Temperature shifts during germination led to an induced lag time or an extended germination time as compared to the experiments conducted ay steady state. The induced lag time was maximized when the amplitude of the shift was equal to 10 °C. Interaction between the instant and the direction of the shift was highlighted. A negative lag time was observed for a 15 °C down-shift applied at 1/4 of the germination time. This result suggested that at optimal temperature the rate of germination decreased with time, and that the variation of this rate with time depended on temperature.

  16. Cloning, recombinant expression and characterization of a new phytase from Penicillium chrysogenum.

    PubMed

    Ribeiro Corrêa, Thamy Lívia; de Queiroz, Marisa Vieira; de Araújo, Elza Fernandes

    2015-01-01

    The phy gene, which encodes a phytase in Penicillium chrysogenum CCT 1273, was cloned into the vector pAN-52-1-phy and the resulting plasmid was used for the cotransformation of Penicillium griseoroseum PG63 protoplasts. Among the 91 transformants obtained, 23 were cotransformants. From there, the phytase activity of these 23 transformants was evaluated and P. griseoroseum T73 showed the highest. The recombinant strain P. griseoroseum T73 contained the phy gene integrated in at least three sites of the genome and showed a 5.1-fold increase in phytase activity in comparison to the host strain (from 0.56 ± 0.2 to 2.86 ± 0.4 U μg protein(-1)). The deduced PHY protein has 483 amino acids; an isoelectric point (pI) higher than that reported for phytases from filamentous fungi (7.6); higher activity at pH 2.0 (73%), pH 5.0 (100%) and 50 °C; and is stable at pH values 3.0-8.0 and temperatures 70-80 °C. PHY produced by the recombinant strain P. griseoroseum T73 was stable after four weeks of storage at -20, 8 and 25 °C and was effective in releasing Pi, especially from soybeans. The data presented here show that P. griseoroseum is a successful host for expression of heterologous protein and suggest the potential use of PHY in the animal nutrition industry.

  17. Synergistic antifungal activity of sodium hypochlorite, hydrogen peroxide, and cupric sulfate against Penicillium digitatum.

    PubMed

    Cerioni, Luciana; Rapisarda, Viviana Andrea; Hilal, Mirna; Prado, Fernando Eduardo; Rodríguez-Montelongo, Luisa

    2009-08-01

    Oxidizing compounds such as sodium hypochlorite (NaCIO) and hydrogen peroxide (H2O2) are widely used in food sanitization because of their antimicrobial effects. We applied these compounds and metals to analyze their antifungal activity against Penicillium digitatum, the causal agent of citrus green mold. The MICs were 300 ppm for NaClO and 300 mM for H2O2 when these compounds were individually applied for 2 min to conidia suspensions. To minimize the concentration of these compounds, we developed and standardized a sequential treatment for conidia that resulted in loss of viability on growth plates and loss of infectivity on lemons. The in vitro treatment consists of preincubation with 10 ppm of NaClO followed by incubation with 100 mM H2O2 and 6 mM CuSO4 (cupric sulfate). The combination of NaClO and H2O2 in the presence of CuSO4 produces a synergistic effect (fractional inhibitory concentration index of 0.36). The sequential treatment applied in situ on lemon peel 24 h after the fruit was inoculated with conidia produced a significant delay in the fungal infection. The in vitro treatment was effective on both imazalil-sensitive and imazalil-resistant strains of P. digitatum and Geotrichum candidum, the causal agent of citrus sour rot. However, this treatment inhibited 90% of mycelial growth for Penicillium italicum (citrus blue mold). These results indicate that sequential treatment may be useful for postharvest control of citrus fruit diseases.

  18. Purification and biochemical characterization of a novel alkaline protease produced by Penicillium nalgiovense.

    PubMed

    Papagianni, M; Sergelidis, D

    2014-04-01

    Penicillium nalgiovense PNA9 produces an extracellular protease during fermentation with characteristics of growth-associated product. Enzyme purification involved ammonium sulfate precipitation, dialysis, and ultrafiltration, resulting in 12.1-fold increase of specific activity (19.5 U/mg). The protein was isolated through a series of BN-PAGE and native PAGE runs. ESI-MS analysis confirmed the molecular mass of 45.2 kDa. N-Terminal sequencing (MGFLKLLKGSLATLAVVNAGKLLTANDGDE) revealed 93 % similarity to a Penicillium chrysogenum protease, identified as major allergen. The protease exhibits simple Michaelis-Menten kinetics and K m (1.152 mg/ml), V max (0.827 mg/ml/min), and k cat (3.2 × 10(2)) (1/s) values against azocasein show that it possesses high substrate affinity and catalytic efficiency. The protease is active within 10-45 °C, pH 4.0-10.0, and 0-3 M NaCl, while maximum activity was observed at 35 °C, pH 8.0, and 0.25 M NaCl. It is active against the muscle proteins actin and myosin and inactive against myoglobin. It is highly stable in the presence of non-ionic surfactants, hydrogen peroxide, BTNB, and EDTA. Activity was inhibited by SDS, Mn(2+) and Zn(2+), and by the serine protease inhibitor PMSF, indicating the serine protease nature of the enzyme. These properties make the novel protease a suitable candidate enzyme in meat ripening and other biotechnological applications.

  19. Effects of different culture conditions on biological potential and metabolites production in three Penicillium isolates.

    PubMed

    Reis, Filipa S; Ćirić, Ana; Stojković, Dejan; Barros, Lillian; Ljaljević-Grbić, Milica; Soković, Marina; Ferreira, Isabel C F R

    2015-02-01

    The genus Penicillium is well known for its importance in drug and food production. Certain species are produced on an industrial scale for the production of antibiotics (e.g. penicillin) or for insertion in food (e.g. cheese). In the present work, three Penicillium species, part of the natural mycobiota growing on various food products were selected - P. ochrochloron, P. funiculosum and P. verrucosum var. cyclopium. The objective of our study was to value these species from the point of view of production of bioactive metabolites. The species were obtained after inoculation and growth in Czapek and Malt media. Both mycelia and culture media were analyzed to monitor the production of different metabolites by each fungus and their release to the culture medium. The concentrations of sugars, organic acids, phenolic acids and tocopherols were determined. Antioxidant activity of the phenolic extracts was evaluated, as also the antimicrobial activity of phenolic acids, organic acids and tocopherols extracts. Rhamnose, xylose, fructose and trehalose were found in all the mycelia and culture media; the prevailing organic acids were oxalic and fumaric acids, and protocatechuic and p-hydroxybenzoic acids were the most common phenolic acids; γ-tocopherol was the most abundant vitamin E isoform. Generally, the phenolic extracts corresponding to the mycelia samples revealed higher antioxidant activity. Concerning the antimicrobial activity there were some fluctuations, however all the studied species revealed activity against the tested strains. Therefore, the in-vitro bioprocesses can be an alternative for the production of bioactive metabolites that can be used by pharmaceutical industry.

  20. In vivo application of a small molecular weight antifungal protein of Penicillium chrysogenum (PAF)

    SciTech Connect

    Palicz, Zoltán; Jenes, Ágnes; Gáll, Tamás; Miszti-Blasius, Kornél; Kollár, Sándor; Kovács, Ilona; Emri, Miklós; Márián, Teréz; Leiter, Éva; Pócsi, István; Csősz, Éva; Kalló, Gergő; Hegedűs, Csaba; Virág, László; Csernoch, László; Szentesi, Péter

    2013-05-15

    The antifungal protein of Penicillium chrysogenum (PAF) inhibits the growth of important pathogenic filamentous fungi, including members of the Aspergillus family and some dermatophytes. Furthermore, PAF was proven to have no toxic effects on mammalian cells in vitro. To prove that PAF could be safely used in therapy, experiments were carried out to investigate its in vivo effects. Adult mice were inoculated with PAF intranasally in different concentrations, up to 2700 μg·kg{sup −1} daily, for 2 weeks. Even at the highest concentration – a concentration highly toxic in vitro for all affected molds – used, animals neither died due to the treatment nor were any side effects observed. Histological examinations did not find pathological reactions in the liver, in the kidney, and in the lungs. Mass spectrometry confirmed that a measurable amount of PAF was accumulated in the lungs after the treatment. Lung tissue extracts from PAF treated mice exerted significant antifungal activity. Small-animal positron emission tomography revealed that neither the application of physiological saline nor that of PAF induced any inflammation while the positive control lipopolysaccharide did. The effect of the drug on the skin was examined in an irritative dermatitis model where the change in the thickness of the ears following PAF application was found to be the same as in control and significantly less than when treated with phorbol-12-myristate-13-acetate used as positive control. Since no toxic effects of PAF were found in intranasal application, our result is the first step for introducing PAF as potential antifungal drug in therapy. - Highlights: • PAF, the antifungal protein of Penicillium chrysogenum, was not toxic in mice. • Its intranasal application didn't induce pathological reactions in the lung. • PAF retained its antifungal activity in lung extracts. • Its application on the skin did not cause inflammation.

  1. Genome sequence of the necrotrophic fungus Penicillium digitatum, the main postharvest pathogen of citrus

    PubMed Central

    2012-01-01

    Background Penicillium digitatum is a fungal necrotroph causing a common citrus postharvest disease known as green mold. In order to gain insight into the genetic bases of its virulence mechanisms and its high degree of host-specificity, the genomes of two P. digitatum strains that differ in their antifungal resistance traits have been sequenced and compared with those of 28 other Pezizomycotina. Results The two sequenced genomes are highly similar, but important differences between them include the presence of a unique gene cluster in the resistant strain, and mutations previously shown to confer fungicide resistance. The two strains, which were isolated in Spain, and another isolated in China have identical mitochondrial genome sequences suggesting a recent worldwide expansion of the species. Comparison with the closely-related but non-phytopathogenic P. chrysogenum reveals a much smaller gene content in P. digitatum, consistent with a more specialized lifestyle. We show that large regions of the P. chrysogenum genome, including entire supercontigs, are absent from P. digitatum, and that this is the result of large gene family expansions rather than acquisition through horizontal gene transfer. Our analysis of the P. digitatum genome is indicative of heterothallic sexual reproduction and reveals the molecular basis for the inability of this species to assimilate nitrate or produce the metabolites patulin and penicillin. Finally, we identify the predicted secretome, which provides a first approximation to the protein repertoire used during invasive growth. Conclusions The complete genome of P. digitatum, the first of a phytopathogenic Penicillium species, is a valuable tool for understanding the virulence mechanisms and host-specificity of this economically important pest. PMID:23171342

  2. Screening and optimization of some inorganic salts for the production of ergot alkaloids from Penicillium species using surface culture fermentation process.

    PubMed

    Shahid, Memuna Ghafoor; Nadeem, Muhammad; Baig, Shahjehan; Cheema, Tanzeem Akbar; Atta, Saira; Ghafoor, Gul Zareen

    2016-03-01

    The present study deals with the production of ergot alkaloids from Penicillium commune and Penicillium citrinum, using surface culture fermentation process. Impact of various inorganic salts was tested on the production of ergot alkaloids during the optimization studies of fermentation medium such as impact of various concentration levels of succinic acid, ammonium chloride, MgSO4, FeSO4, ZnSO4, pH and the effect of various incubation time periods was also determined on the production of ergot alkaloids from Penicillium commune and Penicillium citrinum. Highest yield of ergot alkaloids was obtained when Penicillium commune and Penicillium citrinum that were grown on optimum levels of ingredients such as 2 g succinic acid, 1.5 and 2 g NH4Cl, 1.5 g MgSO4, 1 g FeSO4, 1 and 1.5 g ZnSO4 after 21 days of incubation time period using pH 5 at 25(°)C incubation temperature in the fermentation medium. Ergot alkaloids were determined using Spectrophotometry and Thin Layer Chromatography (TLC) techniques.

  3. Characterization of a 52 kDa exoantigen of Penicillium chrysogenum and monoclonal antibodies suitable for its detection.

    PubMed

    Luo, Wen; Wilson, Aaron M; Miller, J David

    2010-01-01

    The indoor clade of Penicillium chrysogenum, the so-called Fleming clade, is the most common species of Penicillium on moldy building materials. In a previous study, we identified a 52 kDa human antigen characteristic of the indoor clade of P. chrysogenum not present in a taxonomically diverse selection of fungi. Further investigations revealed that it is a modestly glycosylated mature protein with a pI 5.3. The protein is apparently identical to a glucoamylase previously reported from an aluminum-tolerant P. chrysogenum mutant. Based on sequence similarity, molecular weight, and pI, it is distinct from a number of other glucoamylases from domesticated strains of Aspergillus oryzae and A. niger used to produce industrial enzymes. Surprisingly, it had not been reported as an allergen. The monoclonal antibodies developed have the potential for use in assays of P. chrysogenum antigens in spores and spore/mycelial fragments in dust.

  4. The effect of acetylated xylan and sugar beet pulp on the expression and secretion of enzymes by Penicillium purpurogenum.

    PubMed

    Navarrete, Mario; Callegari, Eduardo; Eyzaguirre, Jaime

    2012-01-01

    Sugar beet pulp is a natural carbon source composed mainly of pectin and cellulose, which is utilized and degraded by the ascomycete Penicillium purpurogenum. The fungus also grows on and degrades acetylated xylan which lacks cellulose and pectin. Both carbon sources have been used in our laboratory to grow the fungus and to purify different enzymes secreted to the medium. The enzymes involved in the complex process of degradation of these carbon sources by the fungus have been explored previously under non-denaturing conditions; multienzyme complexes were separated and some subunits identified by Western blots and mass spectrometry. In this work, proteomic profiles show that the secretome is composed of numerous proteins varying in pI and molecular weight. Some enzymes are common to both growth conditions, while others are specific for each carbon source. The results show that the carbon sources utilized exert strong regulatory control over the proteins secreted. This is the first secretome study from a lignocellulolytic Penicillium.

  5. Influence of temperature, pH and water activity on "in vitro" inhibition of Penicillium glabrum (Wehmer) Westling by yeasts.

    PubMed

    Sinigaglia, M; Corbo, M R; Ciccarone, C

    1998-08-01

    Four different yeast species (Metschnikowia pulcherrima, Saccharomycopsis vini, Kluyveromyces marxianus, Cryptococcus albidus), isolated from surface of grapes, were evaluated for biocontrol potential against Penicillium glabrum. In order to investigate the influence of temperature, pH, water activity and yeast cell concentration on Penicillium glabrum inhibition, the individual effects and the interaction of these factors were analyzed by means of a Central Composite Design (CCD). All yeast species tested showed antagonistic effects which were more pronounced at high cell concentrations. The other variables affected the antagonistic effect differentially depending on the yeast species. Results of the experimental design showed that the selective success of a competitive microflora is under environmental control; moreover, when microbial cells are subjected to multiple factors, the effects and the reciprocal interactions of the individual variables cannot be independently evaluated.

  6. A retrospective review on successful management of Penicillium marneffei infections in patients with advanced HIV in Hospital Sungai Buloh.

    PubMed

    Nor-Hayati, S; Sahlawati, M; Suresh-Kumar, C; Lee, K C Christopher

    2012-02-01

    Penicillium marneffei is a dimorphic fungus which commonly causes a life threatening systemic fungal infection in an immunocompromised host. It has been recognized as an AIDS defining illness in Malaysia since the beginning of the HIV pandemic. The presence of various non specific clinical presentations, especially the characteristic umbilicated papular rashes with central necrosis which lead to significant ill health in immunocompromised patients should alarm clinicians to the possibility of Penicillium marneffei infection and prompt investigations accordingly. Simple investigations like blood culture and fungal staining of the skin scrapping can confirm the diagnosis in the majority of cases. Early treatment with appropriate systemic antifungal for a definite duration will significantly decrease the mortality rate from penicilliosis.

  7. Microbial transformations of testosterone to 5alpha-dihydrotestosterone by two species of Penicillium: P. chrysogenum and P. crustosum.

    PubMed

    Cabeza, M S; Gutiérrez, E B; García, G A; Avalos, A H; Hernández, M A

    1999-06-01

    Two species of Penicillium--P. chrysogenum and P. crustosum--were cultured in presence of [3H]testosterone as a substrate. Both species were shown to reduce the 4,5-double bond in testosterone to give dihydrotestosterone (DHT). The steroids produced were 5alpha-dihydrotestosterone, DHT, 3alpha-hydroxy-5beta-androstan-17-one, 3alpha-hydroy-5alpha-androstan-17-one, 4-androstene-3,17-dione, and 5alpha-androstane-3,17-dione. These products implicate the presence of the 5alpha-reductase, with maximal activity at pH 6 and 8, in both species of Penicillium. The presence of DHT in the growth medium and not in the mycelium suggests that DHT is excreted into the medium.

  8. Cytotoxic 1,3-Thiazole and 1,2,4-Thiadiazole Alkaloids from Penicillium oxalicum: Structural Elucidation and Total Synthesis.

    PubMed

    Yang, Zheng; Huang, Nianyu; Xu, Bang; Huang, Wenfeng; Xie, Tianpeng; Cheng, Fan; Zou, Kun

    2016-02-26

    Two new thiazole and thiadiazole alkaloids, penicilliumthiamine A and B (2 and 3), were isolated from the culture broth of Penicillium oxalicum, a fungus found in Acrida cinerea. Their structures were elucidated mainly by spectroscopic analysis, total synthesis and X-ray crystallographic analysis. Biological evaluations indicated that compound 1, 3a and 3 exhibit potent cytotoxicity against different cancer cell lines through inhibiting the phosphorylation of AKT/PKB (Ser 473), one of important cancer drugs target.

  9. Genome Sequencing and Comparative Genomics Analysis Revealed Pathogenic Potential in Penicillium capsulatum as a Novel Fungal Pathogen Belonging to Eurotiales

    PubMed Central

    Yang, Ying; Chen, Min; Li, Zongwei; Al-Hatmi, Abdullah M. S.; de Hoog, Sybren; Pan, Weihua; Ye, Qiang; Bo, Xiaochen; Li, Zhen; Wang, Shengqi; Wang, Junzhi; Chen, Huipeng; Liao, Wanqing

    2016-01-01

    Penicillium capsulatum is a rare Penicillium species used in paper manufacturing, but recently it has been reported to cause invasive infection. To research the pathogenicity of the clinical Penicillium strain, we sequenced the genomes and transcriptomes of the clinical and environmental strains of P. capsulatum. Comparative analyses of these two P. capsulatum strains and close related strains belonging to Eurotiales were performed. The assembled genome sizes of P. capsulatum are approximately 34.4 Mbp in length and encode 11,080 predicted genes. The different isolates of P. capsulatum are highly similar, with the exception of several unique genes, INDELs or SNPs in the genes coding for glycosyl hydrolases, amino acid transporters and circumsporozoite protein. A phylogenomic analysis was performed based on the whole genome data of 38 strains belonging to Eurotiales. By comparing the whole genome sequences and the virulence-related genes from 20 important related species, including fungal pathogens and non-human pathogens belonging to Eurotiales, we found meaningful pathogenicity characteristics between P. capsulatum and its closely related species. Our research indicated that P. capsulatum may be a neglected opportunistic pathogen. This study is beneficial for mycologists, geneticists and epidemiologists to achieve a deeper understanding of the genetic basis of the role of P. capsulatum as a newly reported fungal pathogen. PMID:27761131

  10. Origin of (-)-geosmin on grapes: on the complementary action of two fungi, botrytis cinerea and penicillium expansum.

    PubMed

    La Guerche, Stéphane; Chamont, Sophie; Blancard, Dominique; Dubourdieu, Denis; Darriet, Philippe

    2005-08-01

    One of the consequences of rot on grapes is the development of volatile compounds giving fungal, mouldy or earthy odours. Among these compounds, (-)-geosmin (trans-1,10-dimethyl-trans-9-decalol), a powerful aromatic compound with an earthy smell is a persistent defect in grape juice and wines made with at least partially rotten grapes. A microbiota analysis of rotten grapes containing (-)-geosmin was carried out on sites from four French regions from 1999 to 2002, to clarify the involvement in geosmin appearance of Streptomyces spp. and Penicillium spp., two types of microorganisms present on grape, that are known for their ability to produce geosmin. In earthy grapes, Botrytis cinerea was largely present. Different species of Streptomyces were also isolated, but their pH sensitivity was an extremely limiting parameter for their development on grape juice, grapes or stem, and consequently for their potentiality to generate geosmin in the vineyard. Penicillium expansum, producing geosmin on a model medium, was omnipresent. Penicillium carneum, which is also a geosmin producer, was represented by a single colony during the 4 years of this study. P. expansum alone was able to produce geosmin on a model medium but not on grapes. However, after 7 days' pre-culture of some B. cinerea strains on grape juice, this juice became favourable to geosmin production by P. expansum. We demonstrated the necessary and complementary action of B. cinerea and P. expansum in geosmin production in grape juice and in crushed grape berries.

  11. The use of AFLP to relate cheese-contaminating Penicillium strains to specific points in the production plants.

    PubMed

    Kure, C F; Skaar, I; Holst-Jensen, A; Abeln, E C A

    2003-06-15

    Amplified fragment length polymorphism (AFLP) analysis was performed on isolates of Penicillium commune and Penicillium palitans originating from cheese and indoor environment in four cheese factories. The AFLP method was found to be a useful tool for identification of P. commune and P. palitans on, as well as below, species level. However, AFLP in combination with M13 fingerprinting described in a previous paper provided better resolution at the intraspecific level than either of the methods alone. Specific P. commune and P. palitans strains were found in the same factories over a period of more than a year and showed that the cheese factories have contaminating strains that are well established. The majority of the P. commune and P. palitans strains were found only within a single factory, but several were found in different cheese factories. The combined fingerprinting data could relate strains isolated from cheese to specific points in the production plants. Several of cheese-contaminating Penicillium strains could be related to air in the wrapping room, which must be considered to be a critical point for contamination of cheese.

  12. FORMATION OF TRYPSIN FROM TRYPSINOGEN BY AN ENZYME PRODUCED BY A MOLD OF THE GENUS PENICILLIUM

    PubMed Central

    Kunitz, M.

    1938-01-01

    1. A powerful kinase which changes trypsinogen to trypsin was found to be present in the synthetic liquid culture medium of a mold of the genus Penicillium. 2. The concentration of kinase in the medium is increased gradually during the growth of the mold organism and continues to increase for some time even after the mold has ceased growing. 3. Mold kinase transforms trypsinogen to trypsin only in an acid medium. It differs thus from enterokinase and trypsin which activate trypsinogen best in a slightly alkaline medium. 4. The action of the mold kinase in the process of transformation of trypsinogen is that of a typical enzyme. The process follows the course of a catalytic unimolecular reaction, the rate of formation of a definite amount of trypsin being proportional to the concentration of kinase added. The ultimate amount of trypsin formed, however, is independent of the concentration of kinase used. 5. The formation of trypsin from trypsinogen by mold kinase is not accompanied by any measurable loss of protein. 6. The temperature coefficient of formation of trypsin from trypsinogen by mold kinase varies from Q5–15 = 1.70 to Q25–30 = 1.25 with a corresponding variation in the value of µ from 8100 to 4250. 7. Trypsin formed from trypsinogen by means of mold kinase is identical in crystalline form with the crystalline trypsin obtained by spontaneous autocatalytic activation of trypsinogen at pH 8.0. The two products have within the experimental error the same solubility and specific activity. A solution saturated with the crystals of either one of the trypsin preparations does not show any increase in protein concentration or activity when crystals of the other trypsin preparation are added. 8. The Penicillium mold kinase has a slight activating effect on chymo-trypsinogen the rate being only 1–2 per cent of that of trypsinogen. The activation, as in the case of trypsinogen, takes place only in an acid medium. 9. Mold kinase is rapidly destroyed when brought

  13. A natural short pathway synthesizes roquefortine C but not meleagrin in three different Penicillium roqueforti strains.

    PubMed

    Kosalková, K; Domínguez-Santos, R; Coton, M; Coton, E; García-Estrada, C; Liras, P; Martín, J F

    2015-09-01

    The production of mycotoxins and other secondary metabolites in Penicillium roqueforti is of great interest because of its long history of use in blue-veined cheese manufacture. In this article, we report the cloning and characterization of the roquefortine gene cluster in three different P. roqueforti strains isolated from blue cheese in the USA (the type strain), France, and the UK (Cheshire cheese). All three strains showed an identical roquefortine gene cluster organization and almost identical (98-99%) gene nucleotide sequences in the entire 16.6-kb cluster region. When compared with the Penicillium chrysogenum roquefortine/meleagrin seven-gene cluster, the P. roqueforti roquefortine cluster contains only four genes (rds, rdh, rpt, and gmt) encoding the roquefortine dipeptide synthetase, roquefortine D dehydrogenase, roquefortine prenyltransferase, and a methyltransferase, respectively. Silencing of the rds or rpt genes by the RNAi strategy reduced roquefortine C production by 50% confirming the involvement of these two key genes in roquefortine biosynthesis. An additional putative gene, orthologous of the MFS transporter roqT, is rearranged in all three strains as a pseudogene. The same four genes and a complete (not rearranged) roqT, encoding a MFS transporter containing 12 TMS domains, occur in the seven-gene cluster in P. chrysogenum although organized differently. Interestingly, the two "late" genes of the P. chrysogenum roquefortine/meleagrin gene cluster that convert roquefortine C to glandicoline B and meleagrin are absent in the P. roqueforti four-gene cluster. No meleagrin production was detected in P. roqueforti cultures grown in YES medium, while P. chrysogenum produces meleagrin in these conditions. No orthologous genes of the two missing meleagrin synthesizing genes were found elsewhere in the recently released P. roqueforti genome. Our data suggest that during evolution, the seven-gene cluster present in P. chrysogenum, and probably also in

  14. Functional characterization of a Penicillium chrysogenum mutanase gene induced upon co-cultivation with Bacillus subtilis

    PubMed Central

    2014-01-01

    Background Microbial gene expression is strongly influenced by environmental growth conditions. Comparison of gene expression under different conditions is frequently used for functional analysis and to unravel regulatory networks, however, gene expression responses to co-cultivation with other microorganisms, a common occurrence in nature, is rarely studied under laboratory conditions. To explore cellular responses of the antibiotic-producing fungus Penicillium chrysogenum to prokaryotes, the present study investigates its transcriptional responses during co-cultivation with Bacillus subtilis. Results Steady-state glucose-limited chemostats of P. chrysogenum grown under penillicin-non-producing conditions were inoculated with B. subtilis. Physiological and transcriptional responses of P. chrysogenum in the resulting mixed culture were monitored over 72 h. Under these conditions, B. subtilis outcompeted P. chrysogenum, as reflected by a three-fold increase of the B. subtilis population size and a two-fold reduction of the P. chrysogenum biomass concentration. Genes involved in the penicillin pathway and in synthesis of the penicillin precursors and side-chain were unresponsive to the presence of B. subtilis. Moreover, Penicillium polyketide synthase and nonribosomal peptide synthase genes were either not expressed or down-regulated. Among the highly responsive genes, two putative α-1,3 endoglucanase (mutanase) genes viz Pc12g07500 and Pc12g13330 were upregulated by more than 15-fold and 8-fold, respectively. Measurement of enzyme activity in the supernatant of mixed culture confirmed that the co-cultivation with B. subtilis induced mutanase production. Mutanase activity was neither observed in pure cultures of P. chrysogenum or B. subtilis, nor during exposure of P. chrysogenum to B. subtilis culture supernatants or heat-inactivated B. subtilis cells. However, mutanase production was observed in cultures of P. chrysogenum exposed to filter-sterilized supernatants

  15. Cross-reactivity among antigens of different air-borne fungi detected by ELISA using five monoclonal antibodies against Penicillium notatum.

    PubMed

    Shen, H D; Lin, W L; Chen, R J; Han, S H

    1990-10-01

    Cross-reactivity among antigens of 12 genera of air-borne fungi, 13 species of Penicillium, and 5 species of Aspergillus was studied by ELISA using five monoclonal antibodies (MoAbs) against Penicillium notatum. Epitopes recognized by all the five MoAbs were susceptible to treatment of mild periodate oxidation and may therefore be associated with carbohydrates. Furthermore, our results showed that there is cross-reactivity among antigens of Penicillium, Aspergillus, and Eurotium species. By using these MoAbs, cross reactivity was not detected between antigens of Penicillium notatum and antigens of Fusarium solani, Alternaria porri, Cladosporium cladosporoides, Curvularia species, Nigrospora species, Aureobasidium pullulans, Wallemia species, Rhizopus arrhizus, and Candida albicans. Cross-reactivity among antigens of 11 species of Penicillium and 5 species of Aspergillus could be detected by ELISA using one of the five MoAbs (MoAb P15). The fact that there may be cross-reactivity among antigens of closely related fungi species should be considered in the diagnosis and treatment of mold allergic diseases.

  16. Effects of the protective, curative, and eradicative applications of chitosan against Penicillium expansum in apples.

    PubMed

    Darolt, Josiane Cecília; Rocha Neto, Argus Cezar da; Di Piero, Robson Marcelo

    Apple is one of the most important temperate fruit to Brazil economy, and the use of synthetic chemicals has been the main method for reducing postharvest diseases, such as the blue mold, caused by Penicillium expansum. This work intends to evaluate the practical utilization of chitosan for blue mold control. For this purpose, fruits were treated in a preventive and curative way, immersing the fruits in chitosan solution (5 or 10mgmL(-1)), or adding a single drop of this solution (10mgmL(-1)) directly into the injuries. The eradicative effect of the polysaccharide was also evaluated in vitro and in vivo. Chitosan did not show a curative effect against the blue mold, and its eradicative effect was only evidenced on the higher concentration (10mgmL(-1)). On the other hand, preventively, without the addition of adjuvants, chitosan reduced blue mold incidence in fruits by 24% and 93%, through the immersion or the single drop methods, respectively. Thus, it was found that, for long scale utilization, some improvements in the physico-chemical properties of the chitosan are needed, since it was only capable to prevent the infection by P. expansum when directly added on the fruit injury.

  17. Comparative metabolomic study of Penicillium chrysogenum during pilot and industrial penicillin fermentations.

    PubMed

    Ding, Ming-Zhu; Lu, Hua; Cheng, Jing-Sheng; Chen, Yao; Jiang, Jing; Qiao, Bin; Li, Bing-Zhi; Yuan, Ying-Jin

    2012-11-01

    Comparative metabolomics was carried out to investigate the metabolic differences of Penicillium chrysogenum in the pilot and industrial fermentations that resulted from the scale-up. By principal component analysis, the early stages of two fermentation processes were clearly distinguished, whereas the middle and final stages were clustered together. It indicated that the different metabolisms of cells in the pilot and industrial fermentations mainly existed during the early stage. Furthermore, the levels of polyamines, polyols, glycolysis, and tricarboxylic acid cycle intermediates, which changed more dramatically during the pilot process, were all higher in the pilot than in the industrial fermentation during the early stage. This indicated that the fermentation conditions of the early stage should be the focus of process management which is aimed at increasing penicillin production. Additionally, the comparative accumulations of the precursors of penicillin (valine, cysteine, and lysine) revealed that penicillin biosynthesis in the industrial process was more affected during the middle stage of fermentation. These findings provide new insights to further regulate the industrial process and improve the production of penicillin. More generally, this study attempts to address the scarcity of studies that contrast the metabolic outcomes between commercial- and pilot-scale conditions.

  18. Secondary metabolites from a marine-derived endophytic fungus Penicillium chrysogenum QEN-24S.

    PubMed

    Gao, Shu-Shan; Li, Xiao-Ming; Du, Feng-Yu; Li, Chun-Shun; Proksch, Peter; Wang, Bin-Gui

    2010-12-27

    Penicillium chrysogenum QEN-24S, an endophytic fungus isolated from an unidentified marine red algal species of the genus Laurencia, displayed inhibitory activity against the growth of pathogen Alternaria brassicae in dual culture test. Chemical investigation of this fungal strain resulted in the isolation of four new (1-3 and 5) and one known (4) secondary metabolites. Their structures were identified as two polyketide derivatives penicitides A and B (1 and 2), two glycerol derivatives 2-(2,4-dihydroxy-6-methylbenzoyl)-glycerol (3) and 1-(2,4-dihydroxy-6-methylbenzoyl)- glycerol (4), and one monoterpene derivative penicimonoterpene (5). Penicitides A and B (1 and 2) feature a unique 10-hydroxy- or 7,10-dihydroxy-5,7-dimethylundecyl moiety substituting at C-5 of the α-tetrahydropyrone ring, which is not reported previously among natural products. Compound 5 displayed potent activity against the pathogen A. brassicae, while compound 1 exhibited moderate cytotoxic activity against the human hepatocellular liver carcinoma cell line.

  19. Biodegradation of phenol in static cultures by Penicillium chrysogenum ERK1: catalytic abilities and residual phytotoxicity.

    PubMed

    Wolski, Erika A; Barrera, Viviana; Castellari, Claudia; González, Jorge F

    2012-01-01

    A phenol-degrading fungus was isolated from crop soils. Molecular characterization (using internal transcribed spacer, translation elongation factor and beta-tubulin gene sequences) and biochemical characterization allowed to identify the fungal strain as Penicillium chrysogenum Thom ERK1. Phenol degradation was tested at 25 degrees C under resting mycelium conditions at 6, 30, 60, 200, 350 and 400 mg/l of phenol as the only source of carbon and energy. The time required for complete phenol degradation increased at different initial phenol concentrations. Maximum specific degradation rate (0.89978 mg of phenol/day/mg of dry weight) was obtained at 200 mg/l. Biomass yield decreased at initial phenol concentrations above 60 mg/l. Catechol was identified as an intermediate metabolite by HPLC analysis and catechol dioxygenase activity was detected in plate assays, suggesting that phenol metabolism could occur via ortho fission of catechol. Wheat seeds were used as phytotoxicity indicators of phenol degradation products. It was found that these products were not phytotoxic for wheat but highly phytotoxic for phenol. The high specific degradation rates obtained under resting mycelium conditions are considered relevant for practical applications of this fungus in soil decontamination processes.

  20. A natural Anopheles-associated Penicillium chrysogenum enhances mosquito susceptibility to Plasmodium infection

    PubMed Central

    Angleró-Rodríguez, Yesseinia I.; Blumberg, Benjamin J.; Dong, Yuemei; Sandiford, Simone L.; Pike, Andrew; Clayton, April M.; Dimopoulos, George

    2016-01-01

    Whereas studies have extensively examined the ability of bacteria to influence Plasmodium infection in the mosquito, the tripartite interactions between non-entomopathogenic fungi, mosquitoes, and Plasmodium parasites remain largely uncharacterized. Here we report the isolation of a common mosquito-associated ascomycete fungus, Penicillium chrysogenum, from the midgut of field-caught Anopheles mosquitoes. Although the presence of Pe. chrysogenum in the Anopheles gambiae midgut does not affect mosquito survival, it renders the mosquito significantly more susceptible to Plasmodium infection through a secreted heat-stable factor. We further provide evidence that the mechanism of the fungus-mediated modulation of mosquito susceptibility to Plasmodium involves an upregulation of the insect’s ornithine decarboxylase gene, which sequesters arginine for polyamine biosynthesis. Arginine plays an important role in the mosquito’s anti-Plasmodium defense as a substrate of nitric oxide production, and its availability therefore has a direct impact on the mosquito’s susceptibility to the parasite. While this type of immunomodulatory mechanism has already been demonstrated in other host-pathogen interaction systems, this is the first report of a mosquito-associated fungus that can suppress the mosquito’s innate immune system in a way that would favor Plasmodium infection and possibly malaria transmission. PMID:27678168

  1. Effects of Penicillium chrysogenum var. halophenolicum on kraft lignin: color stabilization and cytotoxicity evaluation.

    PubMed

    Remédios, Marlene; Carvalho, Filomena A; Enguita, Francisco J; Cardoso, Carlos; Martins, Ivo C; Santos, Nuno C; Leitão, Ana Lúcia

    2016-06-01

    Wood industries and agricultural crops generate an inexhaustible supply of by-products like lignin, which constitutes an environmental problem. Increasing efforts have been done to find new applications for lignin. One of them is as a food additive, but its chemical nature makes it sensitive to browning which constitutes a major drawback for this type of lignin application. In the present study we are documenting how color stabilization of a commercial kraft lignin was achieved after the treatment with Penicillium chrysogenum var. halophenolicum. In addition the fungal capacity to remove lignin is studied together with the effect of its treatment on cytotoxicity of lignin. P. chrysogenum var. halophenolicum was able to transform lignin, ensuring its color stability for more than 24 months. Dynamic light scattering and atomic force microscopy showed that the fungus contributed to homogenize particle size and hydrodynamic properties in lignin suspensions without increase the toxicity over HeLa cells and human primary fibroblasts. These findings suggest new uses for kraft lignin after P. chrysogenum var. halophenolicum treatment providing an effective approach for improve color stability.

  2. Biodegradation of waste grease by Penicillium chrysogenum for production of fatty acid.

    PubMed

    Kumari, Arti; Ahmad, Razi; Negi, Sangeeta; Khare, Sunil Kumar

    2017-02-01

    The aim of present work was to effectively remediate grease waste by Penicillium chrysogenum. For efficient degradation, grease waste was pre-treated using various lipases, among them lipolase was the best. The pretreated grease was used as a substrate by P. chrysogenum resulting into the production of fatty acids. Process was optimized by response surface methodology (RSM) using four variables viz; FeCl2 (mM), spore concentration (spores/ml), time period (days) and amount of grease (g). The optimized conditions viz; FeCl2 1.25mM, culture amount 5×10(11)spores/ml and time period 16days led to the production of 6.6mg/g fatty acid from 10.0g of pre-treated grease mixed with 5.0g wheat bran in 10.0ml czapek-dox medium under solid state fermentation. The fermented media was extracted with hexane and subjected to GCMS analysis, which showed the presence of higher amount of palmitic acid. It was purified by crystallization method and 2.8g of palmitic acid was recovered from 1.0kg grease waste in tray fermentation.

  3. Analysis of autophagy in Penicillium chrysogenum by using starvation pads in combination with fluorescence microscopy.

    PubMed

    Scheckhuber, Christian Q

    2015-02-01

    The study of cellular quality control systems has emerged as a highly dynamic and relevant field of contemporary research. It has become clear that cells possess several lines of defense against damage to biologically relevant molecules like nucleic acids, lipids and proteins. In addition to organelle dynamics (fusion/fission/motility/inheritance) and tightly controlled protease activity, the degradation of surplus, damaged or compromised organelles by autophagy (cellular 'self-eating') has received much attention from the scientific community. The regulation of autophagy is quite complex and depends on genetic and environmental factors, many of which have so far not been elucidated. Here a novel method is presented that allows the convenient study of autophagy in the filamentous fungus Penicillium chrysogenum. It is based on growth of the fungus on so-called 'starvation pads' for stimulation of autophagy in a reproducible manner. Samples are directly assayed by microscopy and evaluated for autophagy induction / progress. The protocol presented here is not limited for use with P. chrysogenum and can be easily adapted for use in other filamentous fungi.

  4. Pyomelanin production in Penicillium chrysogenum is stimulated by L-tyrosine.

    PubMed

    Vasanthakumar, Archana; DeAraujo, Alice; Mazurek, Joy; Schilling, Michael; Mitchell, Ralph

    2015-06-01

    From a tomb in Upper Egypt we isolated a strain of Penicillium chrysogenum that was capable of producing brown pigment in vitro when grown in a minimal salts medium containing tyrosine. We present evidence that this pigment is a pyomelanin, a compound that is known to assist in the survival of some micro-organisms in adverse environments. We tested type strains of Pe. chrysogenum, which were also able to produce this pigment under similar conditions. Inhibitors of the DHN and DOPA melanin pathways were unable to inhibit the formation of the pigment. Fourier transform IR analysis indicated that this brown pigment is similar to pyomelanin. Pyrolysis-GC/MS revealed the presence of phenolic compounds. Using LC/MS, homogentisic acid, the monomeric precursor of pyomelanin, was detected in supernatants of Pe. chrysogenum cultures growing in tyrosine medium but not in cultures lacking tyrosine. Partial regions of the genes encoding two enzymes in the homogentisic acid pathway of tyrosine degradation were amplified. Data from reverse-transcription PCR demonstrated that hmgA transcription was increased in cultures grown in tyrosine medium, suggesting that tyrosine induced the transcription.

  5. A branched biosynthetic pathway is involved in production of roquefortine and related compounds in Penicillium chrysogenum.

    PubMed

    Ali, Hazrat; Ries, Marco I; Nijland, Jeroen G; Lankhorst, Peter P; Hankemeier, Thomas; Bovenberg, Roel A L; Vreeken, Rob J; Driessen, Arnold J M

    2013-01-01

    Profiling and structural elucidation of secondary metabolites produced by the filamentous fungus Penicillium chrysogenum and derived deletion strains were used to identify the various metabolites and enzymatic steps belonging to the roquefortine/meleagrin pathway. Major abundant metabolites of this pathway were identified as histidyltryptophanyldiketopiperazine (HTD), dehydrohistidyltryptophanyldi-ketopiperazine (DHTD), roquefortine D, roquefortine C, glandicoline A, glandicoline B and meleagrin. Specific genes could be assigned to each enzymatic reaction step. The nonribosomal peptide synthetase RoqA accepts L-histidine and L-tryptophan as substrates leading to the production of the diketopiperazine HTD. DHTD, previously suggested to be a degradation product of roquefortine C, was found to be derived from HTD involving the cytochrome P450 oxidoreductase RoqR. The dimethylallyltryptophan synthetase RoqD prenylates both HTD and DHTD yielding directly the products roquefortine D and roquefortine C without the synthesis of a previously suggested intermediate and the involvement of RoqM. This leads to a branch in the otherwise linear pathway. Roquefortine C is subsequently converted into glandicoline B with glandicoline A as intermediates, involving two monooxygenases (RoqM and RoqO) which were mixed up in an earlier attempt to elucidate the biosynthetic pathway. Eventually, meleagrin is produced from glandicoline B involving a methyltransferase (RoqN). It is concluded that roquefortine C and meleagrin are derived from a branched biosynthetic pathway.

  6. Inactivation of conidia of Penicillium chrysogenum, P. digitatum and P. italicum by ethanol solutions and vapours.

    PubMed

    Dao, Thien; Bensoussan, Maurice; Gervais, Patrick; Dantigny, Philippe

    2008-02-29

    A fractional factorial design, 2(5-1) experiments, was used for assessing the influence of 5 factors: water activity, aw [0.7, 0.9], temperature, T ( degrees C) [10, 30], mode of application, A [liquid, vapour], ethanol concentration, E (% w/w) [5, 10] and time, t (d) [1, 4] on the inactivation of spores of Penicillium chrysogenum, P. digitatum and P. italicum. Survival was determined by germination at optimal conditions within 3d. The experimental response was log (N 0/Nt), where N 0 and Nt (spore ml(-1)) the concentrations of viable spores at t=0 and t respectively. By a decreasing order of sensitivity to ethanol, moulds were ranked as followed: P. digitatum, P. italicum and P. chrysogenum. A greater inactivation for P. digitatum, P. italicum, that were the most sensitive moulds to ethanol, was obtained by applying vapour rather than ethanol solution. The order of significance of the main factors depended upon the mould. The key factor for explaining inactivation of P. chrysogenum was water activity. But, temperature was the main factor for explaining inactivation of P. digitatum and P. italicum. In the more drastic conditions, (i.e., 0.7 aw, 30 degrees C, 10% w/w ethanol), all spores were inactivated by applying liquid solution for 4d.

  7. Simultaneous utilization of glucose and gluconate in Penicillium chrysogenum during overflow metabolism.

    PubMed

    Schmitz, Katja; Peter, Vivien; Meinert, Sabine; Kornfeld, Georg; Hardiman, Timo; Wiechert, Wolfgang; Noack, Stephan

    2013-12-01

    The filamentous fungus Penicillium chrysogenum is one of the most important production organism for β-lactam antibiotics, especially penicillin. A specific feature of P. chrysogenum is the formation of gluconate as the primary overflow metabolite under non-limiting growth on glucose. Gluconate can be formed extracellularly by the enzyme glucose oxidase (GOD) that shows high activities under glucose excess conditions. Currently, it is assumed that under these conditions glucose is the preferred carbon substrate for P. chrysogenum and gluconate consumption first starts after glucose becomes limiting. Here, we specifically address this hypothesis by combining batch cultivation experiments on defined glucose media, time-dependent GOD activity measurements, and (13)C-tracer studies. Our data prove that both substrates are metabolized simultaneously independent from the actual glucose concentration and therefore suggest that no distinct mechanism of carbon catabolite repression exists for gluconate in P. chrysogenum. Moreover, gluconate consumption does not interfere with penicillin V production by repression of the penicillin genes. Finally, by following a model-driven approach the specific uptake rates for glucose and gluconate were quantified and found to be significantly higher for gluconate. In summary, our results show that P. chrysogenum metabolizes gluconate directly and at high rates making it an interesting alternative carbon source for production purposes.

  8. Purification and characterization of a novel antifungal protein secreted by Penicillium chrysogenum from an Arctic sediment.

    PubMed

    Chen, Zhiteng; Ao, Jingqun; Yang, Wenchuan; Jiao, Liping; Zheng, Tianling; Chen, Xinhua

    2013-12-01

    A fungal strain, Penicillium chrysogenum A096, was isolated from an Arctic sediment sample. Its culture supernatant inhibited mycelial growth of some plant pathogenic fungi. After saturation of P. chrysogenum A096 culture supernatant with ammonium sulfate and ion exchange chromatography, a novel antifungal protein (Pc-Arctin) was purified and identified by matrix assisted laser desorption ionization-time of flight-time of flight-mass spectrometry (MALDI-TOF-TOF-MS). The gene encoding for Pc-Arctin consisting of 195 nucleotides was cloned from P. chrysogenum A096 to confirm the mass spectrometry result. Pc-Arctin displays antifungal activity against Paecilomyces variotii, Alternaria longipes, and Trichoderma viride at minimum inhibitory concentrations (MIC) of 24, 48, and 192 ng/disc, respectively. Pc-Arctin was most sensitive to proteinase K and then to trypsin but insensitive to papain. Pc-Arctin possesses high thermostability and cannot be antagonized by common surfactants, except for sodium dodecyl sulfate (SDS). Divalent ions, such as Mn(2+), Mg(2+), and Zn(2+), inhibited the antifungal activity of Pc-Arctin. Hemagglutination assays showed that Pc-Arctin had no hemagglutinating or hemolytic activity against red blood cells (RBC) from rabbits, rats, and guinea pigs. Therefore, Pc-Arctin from Arctic P. chrysogenum may represent a novel antifungal protein with potential for application in controlling plant pathogenic fungal infection.

  9. Cytosolic NADPH balancing in Penicillium chrysogenum cultivated on mixtures of glucose and ethanol.

    PubMed

    Zhao, Zheng; Kuijvenhoven, Karel; van Gulik, Walter M; Heijnen, Joseph J; van Winden, Wouter A; Verheijen, Peter J T

    2011-01-01

    The in vivo flux through the oxidative branch of the pentose phosphate pathway (oxPPP) in Penicillium chrysogenum was determined during growth in glucose/ethanol carbon-limited chemostat cultures, at the same growth rate. Non-stationary (13)C flux analysis was used to measure the oxPPP flux. A nearly constant oxPPP flux was found for all glucose/ethanol ratios studied. This indicates that the cytosolic NADPH supply is independent of the amount of assimilated ethanol. The cofactor assignment in the model of van Gulik et al. (Biotechnol Bioeng 68(6):602-618, 2000) was supported using the published genome annotation of P. chrysogenum. Metabolic flux analysis showed that NADPH requirements in the cytosol remain nearly the same in these experiments due to constant biomass growth. Based on the cytosolic NADPH balance, it is known that the cytosolic aldehyde dehydrogenase in P. chrysogenum is NAD(+) dependent. Metabolic modeling shows that changing the NAD(+)-aldehyde dehydrogenase to NADP(+)-aldehyde dehydrogenase can increase the penicillin yield on substrate.

  10. Bioleaching of heavy metals from a contaminated soil using indigenous Penicillium chrysogenum strain F1.

    PubMed

    Deng, Xinhui; Chai, Liyuan; Yang, Zhihui; Tang, Chongjian; Tong, Haixia; Yuan, Pingfu

    2012-09-30

    Bioleaching of heavy metals from contaminated soil using Penicillium chrysogenum strain F1 was investigated. Batch experiments were performed to compare leaching efficiencies of heavy metals between one-step and two-step processes and to determine the transformation of heavy metal fractions before and after bioleaching. The results showed that two-step process had higher leaching efficiencies of heavy metals than one-step process. When the mass ratio of soil to culture medium containing P. chrysogenum strain F1 was 5% (w/v), 50%, 35%, 9% and 40% of Cd, Cu, Pb and Zn were removed in one-step process, respectively. The two-step process had higher removals of 63% Cd, 56% Cu, 14% Pb and 54% Zn as compared with one-step process. The results of the sequential extraction showed that the metals remaining in the soil were mainly bonded in stable fractions after bioleaching. The results of TEM and SEM showed that during bioleaching process, although the mycelium of P. chrysogenum was broken into fragments, no damage was obviously observed on the surface of the living cell except for thinner cell wall, smaller vacuoles and concentrated cytoplasm. The result implied that P. chrysogenum strain F1 can be used to remove heavy metals from polluted soil.

  11. Biochemical Characterization and Overexpression of an Endo-rhamnogalacturonan Lyase from Penicillium chrysogenum.

    PubMed

    Iwai, Marin; Yamada, Hiroyuki; Ikemoto, Takeshi; Matsumoto, Shotaro; Fujiwara, Daisuke; Takenaka, Shigeo; Sakamoto, Tatsuji

    2015-06-01

    Rhamnogalacturonan lyase (PcRGL4A) was purified from the culture supernatant of Penicillium chrysogenum 31B. PcRGL4A optimal activity occurred between pH 7-8 and at 40 °C. Conserved Domain Search analysis identified PcRGL4A as a member of Polysaccharide Lyase family 4. PcRGL4A contains two conserved catalytic and four conserved substrate-binding residues as determined by X-ray crystallography of the Aspergillus aculeatus RG lyase. Recombinant PcRGL4A (rPcRGL4A) expressed in Escherichia coli demonstrated specific activity against rhamnogalacturonan (RG) but not homogalacturonan. Analysis of the RG reaction products by high-performance anion-exchange chromatography revealed that rPcRGL4A cleaved the substrate in an endo-manner and that the major final product was an RG tetrasaccharide with 4-deoxy-4,5-unsaturated galacturonic acid at the nonreducing end. Based on these results, PcRGL4A was classified as an endo-acting RG lyase (EC 4.2.2.23). Divalent cations were not essential for the enzymatic activity of rPcRGL4A, but addition of calcium ions to the reaction mixture increased enzymatic activity. rPcRGL4A demonstrated a preference for RG lacking galactose decoration.

  12. Evidence for Dicer-dependent RNA interference in the industrial penicillin producer Penicillium chrysogenum.

    PubMed

    Janus, Danielle; Hoff, Birgit; Kück, Ulrich

    2009-12-01

    RNA interference (RNAi) is a sequence-specific post-transcriptional gene silencing system that downregulates target gene expression. Here, we provide several lines of evidence for RNA silencing in the industrial beta-lactam antibiotic producer Penicillium chrysogenum using the DsRed reporter gene under the control of the constitutive trpC promoter or the inducible xylP promoter. The functional RNAi system was verified by detection of siRNAs that hybridized exclusively with gene-specific (32)P-labelled RNA probes. Moreover, when RNAi was used to silence the endogenous PcbrlA morphogene that controls conidiophore development, a dramatic reduction in the formation of conidiospores was observed in 47 % of the corresponding transformants. Evidence that RNAi in P. chrysogenum is dependent on a Dicer peptide was provided with a strain lacking Pcdcl2. In the DeltaPcdcl2 background, silencing of the PcbrlA gene was tested. None of the transformants analysed showed a developmental defect. The applicability of the RNAi system in P. chrysogenum was finally demonstrated by silencing the Pcku70 gene to increase homologous recombination frequency. This led to the generation of single and double knockout mutants.

  13. Matching the proteome to the genome: the microbody of penicillin-producing Penicillium chrysogenum cells.

    PubMed

    Kiel, Jan A K W; van den Berg, Marco A; Fusetti, Fabrizia; Poolman, Bert; Bovenberg, Roel A L; Veenhuis, Marten; van der Klei, Ida J

    2009-05-01

    In the filamentous fungus Penicillium chrysogenum, microbodies are essential for penicillin biosynthesis. To better understand the role of these organelles in antibiotics production, we determined the matrix enzyme contents of P. chrysogenum microbodies. Using a novel in silico approach, we first obtained a catalogue of 200 P. chrysogenum proteins with putative microbody targeting signals (PTSs). This included two orthologs of proteins involved in cephalosporin biosynthesis, which we demonstrate to be bona fide microbody matrix constituents. Subsequently, we performed a proteomics based inventory of P. chrysogenum microbody matrix proteins using nano-LC-MS/MS analysis. We identified 89 microbody proteins, 79 with a PTS, including the two known microbody-borne penicillin biosynthesis enzymes, isopenicillin N:acyl CoA acyltransferase and phenylacetyl-CoA ligase. Comparative analysis revealed that 69 out of 79 PTS proteins identified experimentally were in the reference list. A prominent microbody protein was identified as a novel fumarate reductase-cytochrome b5 fusion protein, which contains an internal PTS2 between the two functional domains. We show that this protein indeed localizes to P. chrysogenum microbodies.

  14. Engineering of Penicillium chrysogenum for fermentative production of a novel carbamoylated cephem antibiotic precursor.

    PubMed

    Harris, Diana M; Westerlaken, Ilja; Schipper, Dick; van der Krogt, Zita A; Gombert, Andreas K; Sutherland, John; Raamsdonk, Leonie M; van den Berg, Marco A; Bovenberg, Roel A L; Pronk, Jack T; Daran, Jean-Marc

    2009-03-01

    Penicillium chrysogenum was successfully engineered to produce a novel carbamoylated cephalosporin that can be used as a synthon for semi-synthetic cephalosporins. To this end, genes for Acremonium chrysogenum expandase/hydroxylase and Streptomyces clavuligerus carbamoyltransferase were expressed in a penicillinG high-producing strain of P.chrysogenum. Growth of the engineered strain in the presence of adipic acid resulted in production of adipoyl-7-amino-3-carbamoyloxymethyl-3-cephem-4-carboxylic acid (ad7-ACCCA) and of several adipoylated pathway intermediates. A combinatorial chemostat-based transcriptome study, in which the ad7-ACCCA-producing strain and a strain lacking key genes in beta-lactam synthesis were grown in the presence and absence of adipic acid, enabled the dissection of transcriptional responses to adipic acid per se and to ad7-ACCCA production. Transcriptome analysis revealed that adipate catabolism in P.chrysogenum occurs via beta-oxidation and enabled the identification of putative genes for enzymes involved in mitochondrial and peroxisomal beta-oxidation pathways. Several of the genes that showed a specifically altered transcript level in ad7-ACCCA-producing cultures were previously implicated in oxidative stress responses.

  15. Cloning and characterization of a novel CoA-ligase gene from Penicillium chrysogenum.

    PubMed

    Yu, Zhou-Liang; Liu, Jing; Wang, Fu-Qiang; Dai, Meng; Zhao, Bao-Hua; He, Jian-Gong; Zhang, Hua

    2011-05-01

    A novel phenylacetic acid (PAA)-induced CoA-ligase-encoding gene, designated as phlC, has been cloned from penicillin-producing fungus Penicillium chrysogenum. The open reading frame of phlC cDNA was 1671 bp and encoded a 556 amino acid residues protein with the consensus AMP binding site and a peroxisomal targeting signal 1 on its C terminus. The deduced amino acid sequence showed 37% and 38% identity with characterized P. chrysogenum Phl and PhlB protein, respectively. Functional recombinant PhlC protein was overexpressed in Escherichia coli. The purified recombinant enzyme was capable to convert PAA into its corresponding CoA ester with a specific activity of 129.5 ± 3.026 pmol/min per mg protein. Similar to Phl and PhlB, PhlC displayed broad substrate spectrum and showed higher activities to medium- and long-chain fatty acids. The catalytic properties of PhlC have been determined and compared to those of Phl and PhlB.

  16. Comparison of the secondary metabolites in Penicillium chrysogenum between pilot and industrial penicillin G fermentations.

    PubMed

    Cao, Ying-Xiu; Qiao, Bin; Lu, Hua; Chen, Yao; Yuan, Ying-Jin

    2011-02-01

    The disparity of secondary metabolites in Penicillium chrysogenum between two scales of penicillin G fermentation (50 L as pilot process and 150,000 L as industrial one) was investigated by ion-pair reversed-phase liquid chromatography tandemed with hybrid quadrupole time-of-flight mass spectrometry. In industrial process, the pools of intracellular L-α-aminoadipyl-L-cysteinyl-D-valine (LLD-ACV) and isopenicillin N (IPN) were remarkably less than that in the pilot one, which indicated that the productivity of penicillin G might be higher in the large scale of fermentation. This conclusion was supported by the higher intracellular penicillin G concentration as well as its higher yield per unit biomass in industrial cultivation. The different changing tendencies of IPN, 6-aminopenicillanic acid and 6-oxopiperide-2-carboxylic acid between two processes also suggested the same conclusion. The higher content of intracellular LLD-ACV in pilot process lead to a similarly higher concentration of bis-δ-(L-α-aminoadipyl)-L-cysteinyl-D-valine, which had an inhibitory effect on ACV synthetase and also subdued the activity of IPN synthetase. The interconversion of secondary metabolites and the influence they put on enzymes would intensify the discrepancy between two fermentations more largely. These findings provided new insight into the changes and regulation of secondary metabolites in P. chrysogenum under different fermentation sizes.

  17. Increased penicillin production in Penicillium chrysogenum production strains via balanced overexpression of isopenicillin N acyltransferase.

    PubMed

    Weber, Stefan S; Polli, Fabiola; Boer, Rémon; Bovenberg, Roel A L; Driessen, Arnold J M

    2012-10-01

    Intense classical strain improvement has yielded industrial Penicillium chrysogenum strains that produce high titers of penicillin. These strains contain multiple copies of the penicillin biosynthesis cluster encoding the three key enzymes: δ-(l-α-aminoadipyl)-L-cysteinyl-D-valine synthetase (ACVS), isopenicillin N synthase (IPNS), and isopenicillin N acyltransferase (IAT). The phenylacetic acid coenzyme A (CoA) ligase (PCL) gene encoding the enzyme responsible for the activation of the side chain precursor phenylacetic acid is localized elsewhere in the genome in a single copy. Since the protein level of IAT already saturates at low cluster copy numbers, IAT might catalyze a limiting step in high-yielding strains. Here, we show that penicillin production in high-yielding strains can be further improved by the overexpression of IAT while at very high levels of IAT the precursor 6-aminopenicillic acid (6-APA) accumulates. Overproduction of PCL only marginally stimulates penicillin production. These data demonstrate that in high-yielding strains IAT is the limiting factor and that this limitation can be alleviated by a balanced overproduction of this enzyme.

  18. High-resolution structure of exo-arabinanase from Penicillium chrysogenum.

    PubMed

    Sogabe, Yuri; Kitatani, Tomoya; Yamaguchi, Asako; Kinoshita, Takayoshi; Adachi, Hiroaki; Takano, Kazufumi; Inoue, Tsuyoshi; Mori, Yusuke; Matsumura, Hiroyoshi; Sakamoto, Tatsuji; Tada, Toshiji

    2011-05-01

    Arabinanase Abnx from Penicillium chrysogenum 31B, which belongs to the GH93 family, releases arabinobiose from the nonreducing terminus of α-1,5-L-arabinan, which is distributed in the primary cell walls of higher plants. Crystal structures of Abnx and of its complex with arabinobiose were determined at the high resolutions of 1.14 Å to an R(work) of 10.7% (R(free) = 12.8%) and 1.04 Å to an R(work) of 10.4% (R(free) = 12.5%). Abnx has a six-bladed β-propeller fold with a typical ring-closure mode called `Velcro', in which the last four-stranded β-sheet is completed by the incorporation of a strand from the N-terminus. Catalytic residues which act as a nucleophile and an acid/base were proposed from the structures and confirmed by site-directed mutagenesis. The substrate-binding groove is enclosed at one end by two residues, Glu64 and Tyr66, which contribute to the recognition of the nonreducing chain end of the polysaccharide. A comparison with the related enzyme Arb93A which has a quite similar overall structure suggested that Abnx has different mechanisms to funnel substrates to the active site and/or to stabilize the transition state.

  19. Nonlinear biosynthetic gene cluster dose effect on penicillin production by Penicillium chrysogenum.

    PubMed

    Nijland, Jeroen G; Ebbendorf, Bjorg; Woszczynska, Marta; Boer, Rémon; Bovenberg, Roel A L; Driessen, Arnold J M

    2010-11-01

    Industrial penicillin production levels by the filamentous fungus Penicillium chrysogenum increased dramatically by classical strain improvement. High-yielding strains contain multiple copies of the penicillin biosynthetic gene cluster that encodes three key enzymes of the β-lactam biosynthetic pathway. We have analyzed the gene cluster dose effect on penicillin production using the high-yielding P. chrysogenum strain DS17690 that was cured from its native clusters. The amount of penicillin V produced increased with the penicillin biosynthetic gene cluster number but was saturated at high copy numbers. Likewise, transcript levels of the biosynthetic genes pcbAB [δ-(l-α-aminoadipyl)-l-cysteinyl-d-valine synthetase], pcbC (isopenicillin N synthase), and penDE (acyltransferase) correlated with the cluster copy number. Remarkably, the protein level of acyltransferase, which localizes to peroxisomes, was saturated already at low cluster copy numbers. At higher copy numbers, intracellular levels of isopenicillin N increased, suggesting that the acyltransferase reaction presents a limiting step at a high gene dose. Since the number and appearance of the peroxisomes did not change significantly with the gene cluster copy number, we conclude that the acyltransferase activity is limiting for penicillin biosynthesis at high biosynthetic gene cluster copy numbers. These results suggest that at a high penicillin production level, productivity is limited by the peroxisomal acyltransferase import activity and/or the availability of coenzyme A (CoA)-activated side chains.

  20. UVA Photoactivation of Harmol Enhances Its Antifungal Activity against the Phytopathogens Penicillium digitatum and Botrytis cinerea

    PubMed Central

    Olmedo, Gabriela M.; Cerioni, Luciana; González, María M.; Cabrerizo, Franco M.; Volentini, Sabrina I.; Rapisarda, Viviana A.

    2017-01-01

    Phytopathogenic fungi responsible for post-harvest diseases on fruit and vegetables cause important economic losses. We have previously reported that harmol (1-methyl-9H-pyrido[3,4-b]indol-7-ol) is active against the causal agents of green and gray molds Penicillium digitatum and Botrytis cinerea, respectively. Here, antifungal activity of harmol was characterized in terms of pH dependency and conidial targets; also photodynamic effects of UVA irradiation on the antimicrobial action were evaluated. Harmol was able to inhibit the growth of both post-harvest fungal disease agents only in acidic conditions (pH 5), when it was found in its protonated form. Conidia treated with harmol exhibited membrane integrity loss, cell wall disruption, and cytoplasm disorganization. All these deleterious effects were more evident for B. cinerea in comparison to P. digitatum. When conidial suspensions were irradiated with UVA in the presence of harmol, antimicrobial activity against both pathogens was enhanced, compared to non-irradiated conditions. B. cinerea exhibited a high intracellular production of reactive oxygen species (ROS) when was incubated with harmol in irradiated and non-irradiated treatments. P. digitatum showed a significant increase in ROS accumulation only when treated with photoexcited harmol. The present work contributes to unravel the antifungal activity of harmol and its photoexcited counterpart against phytopathogenic conidia, focusing on ROS accumulation which could account for damage on different cellular targets. PMID:28326067

  1. Organic Acid Excretion in Penicillium ochrochloron Increases with Ambient pH

    PubMed Central

    Vrabl, Pamela; Fuchs, Viktoria; Pichler, Barbara; Schinagl, Christoph W.; Burgstaller, Wolfgang

    2012-01-01

    Despite being of high biotechnological relevance, many aspects of organic acid excretion in filamentous fungi like the influence of ambient pH are still insufficiently understood. While the excretion of an individual organic acid may peak at a certain pH value, the few available studies investigating a broader range of organic acids indicate that total organic acid excretion rises with increasing external pH. We hypothesized that this phenomenon might be a general response of filamentous fungi to increased ambient pH. If this is the case, the observation should be widely independent of the organism, growth conditions, or experimental design and might therefore be a crucial key point in understanding the function and mechanisms of organic acid excretion in filamentous fungi. In this study we explored this hypothesis using ammonium-limited chemostat cultivations (pH 2–7), and ammonium or phosphate-limited bioreactor batch cultivations (pH 5 and 7). Two strains of Penicillium ochrochloron were investigated differing in the spectrum of excreted organic acids. Confirming our hypothesis, the main result demonstrated that organic acid excretion in P. ochrochloron was enhanced at high external pH levels compared to low pH levels independent of the tested strain, nutrient limitation, and cultivation method. We discuss these findings against the background of three hypotheses explaining organic acid excretion in filamentous fungi, i.e., overflow metabolism, charge balance, and aggressive acidification hypothesis. PMID:22493592

  2. Penicillium species-induced granuloma in a cat resulting in chronic lower urinary tract disease.

    PubMed

    Soonthornsit, Jeerawat; Banlunara, Wijit; Niyomthum, Waree; Pusoonthornthum, Rosama

    2013-12-01

    A 5-year-old, female neutered Persian cat was admitted to the Small Animal Hospital (Chulalongkorn University, Bangkok, Thailand) with clinical signs of dysuria, haematuria and partial urethral obstruction that had manifested over several months. The animal also had hyperkalaemia and severe azotaemia at the time of presentation. Urinalysis showed haematuria, pyuria and the presence of several transitional cells. In addition, ultrasonography demonstrated an extraluminal mass between the neck of urinary bladder and the colon. Fine-needle aspiration of the mass revealed a fungal form with branching and septate hyphae. Consequently, itraconazole treatment was prescribed and clinical signs of improvement were seen after 7 days. However, 1 month later, the cat died of acute anaemia. Necropsy revealed the presence of extraluminal multifocal fungal granuloma at the neck of the urinary bladder, and contracted kidneys. Histopathological analysis of the fungal granuloma was found to be composed of branching, septate hyphal fungi together with inflammatory cells. Subsequent fungal culture and identification revealed this to be a species of Penicillium.

  3. Characterization of the ice nucleation activity of an airborne Penicillium species

    NASA Astrophysics Data System (ADS)

    Yordanova, Petya; Hill, Thomas C. J.; Pummer, Bernhard G.; Franc, Gary D.; Pöschl, Ulrich; Fröhlich-Nowoisky, Janine

    2016-04-01

    Microorganisms are ubiquitous both on and above the Earth. Several bacterial and fungal spe-cies are the focus of atmospheric studies due to their ability to trigger ice formation at high subzero temperatures. Thus, they have potential to modify cloud albedo, lifetime and precipita-tion, and ultimately the hydrological cycle. Several fungal strains have already been identified as possessing ice nucleation (IN) activity, and recent studies have shown that IN active fungi are present in the cultivable community of air and soil samples [1, 2]. However, the abundance, diversity, and sources of fungal ice nuclei in the atmosphere are still poorly characterized. In this study, fungal colonies obtained from air samples were screened for IN activity in the droplet-freezing assay described in Fröhlich-Nowoisky et al., 2015 [2]. Out of 128 tested iso-lates, two were found to catalyze ice formation at temperatures up to -4°C. By DNA analysis, both isolates were classified as Penicillium spp. The freezing activity of both was further char-acterized after different filtration, heat, and enzymatic treatments in the temperature range from -4°C to -15°C. Preliminary results show that a proteinaceous compound is responsible for the IN activity. Furthermore, ongoing experiments indicate that the activity is associated only with the hyphae. [1] Huffman, et al. (2013): Atmos. Chem. Phys., 13, 6151-6164. [2] Fröhlich-Nowoisky et al. (2015): Biogeosciences, 12: 1057-1071.

  4. A gene encoding a new cold-active lipase from an Antarctic isolate of Penicillium expansum.

    PubMed

    Mohammed, Suja; Te'o, Junior; Nevalainen, Helena

    2013-08-01

    Cold-active lipases are of significant interest as biocatalysts in industrial processes. We have identified a lipase that displayed activity towards long carbon-chain-p-nitrophenyl substrates (C12-C18) at 25 °C from the culture supernatant of an Antarctic Penicillium expansum strain assigned P. expansum SM3. Zymography revealed a protein band of around 30 kDa with activity towards olive oil. DNA fragments of a lipase gene designated as lipPE were isolated from the genomic DNA of P. expansum SM3 by genomic walking PCR. Subsequently, the complete genomic lipPE gene was amplified using gene-specific primers designed from the 5'- and 3'-regions. Reverse transcription PCR was used to amplify the lipPE cDNA. The deduced amino acid sequence consisted of 285 residues that included a predicted signal peptide. Three peptides identified by LC/MS/MS analysis of the proteins in the culture supernatant of P. expansum were also present in the deduced amino acid sequence of the lipPE gene suggesting that this gene encoded the lipase identified by initial zymogram activity analysis. Full analysis of the nucleotide and the deduced amino acid sequences indicated that the lipPE gene encodes a novel P. expansum lipase. The lipPE gene was expressed in E. coli for further characterization of the enzyme with a view of assessing its suitability for industrial applications.

  5. Agrobacterium tumefaciens-mediated transformation of Penicillium expansum PE-12 and its application in molecular breeding.

    PubMed

    Zhang, Tian; Qi, Zhen; Wang, Yueyue; Zhang, Fangyuan; Li, Renyong; Yu, Qingsheng; Chen, Xiangbin; Wang, Huojun; Xiong, Xin; Tang, Kexuan

    2013-03-30

    Lipase produced by Penicillium expansum is widely used in laundry detergent and leather industry; however, the absence of an efficient transformation technology sets a major obstacle for further enhancement of its lipase productivity through advanced gene engineering. In this work, Agrobacterium tumefaciens-mediated transformation (ATMT) was investigated for P. expansum PE-12 transformation, using hygromycin phosphotransferase (hph) as a selectable marker gene. As a result, we revealed that the frequency of transformation surpassed 100 transformants/10(5)condida, most of the integrated T-DNA appeared as a single copy at a random position in chromosomal DNA, and all the transformants showed mitotic stability. Facilitated by this newly established method, for the first time, P. expansum PE-12 was genetically engineered to improve the lipase yield, through a homologous expression vector carrying the endogenous lipase gene (PEL) driven by the strong constitutive promoter of the glyceraldehydes-3-phosphate dehydrogenase gene (gpdA) from Aspergillus nidulans. The highest expression level of the engineered strain reached up to 1700 U/mL, nearly 2-fold of the original industrial strain (900 U/mL). Our reproducible ATMT system has not only revealed the great potential of homologous expression-directed genetic engineering, which is more efficient and specific compared to traditional mutagenesis, but also provided new possibilities and perspectives for any other practical applications of P. expansum-related genetic engineering in the future.

  6. Control of postharvest fungal pathogens by antifungal compounds from Penicillium expansum.

    PubMed

    Rouissi, Wafa; Ugolini, Luisa; Martini, Camilla; Lazzeri, Luca; Mari, Marta

    2013-11-01

    The fungicidal effects of secondary metabolites produced by a strain of Penicillium expansum (R82) in culture filtrate and in a double petri dish assay were tested against one isolate each of Botrytis cinerea, Colletotrichum acutatum, and Monilinia laxa and six isolates of P. expansum, revealing inhibitory activity against every pathogen tested. The characterization of volatile organic compounds released by the R82 strain was performed by solid-phase microextraction-gas chromatographic techniques, and several compounds were detected, one of them identified as phenethyl alcohol (PEA). Synthetic PEA, tested in vitro on fungal pathogens, showed strong inhibition at a concentration of 1,230 μg/ml of airspace, and mycelium appeared more sensitive than conidia; nevertheless, at the concentration naturally emitted by the fungus (0.726 ± 0.16 m g/ml), commercial PEA did not show any antifungal activity. Therefore, a combined effect between different volatile organic compounds produced collectively by R82 can be hypothesized. This aspect suggests further investigation into the possibility of exploiting R82 as a nonchemical alternative in the control of some plant pathogenic fungi.

  7. Antifungal activity of zinc oxide nanoparticles against Botrytis cinerea and Penicillium expansum.

    PubMed

    He, Lili; Liu, Yang; Mustapha, Azlin; Lin, Mengshi

    2011-03-20

    Antifungal activities of zinc oxide nanoparticles (ZnO NPs) and their mode of action against two postharvest pathogenic fungi (Botrytis cinerea and Penicillium expansum) were investigated in this study. ZnO NPs with sizes of 70 ± 15 nm and concentrations of 0, 3, 6 and 12 mmol l(-1) were used. Traditional microbiological plating, scanning electron microscopy (SEM), and Raman spectroscopy were used to study antifungal activities of ZnO NPs and to characterize the changes in morphology and cellular compositions of fungal hyphae treated with ZnO NPs. Results show that ZnO NPs at concentrations greater than 3 mmol l(-1) can significantly inhibit the growth of B. cinerea and P. expansum. P. expansum was more sensitive to the treatment with ZnO NPs than B. cinerea. SEM images and Raman spectra indicate two different antifungal activities of ZnO NPs against B. cinerea and P. expansum. ZnO NPs inhibited the growth of B. cinerea by affecting cellular functions, which caused deformation in fungal hyphae. In comparison, ZnO NPs prevented the development of conidiophores and conidia of P. expansum, which eventually led to the death of fungal hyphae. These results suggest that ZnO NPs could be used as an effective fungicide in agricultural and food safety applications.

  8. Effect of inoculation with Penicillium expansum on the microbial community and maturity of compost.

    PubMed

    Wang, Hong-yuan; Fan, Bing-quan; Hu, Qing-xiu; Yin, Zhong-wei

    2011-12-01

    Compost prepared from wheat straw and cattle/chicken mature was inoculated with the lignocellulolytic fungus, Penicillium expansum. Compared to uninoculated compost, the inoculated compost exhibited a 150% higher germination index, more than 1.2 g kg(-1)-dw of changes in NH(4)(+)-N concentrations, a ca. 12.0% higher humus content and a lignocellulose degradation that proceeded 57.5% faster. Culture-based determinations of microbial populations demonstrated that aerobic heterotrophic bacteria and fungi were about 1-2 orders of magnitude higher in inoculated than in uninoculated compost. The number of ammonifying, ammonium-oxidizing, nitrite-oxidizing, denitrifying bacteria and cellulose-decomposing bacteria was 6.1-9.0 log(10) CFU g(-1)-dw, 1.2-4.3 log(10) MPN g(-1)-dw, 3.5-6.8 log(10) MPN g(-1)-dw, 3.58-4.34 log(10) MPN g(-1)-dw, 1.4-3.8 log(10)MPN g(-1)-dw, and 4.2-8.8 log(10) CFU g(-1)-dw higher in the compost inoculated with P. expansum.

  9. Patulin is a cultivar-dependent aggressiveness factor favouring the colonization of apples by Penicillium expansum.

    PubMed

    Snini, Selma P; Tannous, Joanna; Heuillard, Pauline; Bailly, Sylviane; Lippi, Yannick; Zehraoui, Enric; Barreau, Christian; Oswald, Isabelle P; Puel, Olivier

    2016-08-01

    The blue mould decay of apples is caused by Penicillium expansum and is associated with contamination by patulin, a worldwide regulated mycotoxin. Recently, a cluster of 15 genes (patA-patO) involved in patulin biosynthesis was identified in P. expansum. blast analysis revealed that patL encodes a Cys6 zinc finger regulatory factor. The deletion of patL caused a drastic decrease in the expression of all pat genes, leading to an absence of patulin production. Pathogenicity studies performed on 13 apple varieties indicated that the PeΔpatL strain could still infect apples, but the intensity of symptoms was weaker compared with the wild-type strain. A lower growth rate was observed in the PeΔpatL strain when this strain was grown on nine of the 13 apple varieties tested. In the complemented PeΔpatL:patL strain, the ability to grow normally in apple and the production of patulin were restored. Our results clearly demonstrate that patulin is not indispensable in the initiation of the disease, but acts as a cultivar-dependent aggressiveness factor for P. expansum. This conclusion was strengthened by the fact that the addition of patulin to apple infected by the PeΔpatL mutant restored the normal fungal colonization in apple.

  10. Intraspecific variability of growth and patulin production of 79 Penicillium expansum isolates at two temperatures.

    PubMed

    Garcia, Daiana; Ramos, Antonio J; Sanchis, Vicente; Marín, Sonia

    2011-12-02

    Penicillium expansum is the main species responsible for patulin production in apples and pears. Generally, fruit is stored at suboptimal conditions for mould growth and this situation could influence on the intra-species variability in both capability for growth and mycotoxin production. The aim of this research was to assess the impact of suboptimal environmental conditions on the intra-specific variability of P. expansum growth and patulin production using seventy nine isolates of this mould. Petri dishes with Apple Concentrate Agar Medium (ACAM) were inoculated centrally and incubated at two temperatures, one near optimal (20 °C) and the other representative of suboptimal cold storage (1 °C). For each condition, 10 Petri dishes were inoculated, and colony growth and patulin production was measured over time. The Kruskal-Wallis test revealed significant differences among growth rate (μ) and lag phase (λ) within the seventy nine assayed isolates. Coefficients of variation revealed a wider dispersion of μ (mm/day) and λ (days) at 1 °C compared with 20 °C. There were significant differences (p<0.05) among patulin levels (ng/mm²) for the different conditions, values being lower at the lower temperature. Coefficients of variation revealed a wider dispersion of mycotoxin production at 1 °C. In order to address the strain variability in growth initiation and prove the well-established notion of reducing patulin production in foods by preventing fungal growth, a greater number of strains should be included.

  11. Substitution of Val72 residue alters the enantioselectivity and activity of Penicillium expansum lipase.

    PubMed

    Tang, Lianghua; Su, Min; Zhu, Ling; Chi, Liying; Zhang, Junling; Zhou, Qiong

    2013-01-01

    Error-prone PCR was used to create more active or enantioselective variants of Penicillium expansum lipase (PEL). A variant with a valine to glycine substitution at residue 72 in the lid structure exhibited higher activity and enantioselectivity than those of wild-type PEL. Site-directed saturation mutagenesis was used to explore the sequence-function relationship and the substitution of Val72 of P. expansum lipase changed both catalytic activity and enantioselectivity greatly. The variant V72A, displayed a highest enantioselectivity enhanced to about twofold for the resolution of (R, S)-naproxen (E value increased from 104 to 200.7 for wild-type PEL and V72A variant, respectively). In comparison to PEL, the variant V72A showed a remarkable increase in specific activity towards p-nitrophenyl palmitate (11- and 4-fold increase at 25 and 35 °C, respectively) whereas it had a decreased thermostability. The results suggest that the enantioselective variant V72A could be used for the production of pharmaceutical drugs such as enantiomerically pure (S)-naproxen and the residue Val 72 of P. expansum lipase plays a significant role in the enantioselectivity and activity of this enantioselective lipase.

  12. Blue cheese-making has shaped the population genetic structure of the mould Penicillium roqueforti

    PubMed Central

    Ropars, Jeanne; López-Villavicencio, Manuela; Snirc, Alodie; Lacoste, Sandrine; Giraud, Tatiana

    2017-01-01

    Background Penicillium roqueforti is a filamentous fungus used for making blue cheeses worldwide. It also occurs as a food spoiler and in silage and wood. Previous studies have revealed a strong population genetic structure, with specific traits associated with the different populations. Here, we used a large strain collection from worldwide cheeses published recently to investigate the genetic structure of P. roqueforti. Principal findings We found a genetic population structure in P. roqueforti that was consistent with previous studies, with two main genetic clusters (W+C+ and W-C-, i.e., with and without horizontal gene transferred regions CheesyTer and Wallaby). In addition, we detected a finer genetic subdivision that corresponded to the environment and to protected designation of origin (PDO), namely the Roquefort PDO. We indeed found evidence for eight genetic clusters, one of the cluster including only strains from other environments than cheeses, and another cluster encompassing only strains from the Roquefort PDO. The W-C- and W+C+ cheese clusters were not the most closely related ones, suggesting that there may have been two independent domestication events of P. roqueforti for making blue cheeses. Significance The additional population structure revealed here may be relevant for cheese-makers and for understanding the history of domestication in P. roqueforti. PMID:28248964

  13. An Antifungal Role of Hydrogen Sulfide on the Postharvest Pathogens Aspergillus niger and Penicillium italicum

    PubMed Central

    Li, Yan-Hong; Hu, Liang-Bin; Yan, Hong; Liu, Yong-Sheng; Zhang, Hua

    2014-01-01

    In this research, the antifungal role of hydrogen sulfide (H2S) on the postharvest pathogens Aspergillus niger and Penicillium italicum growing on fruits and under culture conditions on defined media was investigated. Our results show that H2S, released by sodium hydrosulfide (NaHS) effectively reduced the postharvest decay of fruits induced by A. niger and P. italicum. Furthermore, H2S inhibited spore germination, germ tube elongation, mycelial growth, and produced abnormal mycelial contractions when the fungi were grown on defined media in Petri plates. Further studies showed that H2S could cause an increase in intracellular reactive oxygen species (ROS) in A. niger. In accordance with this observation we show that enzyme activities and the expression of superoxide dismutase (SOD) and catalase (CAT) genes in A. niger treated with H2S were lower than those in control. Moreover, H2S also significantly inhibited the growth of Saccharomyces cerevisiae, Rhizopus oryzae, the human pathogen Candida albicans, and several food-borne bacteria. We also found that short time exposure of H2S showed a microbicidal role rather than just inhibiting the growth of microbes. Taken together, this study suggests the potential value of H2S in reducing postharvest loss and food spoilage caused by microbe propagation. PMID:25101960

  14. Proteome Changes in Penicillium expansum Grown in a Medium Derived from Host Plant.

    PubMed

    Xia, Xiaoshuang; Li, Huan; Liu, Fei; Zhang, Ye; Zhang, Qi; Wang, Yun; Li, Peiwu

    2017-03-28

    Penicillium expansum causes blue mold rot, a prevalent postharvest disease of pome fruit, and is also the main producer of the patulin. However, knowledge on the molecular mechanisms involved in this pathogen-host interaction remains largely unknown. In this work, a two-dimensional gel electrophoresis-based proteomic approach was applied to probe changes in P. expansum 3.3703 cultivated in apple juice medium, which was used to mimic the in planta condition. The results showed that the pH value and reducing sugar content in the apple juice medium decreased whereas the patulin content increased with the growing of P. expansum. A total of 28 protein spots that were up-regulated in P. expansum when grown in apple juice medium were identified. Functional categorization revealed that the identified proteins were mainly related to carbohydrate metabolism, secondary metabolism, protein biosynthesis or degradation, and redox homeostasis. Remarkably, several induced proteins, including glucose dehydrogenase, galactose oxidase, and FAD-binding monooxygenase, which might be responsible for the observed medium acidification and patulin production, were also detected. Overall, the experimental results provide a comprehensive interpretation of the physiological and proteomic responses of P. expansum to the host plant environment, and future functional characterization of the identified proteins will deepen our understanding of fungi-host interactions.

  15. Paths and determinants for Penicillium janthinellum to resist low and high copper

    PubMed Central

    Xu, Jian; Chen, Guo-Li; Sun, Xue-Zhe; Fan, Xian-Wei; You-Zhi, Li

    2015-01-01

    Copper (Cu) tolerance was well understood in fungi yeasts but not in filamentous fungi. Filamentous fungi are eukaryotes but unlike eukaryotic fungi yeasts, which are a collection of various fungi that are maybe classified into different taxa but all characterized by growth as filamentous hyphae cells and with a complex morphology. The current knowledge of Cu resistance of filamentous fungi is still fragmental and therefore needs to be bridged. In this study, we characterized Cu resistance of Penicillium janthinellum strain GXCR and its Cu-resistance-decreasing mutants (EC-6 and UC-8), and conducted sequencing of a total of 6 transcriptomes from wild-type GXCR and mutant EC-6 grown under control and external Cu. Taken all the results together, Cu effects on the basal metabolism were directed to solute transport by two superfamilies of solute carrier and major facilitator, the buffering free CoA and Acyl-CoA pool in the peroxisome, F-type H+-transporting ATPases-based ATP production, V-type H+-transporting ATPases-based transmembrane transport, protein degradation, and alternative splicing of pre-mRNAs. Roles of enzymatic and non-enzymatic antioxidants in resistance to low and high Cu were defined. The backbone paths, signaling systems, and determinants that involve resistance of filamentous fungi to high Cu were determined, discussed and outlined in a model. PMID:26265593

  16. Optimization of Antioxidant Potential of Penicillium granulatum Bainier by Statistical Approaches

    PubMed Central

    Chandra, Priyanka; Arora, Daljit Singh

    2012-01-01

    A three-step optimization strategy which includes one-factor-at-a-time classical method and different statistical approaches (Plackett-Burman design and response surface methodology) that were applied to optimize the antioxidant potential of Penicillium granulatum. Antioxidant activity was assayed by different procedures and compared with total phenolic content. Primarily, different carbon and nitrogen sources were screened by classical methods, which revealed sucrose and NaNO3 to be the most suitable. In second step, Plackett-Burman design also supported sucrose and NaNO3 to be the most significant. In third step, response surface analysis showed 4.5% sucrose, 0.1% NaNO3, and incubation temperature of 25°C to be the optimal conditions. Under these conditions, the antioxidant potential assayed through different procedures was 78.2%, 70.1%, and 78.9% scavenging effect for DPPH radical, ferrous ion, and nitric oxide ion, respectively. The reducing power showed an absorbance of 1.6 with 68.5% activity for FRAP assay. PMID:23724323

  17. Detergent-like stressor and nutrient in metabolism of Penicillium chrysogenum.

    PubMed

    Jakovljević, Violeta; Milićević, Jasmina; Stojanović, Jelica

    2014-01-02

    The influence of detergents on the metabolism of Penicillium chrysogenum from two aspects, as a stress factor and potential nutrient, was studied. The fungus was isolated from the river bed Lepenica, Kragujevac, at a place where sewage domestic wastewater discharged into the river. The fungus was grown in a liquid nutrient medium according to Czapek with and without addition of commercial detergent (MERIX, Henkel, Serbia) at a concentration of 0.3% and 0.5%. The biochemical changes of pH, redox potential, free and total organic acids, total dry weight biomass, activity of alkaline and acid invertase and alkaline phosphatase were evaluated from day 3 to day 16 of the fungus growth. At the same time, detergent disappearance in terms of methylene blue active substances in the medium was measured. The detergent at a concentration of 0.5% showed a fungicide effect. In the medium with 0.3% of detergent, there was increased pH and concentration of organic acids, but decreased redox potential and total dry weight biomass. The detergent also showed an inhibitory effect on invertase and phosphatase activity. P. chrysogenum decomposed 50.2% of the total detergent concentration for an experimental period of 16 days.

  18. Kinetic and stability properties of Penicillium chrysogenum ATP sulfurylase missing the C-terminal regulatory domain.

    PubMed

    Hanna, Eissa; Ng, Kit Fai; MacRae, Ian J; Bley, Christopher J; Fisher, Andrew J; Segel, Irwin H

    2004-02-06

    ATP sulfurylase from Penicillium chrysogenum is a homohexameric enzyme that is subject to allosteric inhibition by 3'-phosphoadenosine 5'-phosphosulfate. In contrast to the wild type enzyme, recombinant ATP sulfurylase lacking the C-terminal allosteric domain was monomeric and noncooperative. All kcat values were decreased (the adenosine 5'-phosphosulfate (adenylylsulfate) (APS) synthesis reaction to 17% of the wild type value). Additionally, the Michaelis constants for MgATP and sulfate (or molybdate), the dissociation constant of E.APS, and the monovalent oxyanion dissociation constants of dead end E.MgATP.oxyanion complexes were all increased. APS release (the k6 step) was rate-limiting in the wild type enzyme. Without the C-terminal domain, the composite k5 step (isomerization of the central complex and MgPPi release) became rate-limiting. The cumulative results indicate that besides (a) serving as a receptor for the allosteric inhibitor, the C-terminal domain (b) stabilizes the hexameric structure and indirectly, individual subunits. Additionally, (c) the domain interacts with and perfects the catalytic site such that one or more steps following the formation of the binary E.MgATP and E.SO4(2-) complexes and preceding the release of MgPPi are optimized. The more negative entropy of activation of the truncated enzyme for APS synthesis is consistent with a role of the C-terminal domain in promoting the effective orientation of MgATP and sulfate at the active site.

  19. The nagA gene of Penicillium chrysogenum encoding beta-N-acetylglucosaminidase.

    PubMed

    Díez, Bruno; Rodríguez-Sáiz, Marta; de la Fuente, Juan Luis; Moreno, Miguel Angel; Barredo, José Luis

    2005-01-15

    We purified the beta-N-acetylglucosaminidase from the filamentous fungus Penicillium chrysogenum and its N-terminal sequence was determined, showing the presence of a mixture of two proteins (P1 and P2). A genomic DNA fragment was cloned by using degenerated oligonucleotides from the Nt sequences. The nucleotide sequence showed the presence of an ORF (nagA gene) lacking introns, with a length of 1791 bp, and coding for a protein of 66.5 kDa showing similarity to acetylglucosaminidases. The NagA deduced protein includes P1 and P2 as incomplete forms of the mature protein, and contains putative features for protein maturation: an 18-amino acid signal peptide, a KEX2 processing site, and four glycosylation motifs. The sequence just after the signal peptide corresponds to P2 and that after the KEX2 site to P1. The nagA transcript has a size of about 2.1 kb and is present until the end of the fermentation process for penicillin production. NagA is one of the most largely represented proteins in P. chrysogenum, increasing along the fermentation process. The suitability of the nagA promoter (PnagA) for gene expression in fungi was demonstrated by expressing the bleomycin resistance gene (ble(R)) from Streptoalloteichus hindustanus in P. chrysogenum.

  20. Biodegradation of triphenylmethane dye cotton blue by Penicillium ochrochloron MTCC 517.

    PubMed

    Shedbalkar, Utkarsha; Dhanve, Rhishikesh; Jadhav, Jyoti

    2008-09-15

    Triphenylmethane dyes belong to the most important group of synthetic colorants and are used extensively in the textile industries for dying cotton, wool, silk, nylon, etc. They are generally considered as the xenobiotic compounds, which are very recalcitrant to biodegradation. Penicillium ochrochloron decolorizes cotton blue (50 mg l(-1)) within 2.5 h under static condition at pH 6.5 and temperature 25 degrees C. TLC, FTIR and HPLC analysis confirms biodegradation of cotton blue. FTIR spectroscopy and GC-MS analysis indicated sulphonamide and triphenylmethane as the final products of cotton blue degradation. The pH, temperature and maturity of biomass affected the rate of decolorization. Presence of lignin peroxidase, tyrosinase and aminopyrine N-demethylase activities in the cell homogenate as well as increase in the extracellular activity of lignin peroxidase suggests the role of these enzymes in the decolorization process. The phytotoxicity and microbial toxicity studies of extracted metabolites suggest the less toxic nature of them.

  1. A microscopy study of hyphal growth of Penicillium rubens on gypsum under dynamic humidity conditions.

    PubMed

    van Laarhoven, Karel A; Huinink, Hendrik P; Adan, Olaf C G

    2016-05-01

    To remediate indoor fungal growth, understanding the moisture relations of common indoor fungi is crucial. Indoor moisture conditions are commonly quantified by the relative humidity (RH). RH is a major determinant of the availability of water in porous indoor surfaces that fungi grow on. The influence of steady-state RH on growth is well understood. Typically, however, the indoor RH constantly changes so that fungi have to endure frequent periods of alternating low and high RH. Knowledge of how common indoor fungi survive and are affected by the low-RH periods is limited. In particular, the specific effects of a drop in RH on the growth of the mycelium remain unclear. In this work, video microscopy was used to monitor hyphal growth of Penicillium rubens on gypsum substrates under controlled dynamic humidity conditions. The effect of a single period of low RH (RH = 50-90%) interrupting favourable conditions (RH = 97%) was tested. It was found that hyphal tips ceased to extend when exposed to any tested decrease in RH. However, new hyphal growth always emerges, seemingly from the old mycelium, suggesting that this indoor fungus does not rely only on conidia to survive the humidity patterns considered. These findings are a fundamental step in unravelling the effect of RH on indoor fungal growth.

  2. Role of Penicillium chrysogenum XJ-1 in the Detoxification and Bioremediation of Cadmium

    PubMed Central

    Xu, Xingjian; Xia, Lu; Zhu, Wei; Zhang, Zheyi; Huang, Qiaoyun; Chen, Wenli

    2015-01-01

    Microbial bioremediation is a promising technology to treat heavy metal-contaminated soils. However, the efficiency of filamentous fungi as bioremediation agents remains unknown, and the detoxification mechanism of heavy metals by filamentous fungi remains unclear. Therefore, in this study, we investigated the cell morphology and antioxidant systems of Penicillium chrysogenum XJ-1 in response to different cadmium (Cd) concentrations (0–10 mM) by using physico-chemical and biochemical methods. Cd in XJ-1 was mainly bound to the cell wall. The malondialdehyde level in XJ-1 cells was increased by 14.82–94.67 times with the increase in Cd concentration. The activities of superoxide dismutase, glutathione reductase (GR), and glucose-6-phosphate dehydrogenase (G6PDH) peaked at 1 mM Cd, whereas that of catalase peaked at 5 mM Cd. Cd exposure increased the glutathione/oxidized glutathione ratio and the activities of GR and G6PDH in XJ-1. These results suggested that the Cd detoxification mechanism of XJ-1 included biosorption, cellular sequestration, and antioxidant defense. The application of XJ-1 in Cd-polluted soils (5–50 mg kg-1) successfully reduced bioavailable Cd and increased the plant yield, indicating that this fungus was a promising candidate for in situ bioremediation of Cd-polluted soil. PMID:26733967

  3. Detection of Specific Antibodies to an Antigenic Mannoprotein for Diagnosis of Penicillium marneffei Penicilliosis

    PubMed Central

    Cao, Liang; Chen, Da-Liang; Lee, Cindy; Chan, Che-Man; Chan, King-Man; Vanittanakom, Nongnuch; Tsang, Dominic N. C.; Yuen, Kwok-Yung

    1998-01-01

    The disseminated and progressive fungal disease Penicillium marneffei penicilliosis is one of the most common infectious diseases in AIDS patients in Southeast Asia. To diagnose systemic penicilliosis, we developed an enzyme-linked immunosorbent assay (ELISA)-based antibody test with Mp1p, a purified recombinant antigenic mannoprotein of P. marneffei. Evaluation of the test with guinea pig sera against P. marneffei and other pathogenic fungi indicated that this assay was specific for P. marneffei. Clinical evaluation revealed that high levels of specific antibody were detected in two immunocompetent penicilliosis patients. Furthermore, approximately 80% (14 of 17) of the documented penicilliosis patients with human immunodeficiency virus tested positive for the specific antibody. No false-positive results were found for serum samples from 90 healthy blood donors, 20 patients with typhoid fever, and 55 patients with tuberculosis, indicating a high specificity of the test. Thus, this ELISA-based test for the detection of anti-Mp1p antibody can be of significant value as a diagnostic for penicilliosis. PMID:9738061

  4. The Mutation Breeding and Mutagenic Effect of Air Plasma on Penicillium Chrysogenum

    NASA Astrophysics Data System (ADS)

    Gui, Fang; Wang, Hui; Wang, Peng; Liu, Hui; Cai, Xiaochun; Hu, Yihua; Yuan, Chengling; Zheng, Zhiming

    2012-04-01

    Low temperature air plasma was used as the mutation tool for penicillin-producing strain Penicillium chrysogenum. The discharge conditions were RF power of 360 W, temperature of 40°C in a sealed chamber, and pressure of 10 Pa to 30 Pa. The result showed that the kinetics of the survival rate followed a typical saddle-shaped curve. Based on a statistic analysis, at the treating duration of 10 min, the positive mutation rate was as high as 37.5% while the negative mutation rate was low. The colonial morphology changed obviously when the plasma treating duration reached or exceeded 45 min. After both primary and secondary screening, a mutant designated as aPc051310 with high productivity of penicillin was obtained, and a strong mutagenic effect on P. chrysogenum was observed in the process. It was proved that after five generations, the mutant aPc051310 still exhibits a high productivity. All the results prove that the plasma mutation method could be developed as a convenient and effective tool to breed high-yield strains in the fermentation industry, while expanding the plasm application at the same time.

  5. Proteolytic activity of Penicillium chrysogenum and Debaryomyces hansenii during controlled ripening of pork loins.

    PubMed

    Martín, Alberto; Asensio, Miguel A; Bermúdez, María E; Córdoba, María G; Aranda, Emilio; Córdoba, Juan J

    2002-09-01

    The role of micro-organisms on the ripening process of dry-cured ham, particularly with respect to proteolysis, is not clear. This is partially due to the lack of an adequate system to study changes on a sterile control meat product for long ripening times. Using a meat system based on sterile pork loins ripened under aseptic conditions for 106 days, the contribution to the proteolysis of two micro-organisms isolated from dry-cured ham has been established. Changes were studied by SDS-PAGE of sarcoplasmic and myofibrillar proteins, capillary zone electrophoresis (CZE) of low ionic strength-soluble nitrogen compounds, and HPLC of free amino acids. Debaryomyces hansenii Dh345 did not show any significant proteolytic activity. However, Penicillium chrysogenum Pg222 showed high proteolytic activity on myofibrillar proteins resulting in an increase in soluble nitrogen compounds. For this, P. chrysogenum Pg222 should be considered to be used as starter culture in meat products made using long ripening times.

  6. Efficacy of salicylic acid to reduce Penicillium expansum inoculum and preserve apple fruits.

    PubMed

    da Rocha Neto, Argus Cezar; Luiz, Caroline; Maraschin, Marcelo; Di Piero, Robson Marcelo

    2016-03-16

    Apples are among the most commonly consumed fruits worldwide. Blue mold (Penicillium expansum) is one of the major diseases in apples postharvest, leading to wide use of fungicides and the search for alternative products to control the pathogen. In this context, this study aimed to evaluate the potential of salicylic acid (SA) as an alternative product to control blue mold and to preserve the physicochemical characteristics of apple fruit postharvest. The antimicrobial effect of SA was determined both in vitro and in situ, by directly exposing conidia to solutions of different concentrations SA or by inoculating the fruit with P. expansum and treating them curatively, eradicatively, or preventively with a 2.5mM SA solution. The physiological effects of SA on fruit were determined by quantifying the weight loss, total soluble solids content, and titratable acidity. In addition, the accumulation of SA in the fruit was determined by HPLC. SA (2.5mM) inhibited 100% of fungal germination in vitro and also controlled blue mold in situ when applied eradicatively. In addition, HPLC analysis demonstrated that SA did not persist in apple fruit. SA also maintained the physicochemical characteristics of fruit of different quality categories. Thus, SA may be an alternative to the commercial fungicides currently used against P. expansum.

  7. Antifungal activity of salicylic acid against Penicillium expansum and its possible mechanisms of action.

    PubMed

    da Rocha Neto, Argus Cezar; Maraschin, Marcelo; Di Piero, Robson Marcelo

    2015-12-23

    Apple is a fruit widely produced and consumed around the world. Blue mold (Penicillium expansum) is one of the main postharvest diseases in apples, leading to a wide use of fungicides and the search for alternative products. The aim of this study was to assess the effect of salicylic acid (SA) against P. expansum, elucidating its mechanisms of action. The antimicrobial effect was determined by exposing conidia to a 2.5 mM SA solution for 0 to 120 min, followed by incubation. The effect of pH on the efficacy of SA against P. expansum was assessed both in vitro and in situ. The action mechanisms were investigated through fluorescence assays, measurement of protein leakage, lipid damage, and transmission electronic microscopy. SA was capable of inhibiting 90% of the fungal germination after 30 min, causing damage to the conidial plasma membrane and leading to protein leakage up to 3.2 μg of soluble protein per g of mycelium. The pH of the SA solution affected the antimicrobial activity of this secondary metabolite, which inhibited the germination of P. expansum and the blue mold incidence in apples in solutions with pH≤3 by 100%, gradually losing its activity at higher pH.

  8. Effect of the incubation conditions on the production of patulin by Penicillium griseofulvum isolated from wheat.

    PubMed

    Jiménez, M; Mateo, R; Mateo, J J; Huerta, T; Hernández, E

    1991-09-01

    Sixty-four wheat samples from Spanish flour factories were screened for patulin and patulin-producing moulds. None of them was found to contain any patulin, whereas samples experimentally contaminated with this toxin proved it to be highly unstable. On the other hand, Penicillium griseofulvum was the only in vitro patulin-producing species found (19 samples). Mould growth in the samples was investigated by using yeast-sucrose medium (YES) and high-performance liquid chromatography (HPLC) to measure the amounts of toxin produced during 40 day's incubation at 20 and 28 degrees C. The highest yield rate of patulin was obtained between the 20th and 30th day of incubation; such a rate, however, was very low throughout the vigorous growth phase, during the first 20 days of incubation. The more appropriate temperature for incubation and patulin production was 28 degrees C. We also investigated the influence of other incubation conditions in the yield and found stationary dark cultures to be more efficient that shaken or fermentation cultures in YES medium. The best patulin yield achieved was 11.9 mg in the culture broth and 6.3 mg in the mycelium from 100 ml of medium.

  9. Fungal endophyte Penicillium janthinellum LK5 improves growth of ABA-deficient tomato under salinity.

    PubMed

    Khan, Abdul Latif; Waqas, Muhammad; Khan, Abdur Rahim; Hussain, Javid; Kang, Sang-Mo; Gilani, Syed Abdullah; Hamayun, Muhammad; Shin, Jae-Ho; Kamran, Muhammad; Al-Harrasi, Ahmed; Yun, Byung-Wook; Adnan, Muhammad; Lee, In-Jung

    2013-11-01

    An endophytic fungus was isolated from the roots of tomato (Solanum lycopersicum Mill) and identified as Penicillium janthinellum LK5. The culture filtrate (CF) of P. janthinellum significantly increased the shoot length of gibberellins (GAs) deficient mutant waito-c and normal Dongjin-beyo rice seedlings as compared to control. The CF of P. janthinellum contained GAs (GA3, GA4, GA7 and GA12). To assess endophyte-growth promoting and stress-tolerance potential, the CF along with the propagules of endophyte was applied to tomato-host and abscisic acid (ABA)-deficient mutant Sitiens plants under sodium chloride (NaCl) induced salinity stress. Sitiens plants had retarded growth under normal and salinity stress however its growth was much improved during P. janthinellum-association. The endophyte inoculation reduced the membrane injury by decreasing lipid peroxidation as compared to non-inoculated control under salinity. Endophyte-associated Sitiens plants have significantly higher catalase, peroxidase and glutathione activities as compared to control. Endophyte-infected host and Sitiens plants had low level of sodium ion toxicity and high calcium contents in its root as compared to control. P. janthinellum LK5 helped the Sitiens plants to synthesis significantly higher ABA and reduced the level of jasmonic acid to modulate stress responses. The results suggest that endophytes-association can resist salinity stress by producing gibberellins and activating defensive mechanisms of host and Sitiens plants to achieve improved growth.

  10. Production, Purification, and Characterization of a Major Penicillium glabrum Xylanase Using Brewer's Spent Grain as Substrate

    PubMed Central

    Beitel, Susan Michelz; Fortkamp, Diana; Terrasan, César Rafael Fanchini; de Almeida, Alex Fernando

    2013-01-01

    In recent decades, xylanases have been used in many processing industries. This study describes the xylanase production by Penicillium glabrum using brewer's spent grain as substrate. Additionally, this is the first work that reports the purification and characterization of a xylanase using this agroindustrial waste. Optimal production was obtained when P. glabrum was grown in liquid medium in pH 5.5, at 25 °C, under stationary condition for six days. The xylanase from P. glabrum was purified to homogeneity by a rapid and inexpensive procedure, using ammonium sulfate fractionation and molecular exclusion chromatography. SDS-PAGE analysis revealed one band with estimated molecular mass of 18.36 kDa. The optimum activity was observed at 60 °C, in pH 3.0. The enzyme was very stable at 50 °C, and high pH stability was verified from pH 2.5 to 5.0. The ion Mn2+ and the reducing agents β-mercaptoethanol and DTT enhanced xylanase activity, while the ions Hg2+, Zn2+, and Cu2+ as well as the detergent SDS were strong inhibitors of the enzyme. The use of brewer's spent grain as substrate for xylanase production cannot only add value and decrease the amount of this waste but also reduce the xylanase production cost. PMID:23762855

  11. Production, purification, and characterization of a major Penicillium glabrum xylanase using Brewer's spent grain as substrate.

    PubMed

    Knob, Adriana; Beitel, Susan Michelz; Fortkamp, Diana; Terrasan, César Rafael Fanchini; de Almeida, Alex Fernando

    2013-01-01

    In recent decades, xylanases have been used in many processing industries. This study describes the xylanase production by Penicillium glabrum using brewer's spent grain as substrate. Additionally, this is the first work that reports the purification and characterization of a xylanase using this agroindustrial waste. Optimal production was obtained when P. glabrum was grown in liquid medium in pH 5.5, at 25 °C, under stationary condition for six days. The xylanase from P. glabrum was purified to homogeneity by a rapid and inexpensive procedure, using ammonium sulfate fractionation and molecular exclusion chromatography. SDS-PAGE analysis revealed one band with estimated molecular mass of 18.36 kDa. The optimum activity was observed at 60 °C, in pH 3.0. The enzyme was very stable at 50 °C, and high pH stability was verified from pH 2.5 to 5.0. The ion Mn(2+) and the reducing agents β -mercaptoethanol and DTT enhanced xylanase activity, while the ions Hg(2+), Zn(2+), and Cu(2+) as well as the detergent SDS were strong inhibitors of the enzyme. The use of brewer's spent grain as substrate for xylanase production cannot only add value and decrease the amount of this waste but also reduce the xylanase production cost.

  12. Function of a p24 Heterodimer in Morphogenesis and Protein Transport in Penicillium oxalicum

    PubMed Central

    Wang, Fangzhong; Liu, Kuimei; Han, Lijuan; Jiang, Baojie; Wang, Mingyu; Fang, Xu

    2015-01-01

    The lignocellulose degradation capacity of filamentous fungi has been widely studied because of their cellulase hypersecretion. The p24 proteins in eukaryotes serve important functions in this secretory pathway. However, little is known about the functions of the p24 proteins in filamentous fungi. In this study, four p24 proteins were identified in Penicillium oxalicum. Six p24 double-deletion strains were constructed, and further studies were carried out with the ΔerpΔpδ strain. The experimental results suggested that Erp and Pδ form a p24 heterodimer in vivo. This p24 heterodimer participates in important morphogenetic events, including sporulation, hyphal growth, and lateral branching. The results suggested that the p24 heterodimer mediates protein transport, particularly that of cellobiohydrolase. Analysis of the intracellular proteome revealed that the ΔerpΔpδ double mutant is under secretion stress due to attempts to remove proteins that are jammed in the endomembrane system. These results suggest that the p24 heterodimer participates in morphogenesis and protein transport. Compared with P. oxalicum Δerp, a greater number of cellular physiological pathways were impaired in ΔerpΔpδ. This finding may provide new insights into the secretory pathways of filamentous fungi. PMID:26149342

  13. Enhanced cellulase production by Penicillium oxalicum for bio-ethanol application.

    PubMed

    Saini, Reetu; Saini, Jitendra Kumar; Adsul, Mukund; Patel, Anil Kumar; Mathur, Anshu; Tuli, Deepak; Singhania, Reeta Rani

    2015-01-01

    Present study was focused on cellulase production from an indigenously isolated filamentous fungal strain, identified as Penicillium oxalicum. Initially, cellulase production under submerged fermentation in shake flasks resulted in cellulase activity of 0.7 FPU/mL. Optimization of process parameters enhanced cellulase production by 1.7-fold and resulted in maximum cellulase activity of 1.2 FPU/mL in 8 days. Cellulase production was successfully scaled-up to 7 L fermenter under controlled conditions and incubation time was reduced from 8 days to 4 days for achieving similar cellulase titer. Optimum pH and temperature for activity of the crude enzyme were pH 5 and 50 °C, respectively. At 50 °C the produced cellulase retained approximately 50% and 26% of its activity at 48 h and 72 h, respectively. Hydrolytic efficiency of P. oxalicum was comparable to commercial cellulase preparations which indicate its great potential for application in the lignocellulose hydrolysis.

  14. A Newly Isolated Penicillium oxalicum 16 Cellulase with High Efficient Synergism and High Tolerance of Monosaccharide.

    PubMed

    Zhao, Xi-Hua; Wang, Wei; Tong, Bin; Zhang, Su-Ping; Wei, Dong-Zhi

    2016-01-01

    Compared to Trichoderma reesei RUT-C30 cellulase (Trcel), Penicillium oxalicum 16 cellulase (P16cel) from the fermentation supernatant produced a 2-fold higher glucose yield when degrading microcrystalline cellulose (MCC), possessed a 10-fold higher β-glucosidase (BGL) activity, but obtained somewhat lower other cellulase component activities. The optimal temperature and pH of β-1,4-endoglucanase, cellobiohydrolase, and filter paperase from P16cel were 50-60 °C and 4-5, respectively, but those of BGL reached 70 °C and 5. The cellulase cocktail of P16cel and Trcel had a high synergism when solubilizing MCC and generated 1.7-fold and 6.2-fold higher glucose yields than P16cel and Trcel at the same filter paperase loading, respectively. Additional low concentration of fructose enhanced the glucose yield during enzymatic hydrolysis of MCC; however, additional high concentration of monosaccharide (especially glucose) reduced cellulase activities and gave a stronger monosaccharide inhibition on Trcel. These results indicate that P16cel is a more excellent cellulase than Trcel.

  15. Alteration of Acrylonitrile-Methylacrylate-Butadiene Terpolymer by Nocardia rhodochrous and Penicillium notatum†

    PubMed Central

    Antoine, A. D.; Dean, A. V.; Gilbert, S. G.

    1980-01-01

    [14C]Barex-210, a terpolymer of acrylonitrile, methylacrylate, and butadiene, was tested for bioconversion. Powdered samples of polymer, each specifically 14C labeled at different carbon atoms of the polymer, were incubated with either Nocardia rhodochrous or Penicillium notatum in an enriched growth medium for various periods of time. After 6 months of incubation, the 14C-labeled polymer was transformed from a high-molecular-weight material completely soluble in dimethyl formamide (DMF) into both a lower-molecular-weight form still soluble in DMF and a second form that was no longer soluble in DMF. The amount of 14C-labeled carbon atoms converted into DMF-insoluble material was 8% of the backbone carbon-carbon atoms and 12% of the side-chain nitrile and acrylate atoms from the acrylonitrile-methylacrylate copolymer and 60% of the elastomer (acrylonitrile-butadiene copolymer) atoms. Metabolism of the polymer was not established from measurements of metabolic 14CO2. Evolution of 14CO2 amounted to only 0.3, 0.6, 1.8, and 3.3% of these four fractions, respectively. Although the transformation of high-molecular-weight polymer into DMF-insoluble material was rapid in the early stages of microbial growth, the accompanying CO2 evolution was much slower. Further evidence of polymer alteration was indicated by the infrared spectrum of the insoluble material, which showed a disappearance of the nitrile and methylacrylate peaks. PMID:16345541

  16. Improvement of submerged culture conditions to produce colorants by Penicillium purpurogenum.

    PubMed

    Santos-Ebinuma, Valéria Carvalho; Roberto, Inês Conceição; Teixeira, Maria Francisca Simas; Pessoa, Adalberto

    2014-01-01

    Safety issues related to the employment of synthetic colorants in different industrial segments have increased the interest in the production of colorants from natural sources, such as microorganisms. Improved cultivation technologies have allowed the use of microorganisms as an alternative source of natural colorants. The objective of this work was to evaluate the influence of some factors on natural colorants production by a recently isolated from Amazon Forest, Penicillium purpurogenum DPUA 1275 employing statistical tools. To this purpose the following variables: orbital stirring speed, pH, temperature, sucrose and yeast extract concentrations and incubation time were studied through two fractional factorial, one full factorial and a central composite factorial designs. The regression analysis pointed out that sucrose and yeast extract concentrations were the variables that influenced more in colorants production. Under the best conditions (yeast extract concentration around 10 g/L and sucrose concentration of 50 g/L) an increase of 10, 33 and 23% respectively to yellow, orange and red colorants absorbance was achieved. These results show that P. purpurogenum is an alternative colorants producer and the production of these biocompounds can be improved employing statistical tool.

  17. Gene encoding inulinase isolated from Penicillium citrinum ESS and its molecular phylogeny.

    PubMed

    Flores-Gallegos, Adriana C; Morlett-Chávez, Jesús A; Aguilar, Cristóbal N; Riutort, Marta; Rodríguez-Herrera, Raúl

    2015-02-01

    Inulinase is an enzyme produced by plants and several microorganisms, including fungi, to hydrolyze the β-2,1 glycosidic linkages present in some oligosaccharides to produce fructose and glucose. This enzyme, in conjunction with invertases, levanases, and two types of 1-fructosyl transferases have been described as members of the glycosyl hydrolases (family 32), the most diverse group of enzymes used by microbes for biomass degradation. As being part of the same clan, they have common evolutionary origin sharing the most important functional characteristics. Recently, a xerophylic fungi strain isolated from Mexican semi-desert, Penicillium citrinum ESS has been reported as inulinase producer, which could have greater stability than other enzymes due to a metabolic machinery adapted to typical temperature changes in this region. To continue the understanding of action mechanisms of these enzymes and to establish evolutionary relationships within this family, in the present study, phylogenetic analyses were used to analyze amino acid sequences coding fungal and yeast glycoside hydrolases of family 32, including the new sequenced inulinase of P. citrinum ESS. It was possible to elucidate the action mechanism of fungal glycoside hydrolases in present study and to classify inulinase from P. citrinum ESS as an exo-inulinase on the basis of their amino acid sequence phylogenetic affinities.

  18. Increased Penicillin Production in Penicillium chrysogenum Production Strains via Balanced Overexpression of Isopenicillin N Acyltransferase

    PubMed Central

    Weber, Stefan S.; Polli, Fabiola; Boer, Rémon; Bovenberg, Roel A. L.

    2012-01-01

    Intense classical strain improvement has yielded industrial Penicillium chrysogenum strains that produce high titers of penicillin. These strains contain multiple copies of the penicillin biosynthesis cluster encoding the three key enzymes: δ-(l-α-aminoadipyl)-l-cysteinyl-d-valine synthetase (ACVS), isopenicillin N synthase (IPNS), and isopenicillin N acyltransferase (IAT). The phenylacetic acid coenzyme A (CoA) ligase (PCL) gene encoding the enzyme responsible for the activation of the side chain precursor phenylacetic acid is localized elsewhere in the genome in a single copy. Since the protein level of IAT already saturates at low cluster copy numbers, IAT might catalyze a limiting step in high-yielding strains. Here, we show that penicillin production in high-yielding strains can be further improved by the overexpression of IAT while at very high levels of IAT the precursor 6-aminopenicillic acid (6-APA) accumulates. Overproduction of PCL only marginally stimulates penicillin production. These data demonstrate that in high-yielding strains IAT is the limiting factor and that this limitation can be alleviated by a balanced overproduction of this enzyme. PMID:22865068

  19. Bioleaching of tungsten-rich spent hydrocracking catalyst using Penicillium simplicissimum.

    PubMed

    Amiri, F; Yaghmaei, S; Mousavi, S M

    2011-01-01

    Adaptation of Penicillium simplicissimum with different heavy metals present in a spent hydrocracking catalyst, as well as one-step, two-step, and spent medium bioleaching of the spent catalyst by the adapted fungus, was examined in batch cultures. Adaptation experiments with the single metal ions Ni, Mo, Fe, and W showed that the fungus could tolerate up to 1500 mg/L Ni, 8000 mg/L Mo, 3000 mg/L Fe, and 8000 mg/L W. In the presence of multi-metals, the fungus was able to tolerate up to 300 mg/L Ni, 200 mg/L Mo, 150 mg/L Fe and 2500 mg/L W. A total of 3% (w/v) spent catalyst generally gave the maximum extraction yields in the two-step bioleaching process (100% of W, 100% of Fe, 92.7% of Mo, 66.43% of Ni, and 25% of Al). The main lixiviant in the bioleaching was shown to be gluconic acid. The red pigment produced by the fungus could also possibly act as an agent in Al leaching.

  20. Production of an Extracellular Matrix as an Isotropic Growth Phase of Penicillium rubens on Gypsum

    PubMed Central

    Bekker, M.; Adan, O. C. G.; Samson, R. A.; Wyatt, T.; Dijksterhuis, J.

    2012-01-01

    Indoor mold represents an important environmental concern, but a fundamental knowledge of fungal growth stages is needed to limit indoor fungal proliferation on finishing materials used in buildings. The present study focused on the succession of germination stages of the common indoor fungus Penicillium rubens on a gypsum substrate. This substrate is used as a model system representing porous materials that are widely used in indoor environments. Imaging with cryo-scanning electron microscopy showed that the formation of an extracellular matrix (ECM) is a phase of the isotropic growth of P. rubens that is uniquely related to germinating conidia. Furthermore, the ECM is observed only when a dry-state inoculation of the surface is applied, i.e., applying conidia directly from a 7-day-old colony, mimicking airborne contamination of the surface. When inoculation is done by spraying an aqueous conidial suspension, no ECM is observed. Moreover, it is concluded that the formation of an ECM requires active processes in the fungal cell. The porosity of the substrate proved that the ECM substance has high-viscosity characteristics. The present results stress that studies of indoor fungal growth should consider the method of inoculation, knowing that the common aqueous suspension may obscure specific stages in the initial phases of germination. PMID:22843536

  1. Penicillium chrysogenum DSOA associated with marine sponge (Tedania anhelans) exhibit antimycobacterial activity.

    PubMed

    Visamsetti, Amarendra; Ramachandran, Santhosh Sarojini; Kandasamy, Dhevendaran

    2016-04-01

    A strain of Penicillium chrysogenum was isolated from Tedania anhelans (marine sponge) collected from Indian Ocean (8°22'30″N latitude and 76°59'16″ longitude) and deposited in culture collection centers. The strain subjected to different culture conditions for production of extrolites were extracted using ethyl acetate and chloroform. When both extracts were subjected for antibacterial activity, latter had high activity. Minimum inhibitory concentration of chloroform extract ranged from 31.25-1000 μg/mL in tested microbes such as, Mycobacterium tuberculosis H37Ra, Mycobacterium avium, Mycobacterium fortuitum, Mycobacterium smegmatis, Mycobacterium vaccae, Staphylococcus aureus, Aeromonas hydrophila, Pseudomonas aeruginosa and Vibrio cholerae. No cytotoxicity was observed in Vero cell line up to 399.10 μg/mL. Antibacterial activity previously reported by Parameswaran et al. in 1997 from ethyl acetate extract of T. anhelans might be due to the diketopiperazines, Cyclo-(L-Pro-L-Phe) and Cyclo-(L-Leu-L-Pro) produced by the associated fungi-P. chrysogenum DSOA. It is producing a metabolites having antimycobacterial activity, a first report.

  2. Cellulases from Penicillium funiculosum: production, properties and application to cellulose hydrolysis.

    PubMed

    de Castro, Aline Machado; de Albuquerque de Carvalho, Marcelle Lins; Leite, Selma Gomes Ferreira; Pereira, Nei

    2010-02-01

    The objective of this work is to investigate the utilization of two abundant agricultural residues in Brazil for the production and application of cellulolytic enzymes. Different materials obtained after pretreatment of sugarcane bagasse, as well as pure synthetic substrates, were considered for cellulase production by Penicillium funiculosum. The best results for FPase (354 U L(-1)) and beta-glucosidase (1,835 U L(-1)) production were observed when sugarcane bagasse partially delignified cellulignin (PDC) was used. The crude extract obtained from PDC fermentation was then partially characterized. Optimal temperatures for cellulase action ranged from 52 to 58 degrees C and pH values of around 4.9 contributed to maximum enzyme activity. At 37 degrees C, the cellulases were highly stable, losing less than 15% of their initial activity after 23 h of incubation. There was no detection of proteases in the P. funiculosum extract, but other hydrolases, such as endoxylanases, were identified (147 U L(-1)). Finally, when compared to commercial preparations, the cellulolytic complex from P. funiculosum showed more well-balanced amounts of beta-glucosidase, endo- and exoglucanase, resulting in the desired performance in the presence of a lignocellulosic material. Cellulases from this filamentous fungus had a higher glucose production rate (470 mg L(-1) h(-1)) when incubated with corn cob than with Celluclast, GC 220 and Spezyme (312, 454 and 400 mg L(-1) h(-1), respectively).

  3. Alkaloids and polyketides from Penicillium citrinum, an endophyte isolated from the Moroccan plant Ceratonia siliqua.

    PubMed

    El-Neketi, Mona; Ebrahim, Weaam; Lin, Wenhan; Gedara, Sahar; Badria, Farid; Saad, Hassan-Elrady A; Lai, Daowan; Proksch, Peter

    2013-06-28

    The endophytic fungus Penicillium citrinum was isolated from a fresh stem of the Moroccan plant Ceratonia siliqua. Extracts of P. citrinum grown on rice and white bean media yielded five new compounds, namely, citriquinochroman (1), tanzawaic acids G and H (2 and 3), 6-methylcurvulinic acid (4), 8-methoxy-3,5-dimethylisoquinolin-6-ol (5), and one new natural product, 1,2,3,11b-tetrahydroquinolactacide (6), which had previously been described as a synthetic compound. In addition, 13 known compounds including seven alkaloids and six polyketides were isolated. The structures of the new compounds were unambiguously determined on the basis of one- and two-dimensional NMR spectroscopy as well as by high-resolution mass spectrometry. Citriquinochroman (1) features a new skeleton, consisting of quinolactacide and (3S)-6-hydroxy-8-methoxy-3,5-dimethylisochroman linked by a C-C bond. 1,2,3,11b-Tetrahydroquinolactacide (6) may be a biogenetic precursor of quinolactacide. Citriquinochroman (1) showed cytotoxicity against the murine lymphoma L5178Y cell line with an IC50 value of 6.1 μM, while the other compounds were inactive (IC50 >10 μM) in this assay.

  4. Properties of enzyme preparations and homogeneous enzymes - endoglucanases EG2 Penicillium verruculosum and LAM Myceliophthora thermophila.

    PubMed

    Merzlov, D A; Zorov, I N; Dotsenko, G S; Denisenko, Yu A; Rozhkova, A M; Satrutdinov, A D; Rubtsova, E A; Kondratieva, E G; Sinitsyn, A P

    2015-04-01

    The genes of endoglucanases EG2 (36.2 kDa) Penicillium verruculosum and LAM (30.8 kDa) Myceliophthora thermophila were cloned in P. verruculosum recombinant strain. New enzyme preparations with highly stable activity against β-glucan and laminarin were obtained and investigated, homogeneous enzymes EG2 (EC 3.2.1.4) and LAM (EC 3.2.1.6) being purified and characterized. For β-glucan, the EG2 Km value was found to be 10 times higher than that for LAM; however, EG2 demonstrated greater processivity due to its higher kcat. The pH and temperature optima of EG2 and LAM activity against barley β-glucan overlapped and were 4.3-4.9 and 61-67°C, respectively, and EG2 appeared to be more stable than LAM. Oligosaccharides with degree of polymerization 2-10 were formed by hydrolysis of β-glucan and laminarin by the studied enzymes. The recombinant enzyme preparations were faster and more effective in decreasing the reduced viscosity of wholegrain barley extract than some commercial enzyme preparations. Thus, the new enzyme preparations seem to be rather perspective as feed additives for degradation of non-starch polysaccharides in grain animal feed.

  5. Purification and characterization of an extracellular alkaline phosphatase from Penicillium chrysogenum.

    PubMed

    Politino, M; Brown, J; Usher, J J

    1996-01-01

    An extracellular alkaline phosphatase from Penicillium chrysogenum was purified to homogeneity using DEAE ion-exchange chromatography and size exclusion chromatography. SDS-PAGE of the purified enzyme indicated a molecular weight of 58,000. The mobility of the native enzyme on a Superose 12 column suggests that the active form of the enzyme is a monomer. The enzyme catalyzes the hydrolysis of phosphate from a variety of substrates including p-nitrophenyl phosphate, alpha-naphthyl phosphate and the anti-tumor compound etoposide phosphate. The apparent K(m) for the substrate p-nitrophenyl phosphate is 1.3 mM and the enzyme is inhibited by inorganic phosphate. The pH optimum of the enzyme is 9.0 with a broad optimal temperature range between 40 and 50 degrees C. The isoelectric point of the enzyme is approximately 5.5. The enzyme is a glycoprotein; digestion with endoglycosidase H indicates that the protein consists primarily of N-linked carbohydrates. Enzymatic activity is enhanced by the addition of divalent cations such as Mg+2 and Mn+2 and inhibited by addition of a chelator such as EDTA suggesting a metal ion requirement. The enzyme was found to be an inexpensive catalyst for the conversion of etoposide phosphate to etoposide in the manufacture of this anti-tumor compound.

  6. [Production of pectinases by Penicillium simplicissimum A3263 in an amaranth-seed flour medium].

    PubMed

    Pastor, M D; Lorda, G S; Balatti, A

    2002-01-01

    The present work studies the production of pectinases using a strain of Penicillium simplicissimum A3263 and considering the influence of adding Amaranthus cruentus seed meal in a selected medium. We also considered the influence of aeration on enzyme production. Research was oriented towards the production of pectin lyase, the enzyme having the highest commercial value. This work was carried out in Erlenmeyer flasks in rotary shaker to select the medium and in a mechanically stirred fermentor to study aeration. The microorganism was developed as pellets of 1 mm diameter. Enzyme levels were of the order of 8216.21 pectin lyase units and 167.57 polygalacturonase units per gram of fungal biomass, respectively, using a medium containing 40 g/l of amaranth seed meal. As for the influence of aeration, it was determined that the higher values were obtained at 750 rpm corresponding to an oxygen absorption rate of 2691 ml O2/lh for an air flow of 1 l/l.min. The results obtained are considered very important in view of the fact that they exceeded in 550% those obtained by other authors.

  7. Paths and determinants for Penicillium janthinellum to resist low and high copper.

    PubMed

    Xu, Jian; Chen, Guo-Li; Sun, Xue-Zhe; Fan, Xian-Wei; You-Zhi, Li

    2015-08-12

    Copper (Cu) tolerance was well understood in fungi yeasts but not in filamentous fungi. Filamentous fungi are eukaryotes but unlike eukaryotic fungi yeasts, which are a collection of various fungi that are maybe classified into different taxa but all characterized by growth as filamentous hyphae cells and with a complex morphology. The current knowledge of Cu resistance of filamentous fungi is still fragmental and therefore needs to be bridged. In this study, we characterized Cu resistance of Penicillium janthinellum strain GXCR and its Cu-resistance-decreasing mutants (EC-6 and UC-8), and conducted sequencing of a total of 6 transcriptomes from wild-type GXCR and mutant EC-6 grown under control and external Cu. Taken all the results together, Cu effects on the basal metabolism were directed to solute transport by two superfamilies of solute carrier and major facilitator, the buffering free CoA and Acyl-CoA pool in the peroxisome, F-type H(+)-transporting ATPases-based ATP production, V-type H(+)-transporting ATPases-based transmembrane transport, protein degradation, and alternative splicing of pre-mRNAs. Roles of enzymatic and non-enzymatic antioxidants in resistance to low and high Cu were defined. The backbone paths, signaling systems, and determinants that involve resistance of filamentous fungi to high Cu were determined, discussed and outlined in a model.

  8. Influence of storage temperature and apple variety on patulin production by Penicillium expansum.

    PubMed

    Salomão, Beatriz C M; Aragão, Gláucia M F; Churey, John J; Padilla-Zakour, Olga I; Worobo, Randy W

    2009-05-01

    This study examined the potential for patulin production in six different varieties of apples (Red Delicious, Golden Supreme, Gala, Fuji, Empire, and McIntosh) inoculated with Penicillium expansum spores and stored at two different temperatures (11 and 20.5 degrees C). Samples for patulin analysis were randomly taken from apples stored at different times, ranging from 21 to 93 days. While patulin was produced at both storage temperatures, apples incubated at 20.5 degrees C yielded significantly higher patulin concentrations than did those incubated at 11 degrees C. All apple varieties showed mold spoilage at both temperatures, except Red Delicious and Empire. A total of 44% of the samples analyzed showed patulin concentrations above the U.S. Food and Drug Administration regulatory limit (50 ppb). The highest patulin productions occurred in Golden Supreme (54,221 ppb) and McIntosh (52,131 and 48,457 ppb) varieties. Our results showed that careful culling of apples is essential for high juice quality, since high patulin levels in some apples varieties could result in a level greater than 50 ppb of this mycotoxin in the finished juice or cider, even when only one contaminated apple occurs in 1,000 apples.

  9. Strain-Specific Synthesis of Mycophenolic Acid by Penicillium roqueforti in Blue-Veined Cheese

    PubMed Central

    Engel, Günter; von Milczewski, Karl Ernst; Prokopek, Dieter; Teuber, Michael

    1982-01-01

    Twenty of 80 strains of Penicillium roqueforti were able to produce up to 600 mg of mycophenolic acid (MPA) liter−1 in 2% yeast extract-5% sucrose broth. Sixty-two of these strains had been isolated from the main blue-veined cheese varieties of western Europe or from starter cultures. Of these 62 dairy strains, only 7 had MPA-producing potential in vitro. These seven strains had all been isolated during the period 1975 to 1981 from the blue cheese of one individual factory. In cheese from the market, MPA (up to 5 mg kg−1) was only found in samples of this same factory. With MPA-producing and -nonproducing strains for the experimental manufacture of blue cheese, MPA synthesis in cheese was only detected with strains which form MPA in yeast extract-sucrose broth. The maximum MPA level at 4 mg kg−1 was similar to that in commercial cheese. Toxicity of MPA was tested with two established human cell lines (Detroit 98 and Girardi Heart) and one established pig kidney cell line (AmII). PMID:16346004

  10. Novel small molecule 11β-HSD1 inhibitor from the endophytic fungus Penicillium commune

    PubMed Central

    Sun, Weiguang; Chen, Xintao; Tong, Qingyi; Zhu, Hucheng; He, Yan; Lei, Liang; Xue, Yongbo; Yao, Guangmin; Luo, Zengwei; Wang, Jianping; Li, Hua; Zhang, Yonghui

    2016-01-01

    Two new phenone derivatives penicophenones A (1) and B (2), a new cyclic tetrapeptide penicopeptide A (3), and five known compounds were isolated from the culture broth of Penicillium commune, an endophytic fungus derived from Vitis vinifera. Compounds 1–3 were elucidated by extensive spectroscopic analyses including 1D and 2D NMR and HRESIMS. The absolute configurations of 1 and 3 were determined by comparing its ECD with related molecules and modified Marfey’s analysis, respectively. Penicophenone A (1) possesses a rare benzannulated 6,6-spiroketal moiety, which is a new member of the unusual structural class with peniphenone A as the representative. Compound 3 exhibited significant inhibition activities against 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) in vitro and showed strong binding affinity to 11β-HSD1. Moreover, compound 3 treatments decreased the lipid droplet accumulation associate with the inhibition of 11β-HSD1 expression in differentiate-induced 3T3-L1 preadipocytes. Furthermore, the molecular docking demonstrated that compound 3 coordinated in the active site of 11β-HSD1 is essential for the ability of diminishing the enzyme activity. PMID:27194583

  11. Fertility depression among cheese-making Penicillium roqueforti strains suggests degeneration during domestication.

    PubMed

    Ropars, Jeanne; Lo, Ying-Chu; Dumas, Emilie; Snirc, Alodie; Begerow, Dominik; Rollnik, Tanja; Lacoste, Sandrine; Dupont, Joëlle; Giraud, Tatiana; López-Villavicencio, Manuela

    2016-09-01

    Genetic differentiation occurs when gene flow is prevented, due to reproductive barriers or asexuality. Investigating the early barriers to gene flow is important for understanding the process of speciation. Here, we therefore investigated reproductive isolation between different genetic clusters of the fungus Penicillium roqueforti, used for maturing blue cheeses, and also occurring as food spoiler or in silage. We investigated premating and postmating fertility between and within three genetic clusters (two from cheese and one from other substrates), and we observed sexual structures under scanning electron microscopy. All intercluster types of crosses showed some fertility, suggesting that no intersterility has evolved between domesticated and wild populations despite adaptation to different environments and lack of gene flow. However, much lower fertility was found in crosses within the cheese clusters than within the noncheese cluster, suggesting reduced fertility of cheese strains, which may constitute a barrier to gene flow. Such degeneration may be due to bottlenecks during domestication and/or to the exclusive clonal replication of the strains in industry. This study shows that degeneration has occurred rapidly and independently in two lineages of a domesticated species. Altogether, these results inform on the processes and tempo of degeneration and speciation.

  12. Induction of sexual reproduction and genetic diversity in the cheese fungus Penicillium roqueforti.

    PubMed

    Ropars, Jeanne; López-Villavicencio, Manuela; Dupont, Joëlle; Snirc, Alodie; Gillot, Guillaume; Coton, Monika; Jany, Jean-Luc; Coton, Emmanuel; Giraud, Tatiana

    2014-04-01

    The emblematic fungus Penicillium roqueforti is used throughout the world as a starter culture in the production of blue-veined cheeses. Like other industrial filamentous fungi, P. roqueforti was thought to lack a sexual cycle. However, an ability to induce recombination is of great economic and fundamental importance, as it would make it possible to transform and improve industrial strains, promoting the creation of novel phenotypes and eliminating the deleterious mutations that accumulate during clonal propagation. We report here, for the first time, the induction of the sexual structures of P. roqueforti - ascogonia, cleistothecia and ascospores. The progeny of the sexual cycle displayed clear evidence of recombination. We also used the recently published genome sequence for this species to develop microsatellite markers for investigating the footprints of recombination and population structure in a large collection of isolates from around the world and from different environments. Indeed, P. roqueforti also occurs in silage, wood and human-related environments other than cheese. We found tremendous genetic diversity within P. roqueforti, even within cheese strains and identified six highly differentiated clusters that probably predate the use of this species for cheese production. Screening for phenotypic and metabolic differences between these populations could guide future development strategies.

  13. Ultrastructure, glutathione and low molecular weight proteins of Penicillium brevicompactum in response to cobalt.

    PubMed

    Farrag, Rasha M

    2009-01-01

    Penicillium brevicompactum highly tolerated cobalt concentrations of 50, 200, 800 and 1000 ppm both through cell wall and intracellular sequestration- immobilization of the metal on/within the cell wall, cell wall thickness, presence of electron-dense deposits inside vacuoles (thiol peptides sequestering cobalt) and in the cytoplasm (cobalt), and presence of matrixed electron-dense deposits, only at 800 and 1000 ppm, were observed. Increased vacuole formation and plasmolysis were also observed. Fraction number 9 of the cell free extract showed maximum cobalt uptake for all the investigated cobalt concentrations. In this fraction, glutathione was only induced at 500, 800 and 1000 ppm. Maximum glutathione concentration supported maximum cobalt uptake at 800 ppm. Low molecular weight protein profiles of fraction number 9 revealed that the presence of cobalt induced the appearance of new proteins that were not detected in the same fraction of the control. These low molecular weight peptides (12-5 KDa) suggest the production of Co-metallothioneins. This is the first report of cobalt-induced glutathione by P. brevicompactum and suggests the possible production of phytochelatins.

  14. Cyclopiazonic acid production by Penicillium camemberti Thom and natural occurrence of this mycotoxin in cheese.

    PubMed Central

    Le Bars, J

    1979-01-01

    Every Penicillium camemberti strain freshly isolated from 20 commercial cheese brands produced cyclopiazonic acid in two culture media at 25, 13, and 4 degrees C; the toxin yield was greatly dependent on the strain and environmental parameters (medium, temperature, and incubation time). The toxigenic ability appeared as a log-normal distribution. This mycotoxin was found in the crust (0.05 to 0.1 microgram/g in three samples, 0.1 to 0.2 microgram/g in five samples, and 0.4, 1, and 1.5 microgram/g in three other samples) but not in the inner part. When its acute toxicity is considered, doses eventually ingested by consumers are very low (lower than 4 microgram). Means for prevention are discussed. A highly toxigenic strength and rate appear to be necessary features leading to natural contamination in cheeses. The distribution of toxigenic ability makes possible without delay a choice of weakly toxic strains. PMID:526014

  15. Decolorization of different textile dyes by Penicillium simplicissimum and toxicity evaluation after fungal treatment

    PubMed Central

    Bergsten-Torralba, L.R.; Nishikawa, M.M.; Baptista, D.F.; Magalhães, D.P.; da Silva, M.

    2009-01-01

    The objective of this study was to investigate the capacity of decolorization and detoxification of the textile dyes Reactive Red 198 (RR198), Reactive Blue 214 (RB214), Reactive Blue 21 (RB21) and the mixture of the three dyes (MXD) by Penicillium simplicissimum INCQS 40211. The dye RB21, a phthalocyanine, was totally decolorized in 2 days, and the others, the monoazo RR198, the diazo RB214 and MXD were decolorized after 7 days by P. simplicissimum. Initially the dye decolorization involved dye adsorption by the biomass followed by degradation. The acute toxicity after fungal treatment was monitored with the microcrustacean Daphnia pulex and measured through Effective Concentration 50% (EC50). P. simplicissimum reduced efficiently the toxicity of RB21 from moderately acutely toxic to minor acutely toxic and it also reduced the toxicity of RB214 and MXD, which remained minor acutely toxic. Nevertheless, the fungus increased the toxicity of RR198 despite of the reduction of MXD toxicity, which included this dye. Thus, P. simplicissimum INCQS 40211 was efficient to decolorize different textile dyes and the mixture of them with a significant reduction of their toxicity. In addition this investigation also demonstrated the need of toxicological assays associated to decolorization experiments. PMID:24031428

  16. Influence of intraspecific variability and abiotic factors on mycotoxin production in Penicillium roqueforti.

    PubMed

    Fontaine, Kévin; Hymery, Nolwenn; Lacroix, Marlène Z; Puel, Sylvie; Puel, Olivier; Rigalma, Karim; Gaydou, Vincent; Coton, Emmanuel; Mounier, Jérôme

    2015-12-23

    Penicillium roqueforti has the ability to produce secondary metabolites, including roquefortine C (ROQC) and mycophenolic acid (MPA). In a previous study, the presence of these mycotoxins, alone or in co-occurrence, has been reported in blue-veined cheese. A high variability of mycotoxin content has also been observed, although the majority of samples exhibited relatively low concentrations. The observed variability raises the question of the factors impacting ROQC and MPA production. In this context, the mycotoxigenic potential of 96 P. roqueforti strains (biotic factor) and the effect of some abiotic factors (pH, temperature, NaCl and O2 contents, and C/N ratio) on mycotoxin production were evaluated. A high intraspecific diversity, established via genotypic (RAPD) and phenotypic (FTIR) approaches, was observed. It was associated with mycotoxigenic potential variability and may thus explain part of the observed variability in mycotoxin content of blue-veined cheese. Moreover, a significant decrease of ROQC and MPA production was observed for conditions (temperature, C/N ratio, O2 and NaCl concentrations) encountered during cheese-making compared with optimal growth conditions. The results also highlighted that there was no significant effect of addition of ROQC amino-acid precursor on the production of both mycotoxins whereas a pH increase from 4.5 to 6.5 slightly reduced MPA but not ROQC production.

  17. Statistical optimization of alkaline protease production from Penicillium citrinum YL-1 under solid-state fermentation.

    PubMed

    Xiao, Yun-Zhu; Wu, Duan-Kai; Zhao, Si-Yang; Lin, Wei-Min; Gao, Xiang-Yang

    2015-01-01

    Proteases from halotolerant and halophilic microorganisms were found in traditional Chinese fish sauce. In this study, 30 fungi were isolated from fermented fish sauce in five growth media based on their morphology. However, only one strain, YL-1, which was identified as Penicillium citrinum by internal transcribed spacer (ITS) sequence analysis, can produce alkaline protease. This study is the first to report that a protease-producing fungus strain was isolated and identified in traditional Chinese fish sauce. Furthermore, the culture conditions of alkaline protease production by P. citrinum YL-1 in solid-state fermentation were optimized by response surface methodology. First, three variables including peptone, initial pH, and moisture content were selected by Plackett-Burman design as the significant variables for alkaline protease production. The Box-Behnken design was then adopted to further investigate the interaction effects between the three variables on alkaline protease production and determine the optimal values of the variables. The maximal production (94.30 U/mL) of alkaline protease by P. citrinum YL-1 took place under the optimal conditions of peptone, initial pH, and moisture content (v/w) of 35.5 g/L, 7.73, and 136%, respectively.

  18. [Construction of Producers of Cellulolytic and Pectinolytic Enzymes Based on the Fungus Penicillium verruculosum].

    PubMed

    Bushina, E V; Rubtsova, E A; Rozhkova, A M; Sinitsyna, O A; Koshelev, A V; Matys, V Yu; Nemashkalov, V A; Sinitsyn, A P

    2015-01-01

    Based on the fungus Penicillium verruculosum, we created strains with a complex of extracellular enzymes that contains both cellulolytic enzymes of the fungus and heterologous pectin lyase A from P. canescens and endo- 1,4-α-polygalacturonase from Aspergillus niger. The endopolygalacturonase and pectin lyase activities of enzyme preparations obtained from culture media of the producer strains reached 46-53 U/mg of protein and 1.3-2.3 U/mg of protein, respectively. The optimal temperature and pH values for recombinant pectin lyase and endopolygalacturonase corresponded to those described in the literature for these enzymes. The content of heterologous endopolygalacturonase and pectin lyase in the studied enzyme preparations was 4-5% and 23% of the total protein content, respectively. The yield of reducing sugars upon the hydrolysis of sugar beet and apple processing wastes with the most efficient preparation was 41 and 71 g/L, respectively, which corresponded to a polysaccharide conversion of 49% and 65%. Glucose was the main product of the hydrolysis of sugar beet and apple processing wastes.

  19. Competition during submerged mixed culture of Geotrichum candidum and Penicillium camembertii on glucose and threonine.

    PubMed

    Aziza, Majda; Amrane, Abdeltif

    2008-02-01

    Geotrichum candidum and Penicillium camembertii were cultivated in pure and mixed cultures on glucose and threonine. In pure cultures, G. candidum used glucose as a carbon and an energy source and threonine only as a nitrogen source, even after glucose exhaustion. Contrarily, when growing in isolation, P. camembertii used simultaneously threonine and glucose as carbon sources. A diauxic growth was recorded during the mixed culture of both species, which competed for glucose, the sole carbon source available for G. candidum growth, leading to higher glucose consumption rates than those recorded during pure cultures, while after glucose exhaustion, low growth was recorded in a second step, showing a 'competition' for threonine, the sole remaining carbon and nitrogen sources, confirmed by the increase of 1.0+/-0.1 log of the G. candidum Colony Forming Units. 'Competition' between G. candidum and P. camembertii for the limiting substrate was found to have a positive effect on growth, since it did not lead to the annihilation of one species, as usually observed, but in their coexistence, leading to a rather similar final number of the CFUs for the two populations. 'Competition' resulted in the absence of assimilation of the second available carbon substrate (lactate) as previously observed, or its use only as a nitrogen source, as was the case for threonine in this work.

  20. Three austin family compounds from Penicillium brasilianum exhibit selective blocking action on cockroach nicotinic acetylcholine receptors.

    PubMed

    Kataoka, Saori; Furutani, Shogo; Hirata, Koichi; Hayashi, Hideo; Matsuda, Kazuhiko

    2011-01-01

    Austin (AT) and its derivatives (dehydroaustin (DAT) and acetoxydehydroaustin (ADAT)) produced by Penicillium brasilianum MG-11 exhibit toxicity to insects, yet their targets are unknown. Here, we used whole-cell patch-clamp electrophysiology to investigate the action of AT family compounds on cockroach acetylcholine (ACh), γ-aminobutyric acid (GABA) and l-glutamate receptors expressed in the American cockroach (Periplaneta americana) neuron. U-tube application of AT or its derivatives did not induce any current amplitudes, suggesting that they did not act as agonist of these three receptors. In the second step of experiments, they were bath-applied for 1min before co-application with the corresponding ligand. We found that AT and its derivatives had no effect on GABA and l-glutamate-induced currents, whereas they significantly reduced ACh- and epibatidine-induced currents, showing that these compounds acted as selective antagonists of nicotinic acetylcholine receptors (nAChRs) expressed in the cockroach neuron. Of the compounds, DAT showed the highest blocking potency for nAChRs, differentially attenuating the peak and slowly desensitizing current amplitude of ACh-induced responses with pIC(50) (=-logIC(50) (M)) values of 6.11 and 5.91, respectively. DAT reduced the maximum normalized response to ACh without a significant shift in EC(50), suggesting that the blocking action is not competitive with ACh.

  1. Role of Cannomys badius as a Natural Animal Host of Penicillium marneffei in India

    PubMed Central

    Gugnani, Harish; Fisher, Matthew C.; Paliwal-Johsi, Anubha; Vanittanakom, Nongnuch; Singh, Irabanta; Yadav, Pratap Singh

    2004-01-01

    Infection by Penicillium marneffei in human immunodeficiency virus-positive patients in India has recently been described; the aim of our study was to survey wild rodents and their associated environment in order to identify the natural populations of this fungus. Surveys recovered P. marneffei from the internal organs of 10 (9.1%) of 110 bamboo rats (Cannomys badius) examined from Manipur state, India, an area endemic for penicilliosis marneffei. Identification of the isolates was based on a detailed study of their morphological characteristics, in vitro conversion to fission yeast form, and exoantigen tests. Multilocus microsatellite typing (MLMT) of the isolates revealed five genotypes. No genotypes were shared between sample sites, and all bamboo rats were infected with a single genotype within sample sites, demonstrating spatial genetic heterogeneity. One MLMT genotype was identical to that seen in a human isolate, suggesting that either coinfection from a common source or host-to-host transmission had occurred. This demonstrates the utility of an MLMT-based approach to elucidating the epidemiology of P. marneffei. PMID:15528698

  2. Characterization of sakA gene from pathogenic dimorphic fungus Penicillium marneffei.

    PubMed

    Nimmanee, Panjaphorn; Woo, Patrick C Y; Kummasook, Aksarakorn; Vanittanakom, Nongnuch

    2015-01-01

    Eukaryotes utilize stress activated protein kinase (SAPK) pathways to adapt to environmental stress, including heat, osmotic, oxidative or nutrient stresses. Penicillium marneffei (Talaromyces marneffei), the dimorphic pathogenic fungus that can cause disseminated mycosis in HIV-infected patients, has to encounter various types of stresses both outside and inside host cells. However, the strategies used by this fungus in response to these stresses are still unclear. In this report, the stress-activated kinase (sakA) gene of P. marneffei was characterized and the roles of this gene on various stress conditions were studied. The sakA gene deletion mutant was constructed using the split marker method. The phenotypes and sensitivities to varieties of stresses, including osmotic, oxidative, heat and cell wall stresses of the deletion mutant were compared with the wild type and the sakA complemented strains. Results demonstrated that the P. marneffei sakA gene encoded a putative protein containing TXY phosphorylation lip found in the stress high osmolarity glycerol 1 (Hog1)/Spc1/p38 MAPK family, and that this gene was involved not only in tolerance against oxidative and heat stresses, but also played a role in asexual development, chitin deposition, yeast cell generation in vitro and survival inside mouse and human macrophages.

  3. Penicillium jejuense sp. nov., isolated from the marine environments of Jeju Island, Korea.

    PubMed

    Park, Myung Soo; Fong, Jonathan J; Oh, Seung-Yoon; Houbraken, Jos; Sohn, Jae Hak; Hong, Seung-Beom; Lim, Young Woon

    2015-01-01

    Three strains of an unidentified Penicillium species were isolated during a fungal diversity survey of marine environments in Korea. These strains are described here as a new species following a multigene phylogenetic analyses of nuc rDNA internal transcribed spacer barcodes (ITS1-5.8S-ITS2), genes for β-tubulin, calmodulin and RNA polymerase II second largest subunit, and observation of macro-and micromorphological characters. Phylogenetic analyses revealed that the three strains formed a strongly supported monophyletic group distinct from previously reported species of section Aspergilloides. Morphologically this species can be distinguished from its sister species, P. crocicola, by the reverse color on Czapek yeast autolysate agar, abundant production of sclerotia on malt extract agar and colony characters on yeast extract sucrose agar. We name this new species P. jejuense, after the locality where it was discovered. At 25 C for 7 d, P. jejuense colonies grew to 55-60 mm on CYA, 45-48 mm on MEA, 48-52 mm on YES and 23-26 mm on CREA. Conidia (2.2-3.4 × 2.0-2.6 μm) and sclerotia (160-340 × 125-210 μm) were globose to ellipsoidal.

  4. Inactivation Process of Penicillium digitatum Spores Treated with Non-equilibrium Atmospheric Pressure Plasma

    NASA Astrophysics Data System (ADS)

    Hashizume, Hiroshi; Ohta, Takayuki; Mori, Takumi; Iseki, Sachiko; Hori, Masaru; Ito, Masafumi

    2013-05-01

    To investigate the inactivation process of Penicillium digitatum spores treated with a non-equilibrium atmospheric pressure plasma, the spores were observed using a fluorescent microscope and compared with those treated with ultraviolet (UV) light or moist heat. The treated spores were stained with two fluorescent dyes, 1,1'-dioctadecyl-3,3,Y,3'-tetramethylindocarbocyanine perchlorate (DiI) and diphenyl-1-pyrenylphosphine (DPPP). The intracellular organelles as well as cell membranes in the spores treated with the plasma were stained with DiI without a major morphological change of the membranes, while the organelles were never stained in the spores treated with UV light or moist heat. Moreover, DPPP staining revealed that organelles were oxidized by plasma treatment unlike UV light or moist heat treatments. These results suggest that only plasma treatment induces a minor structural change or functional inhibition of cell membranes, which leads to the oxidation of the intracellular organelles without a major deformation of the membranes through the penetration of reactive oxygen species generated by the plasma into the cell.

  5. 1-Octanol, a self-inhibitor of spore germination in Penicillium camemberti.

    PubMed

    Gillot, Guillaume; Decourcelle, Nicolas; Dauer, Gaëlle; Barbier, Georges; Coton, Emmanuel; Delmail, David; Mounier, Jérôme

    2016-08-01

    Penicillium camemberti is a technologically relevant fungus used to manufacture mold-ripened cheeses. This fungal species produces many volatile organic compounds (VOCs) including ammonia, methyl-ketones, alcohols and esters. Although it is now well known that VOCs can act as signaling molecules, nothing is known about their involvement in P. camemberti lifecycle. In this study, spore germination was shown to be self-regulated by quorum sensing in P. camemberti. This phenomenon, also called "crowding effect", is population-dependent (i.e. observed at high population densities). After determining the volatile nature of the compounds involved in this process, 1-octanol was identified as the main compound produced at high-spore density using GC-MS. Its inhibitory effect was confirmed in vitro and 3 mM 1-octanol totally inhibited spore germination while 100 μM only transiently inhibited spore germination. This is the first time that self-inhibition of spore germination is demonstrated in P. camemberti. The obtained results provide interesting perspectives for better control of mold-ripened cheese processes.

  6. Rare Chromones from a Fungal Mutant of the Marine-Derived Penicillium purpurogenum G59.

    PubMed

    Xia, Ming-Wen; Cui, Cheng-Bin; Li, Chang-Wei; Wu, Chang-Jing; Peng, Ji-Xing; Li, De-Hai

    2015-08-18

    Three new and rare chromones, named epiremisporine B (2), epiremisporine B1 (3) and isoconiochaetone C (4), along with three known remisporine B (1), coniochaetone A (5) and methyl 8-hydroxy-6-methyl-9-oxo-9H-xanthene-1-carboxylate (6) were isolated from a mutant from the diethyl sulfate (DES) mutagenesis of a marine-derived Penicillium purpurogenum G59. The structures of 2-4 including the absolute configurations were determined by spectroscopic methods, especially by NMR analysis and electronic circular dichroism (ECD) experiments in conjunction with calculations. The absolute configuration of the known remisporine B (1) was determined for the first time. Compounds 2 and 3 have a rare feature that has only been reported in one example so far. The compounds 1-6 were evaluated for their cytotoxicity against several human cancer cell lines. The present work explored the great potential of our previous DES mutagenesis strategy for activating silent fungal pathways, which has accelerated the discovery of new bioactive compounds.

  7. The nonylphenol biodegradation study by estuary sediment-derived fungus Penicillium simplicissimum.

    PubMed

    Zhang, Yan; Liu, Ying; Dong, Han; Li, Xianguo; Zhang, Dahai

    2016-08-01

    Nonylphenols (NPs) are persistent organic pollutants (POPs) with estrogenic properties that can perform endocrine-disrupting activities. By using high-concentration NP as environmental selection pressure, one NP biodegradation strain named NPF-4 was isolated and purified from estuary sediment of the Moshui River. It was identified as Penicillium simplicissimum (PS1) by appearance and 18S rDNA analysis. In different culture situations, the strain mass growth and biodegradation ability were evaluated. Within 4-n-nonylphenol (4-n-NP) initial concentration of 20 mg L(-1), it could be degraded 53.76, 90.08, and 100.00 % at 3, 7, and 14 days, respectively. In feeding experiments, it showed that NPF-4 could use 4-n-NP as a sole carbon source. Based on seven products/intermediates detected with GC and LC-MS, a novel biopathway for 4-n-NP biodegradation was proposed, in which sequential hydroxylation, oxidation, and decarboxylation at terminal β-C atom may occur for 4-n-NP detoxification, even complete mineralization in the end.

  8. Biosynthetic gene clusters for relevant secondary metabolites produced by Penicillium roqueforti in blue cheeses.

    PubMed

    García-Estrada, Carlos; Martín, Juan-Francisco

    2016-10-01

    Ripening of blue-veined cheeses, such as the French Bleu and Roquefort, the Italian Gorgonzola, the English Stilton, the Danish Danablu or the Spanish Cabrales, Picón Bejes-Tresviso, and Valdeón, requires the growth and enzymatic activity of the mold Penicillium roqueforti, which is responsible for the characteristic texture, blue-green spots, and aroma of these types of cheeses. This filamentous fungus is able to synthesize different secondary metabolites, including andrastins, mycophenolic acid, and several mycotoxins, such as roquefortines C and D, PR-toxin and eremofortins, isofumigaclavines A and B, and festuclavine. This review provides a detailed description of the main secondary metabolites produced by P. roqueforti in blue cheese, giving a special emphasis to roquefortine, PR-toxin and mycophenolic acid, and their biosynthetic gene clusters and pathways. The knowledge of these clusters and secondary metabolism pathways, together with the ability of P. roqueforti to produce beneficial secondary metabolites, is of interest for commercial purposes.

  9. Allosteric Inhibition via R-state Destabilization in ATP Sulfurylase from Penicillium chrysogenum

    SciTech Connect

    MacRae, I. J.

    2002-01-01

    The structure of the cooperative hexameric enzyme ATP sulfurylase from Penicillium chrysogenum bound to its allosteric inhibitor, 3'-phosphoadenosine-5'-phosphosulfate (PAPS), was determined to 2.6 {angstrom} resolution. This structure represents the low substrate-affinity T-state conformation of the enzyme. Comparison with the high substrate-affinity R-state structure reveals that a large rotational rearrangement of domains occurs as a result of the R-to-T transition. The rearrangement is accompanied by the 17 {angstrom} movement of a 10-residue loop out of the active site region, resulting in an open, product release-like structure of the catalytic domain. Binding of PAPS is proposed to induce the allosteric transition by destabilizing an R-state-specific salt linkage between Asp 111 in an N-terminal domain of one subunit and Arg 515 in the allosteric domain of a trans-triad subunit. Disrupting this salt linkage by site-directed mutagenesis induces cooperative inhibition behavior in the absence of an allosteric effector, confirming the role of these two residues.

  10. Toxicity of penicillic acid for rat alveolar macrophages in vitro. [Aspergillus; Penicillium

    SciTech Connect

    Sorenson, W.G.; Simpson, J.

    1985-12-01

    Penicillic acid (PA) is a polyketide mycotoxin produced by several species of Aspergillus and Penicillium. This mycotoxin is toxic in experimental animals and has also been reported to be carcinogenic. The cytotoxicity of penicillic acid was studied in rat albeolar macrophages (AM) in vitro. The effects of penicillic acid on membrane integrity were studied by measuring cell volume changes and /sup 51/Cr release. There was a significant decrease in adenosine triphosphate (ATP) in cell cultures exposed to 1.0 mM penicillic acid for 4 hr. Inhibition of the incorporation of (/sup 3/H)leucine into protein was both dose- and time-dependent and protein synthesis was inhibited significantly after 2 hr exposure to greater than or equal to0.1 mM penicillic acid. RNA synthesis was inhibited to a lesser extent than protein synthesis. There was significant inhibition of phagocytosis after 2 hr exposure at greater than or equal to0.3 mM penicillic acid and the ED/sub 50/ for phagocytosis was 0.09 mM. Thus phagocytosis was more sensitive to the toxic effects of penicillic acid than any other cellular process studied. The data suggest the possibility of a respiratory hazard to agricultural workers exposed to contaminated grain.

  11. Green synthesis of silver nanoparticle by Penicillium purpurogenum NPMF: the process and optimization

    NASA Astrophysics Data System (ADS)

    Nayak, Rati Ranjan; Pradhan, Nilotpala; Behera, Debadhyan; Pradhan, Kshyama Madhusikta; Mishra, Srabani; Sukla, Lala Behari; Mishra, Barada Kanta

    2011-08-01

    An eco-friendly microbial method for synthesis of silver colloid solution with antimicrobial activity is developed using a fungal strain of Penicillium purpurogenum NPMF. It is observed that increase in concentration of AgNO3 increases the formation of silver nanoparticle. At 5 mM concentration highly populated polydispersed nanoparticles form. Furthermore, change in pH of the reaction mixture leads to change in shape and size of silver nanoparticles. At lower pH two peaks are observed in the absorption spectra showing polydispersity of nanoparticles. However, highly monodispersed spherical nanoparticles of 8-10 nm size form with 1 mM AgNO3 concentration at pH 8. Antimicrobial activity of nanoparticles is demonstrated against pathogenic gram negative bacteria like Escherichia coli and Pseudomonas aeruginosa, and gram positive bacteria like Staphylococcus aureus. The antimicrobial activity of silver nanoparticles obtained at different initial pH show strong dependence on the surface area and shape of the nanoparticles.

  12. Identification and Functional Analysis of the Mycophenolic Acid Gene Cluster of Penicillium roqueforti.

    PubMed

    Del-Cid, Abdiel; Gil-Durán, Carlos; Vaca, Inmaculada; Rojas-Aedo, Juan F; García-Rico, Ramón O; Levicán, Gloria; Chávez, Renato

    2016-01-01

    The filamentous fungus Penicillium roqueforti is widely known as the ripening agent of blue-veined cheeses. Additionally, this fungus is able to produce several secondary metabolites, including the meroterpenoid compound mycophenolic acid (MPA). Cheeses ripened with P. roqueforti are usually contaminated with MPA. On the other hand, MPA is a commercially valuable immunosuppressant. However, to date the molecular basis of the production of MPA by P. roqueforti is still unknown. Using a bioinformatic approach, we have identified a genomic region of approximately 24.4 kbp containing a seven-gene cluster that may be involved in the MPA biosynthesis in P. roqueforti. Gene silencing of each of these seven genes (named mpaA, mpaB, mpaC, mpaDE, mpaF, mpaG and mpaH) resulted in dramatic reductions in MPA production, confirming that all of these genes are involved in the biosynthesis of the compound. Interestingly, the mpaF gene, originally described in P. brevicompactum as a MPA self-resistance gene, also exerts the same function in P. roqueforti, suggesting that this gene has a dual function in MPA metabolism. The knowledge of the biosynthetic pathway of MPA in P. roqueforti will be important for the future control of MPA contamination in cheeses and the improvement of MPA production for commercial purposes.

  13. Identification and functional analysis of Penicillium digitatum genes putatively involved in virulence towards citrus fruit.

    PubMed

    López-Pérez, Mario; Ballester, Ana-Rosa; González-Candelas, Luis

    2015-04-01

    The fungus Penicillium digitatum, the causal agent of green mould rot, is the most destructive post-harvest pathogen of citrus fruit in Mediterranean regions. In order to identify P. digitatum genes up-regulated during the infection of oranges that may constitute putative virulence factors, we followed a polymerase chain reaction (PCR)-based suppression subtractive hybridization and cDNA macroarray hybridization approach. The origin of expressed sequence tags (ESTs) was determined by comparison against the available genome sequences of both organisms. Genes coding for fungal proteases and plant cell wall-degrading enzymes represent the largest categories in the subtracted cDNA library. Northern blot analysis of a selection of P. digitatum genes, including those coding for proteases, cell wall-related enzymes, redox homoeostasis and detoxification processes, confirmed their up-regulation at varying time points during the infection process. Agrobacterium tumefaciens-mediated transformation was used to generate knockout mutants for two genes encoding a pectin lyase (Pnl1) and a naphthalene dioxygenase (Ndo1). Two independent P. digitatum Δndo1 mutants were as virulent as the wild-type. However, the two Δpnl1 mutants analysed were less virulent than the parental strain or an ectopic transformant. Together, these results provide a significant advance in our understanding of the putative determinants of the virulence mechanisms of P. digitatum.

  14. UVA Photoactivation of Harmol Enhances Its Antifungal Activity against the Phytopathogens Penicillium digitatum and Botrytis cinerea.

    PubMed

    Olmedo, Gabriela M; Cerioni, Luciana; González, María M; Cabrerizo, Franco M; Volentini, Sabrina I; Rapisarda, Viviana A

    2017-01-01

    Phytopathogenic fungi responsible for post-harvest diseases on fruit and vegetables cause important economic losses. We have previously reported that harmol (1-methyl-9H-pyrido[3,4-b]indol-7-ol) is active against the causal agents of green and gray molds Penicillium digitatum and Botrytis cinerea, respectively. Here, antifungal activity of harmol was characterized in terms of pH dependency and conidial targets; also photodynamic effects of UVA irradiation on the antimicrobial action were evaluated. Harmol was able to inhibit the growth of both post-harvest fungal disease agents only in acidic conditions (pH 5), when it was found in its protonated form. Conidia treated with harmol exhibited membrane integrity loss, cell wall disruption, and cytoplasm disorganization. All these deleterious effects were more evident for B. cinerea in comparison to P. digitatum. When conidial suspensions were irradiated with UVA in the presence of harmol, antimicrobial activity against both pathogens was enhanced, compared to non-irradiated conditions. B. cinerea exhibited a high intracellular production of reactive oxygen species (ROS) when was incubated with harmol in irradiated and non-irradiated treatments. P. digitatum showed a significant increase in ROS accumulation only when treated with photoexcited harmol. The present work contributes to unravel the antifungal activity of harmol and its photoexcited counterpart against phytopathogenic conidia, focusing on ROS accumulation which could account for damage on different cellular targets.

  15. Antifungal activity of β-carbolines on Penicillium digitatum and Botrytis cinerea.

    PubMed

    Olmedo, Gabriela M; Cerioni, Luciana; González, M Micaela; Cabrerizo, Franco M; Rapisarda, Viviana A; Volentini, Sabrina I

    2017-04-01

    β-carbolines (βCs) are alkaloids widely distributed in nature that have demonstrated antimicrobial properties. Here, we tested in vitro six βCs against Penicillium digitatum and Botrytis cinerea, causal agents of postharvest diseases on fruit and vegetables. Full aromatic βCs (harmine, harmol, norharmane and harmane) exhibited a marked inhibitory effect on conidia germination at concentrations between 0.5 and 1 mM, while dihydro-βCs (harmalina and harmalol) only caused germination delay. Harmol showed the highest inhibitory effect on both fungal pathogens. After 24 h of exposure to 1 mM harmol, conidia revealed a severe cellular damage, exhibiting disorganized cytoplasm and thickened cell wall. Harmol antimicrobial effect was fungicidal on B. cinerea, while it was fungistatic on P. digitatum. Conidia membrane permeabilization was detected in treatments with harmol at sub-inhibitory and inhibitory concentrations, for both pathogens. In addition, residual infectivity of P. digitatum on lemons and B. cinerea on blueberries was significantly reduced after exposure to this alkaloid. It also inhibited mycelial growth, preventing sporulation at the highest concentration tested. These results indicate that harmol might be a promising candidate as a new antifungal molecule to control causal agents of fruit diseases.

  16. Integrated control of Penicillium digitatum by the predacious yeast Saccharomycopsis crataegensis and sodium bicarbonate on oranges

    PubMed Central

    Pimenta, R. S.; Silva, J. F. M.; Coelho, C. M.; Morais, P. B.; Rosa, C. A.; Corrêa Jr, A.

    2010-01-01

    Our investigation of integrated biological control (IBC) started with an assay testing activity of the predacious yeast Saccharomycopsis crataegensis UFMG-DC19.2 against Penicillium digitatum LCP 4354, a very aggressive fungus that causes postharvest decay in oranges. Under unfavourable environmental conditions, the yeast showed a high potential for control (39.9% disease severity reduction) of this fungus. This result was decisive for the next step, in which S. crataegensis was tested in association with sodium bicarbonate salt, a generally regarded as safe (GRAS) substance. The yeast was able to survive at different concentrations of the salt (1%, 2% and 5%), and continued to grow for a week at the wound site, remaining viable at high population for 14 days on the fruit surface. The yeast alone reduced the severity of decay by 41.7% and sodium bicarbonate alone reduced severity of decay by 19.8%, whereas the application of both led to a delay in the development of symptoms from 2 to 10 days. Ingredients of the formulations were not aggressive to fruits since no lesions were produced in control experiments. PMID:24031511

  17. Bioethanol potentials of corn cob hydrolysed using cellulases of Aspergillus niger and Penicillium decumbens

    PubMed Central

    Saliu, Bolanle Kudirat; Sani, Alhassan

    2012-01-01

    Corn cob is a major component of agricultural and domestic waste in many parts of the world. It is composed mainly of cellulose which can be converted to energy in form of bioethanol as an efficient and effective means of waste management. Production of cellulolytic enzymes were induced in the fungi Aspergillus niger and Penicillium decumbens by growing them in mineral salt medium containing alkali pre-treated and untreated corn cobs. The cellulases were characterized and partially purified. Alkali pre-treated corn cobs were hydrolysed with the partially purified cellulases and the product of hydrolysis was fermented using the yeast saccharomyces cerevisae to ethanol. Cellulases of A. niger produced higher endoglucanase and exoglucanase activity (0.1698 IU ml-1 and 0.0461 FPU ml-1) compared to that produced by P. decumbens (0.1111 IU ml-1 and 0.153 FPU ml-1). Alkali pre-treated corn cob hydrolysed by cellulases of A. niger yielded 7.63 mg ml-1 sugar which produced 2.67 % (v/v) ethanol on fermentation. Ethanol yield of the hydrolysates of corn cob by cellulases of P. decumbens was much lower at 0.56 % (v/v). Alkali pre-treated corn cob, hydrolysed with cellulases of A. niger is established as suitable feedstock for bioethanol production. PMID:27418920

  18. UP-HILIC-MS/MS to Determine the Action Pattern of Penicillium sp. Dextranase

    NASA Astrophysics Data System (ADS)

    Yi, Lin; Sun, Xue; Du, Kenze; Ouyang, Yilan; Wu, Chengling; Xu, Naiyu; Linhardt, Robert J.; Zhang, Zhenqing

    2015-07-01

    Investigation of the action pattern of enzymes acting on carbohydrates is challenging, as both the substrate and the digestion products are complex mixtures. Dextran and its enzyme-derived oligosaccharides are widely used for many industrial applications. In this work, a new method relying on ultra-performance hydrophilic interaction liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UP-HILIC- Q/TOF-MS/MS) was developed to analyze a complex mixture of dextran oligosaccharide products to determine the action pattern of dextranase. No derivatization of oligosaccharides was required and the impact of the α- and β-configurations of the native oligosaccharides on the chromatographic separation was eliminated. The 1→6, 1→3, 1→4 backbone linkages and the branch linkages of these oligosaccharides were all distinguished from diagnostic ions in their MS/MS spectra, including fragments corresponding to 0,2A, 0,3A, 0,4A, B-H2O, 2,5A, and 3,5A. The sequences of the oligosaccharide products were similarly established. Thus, the complex oligosaccharide mixtures in dextran digestion products were profiled and identified using this method. The more enzyme-resistant structures in dextran were established using much less sample, labor, time, and uncertainty than in previous studies. This method provides an efficient, sensitive, and straightforward way to monitor the entire process of digestion, establish the action pattern of the dextranase from Penicillium sp., and to support the proper industrial application of dextranase.

  19. Neurotoxicity of mycotoxins produced in vitro by Penicillium roqueforti isolated from maize and grass silage.

    PubMed

    Malekinejad, H; Aghazadeh-Attari, J; Rezabakhsh, A; Sattari, M; Ghasemsoltani-Momtaz, B

    2015-10-01

    Fungal growth in human foods and animal feeds causes profound damage indicating a general spoilage, nutritional losses, and the formation of mycotoxins. Thirty apparently contaminated maize and grass silage samples were analyzed for the presence of total fungi. Penicillium roqueforti were isolated from all (100%) moldy silage samples on general and selective culture media. Furthermore, P. roqueforti-positive samples culture media subjected to the toxin extraction and toxins of patulin, penicillic acid, mycophenolic acid, and roquefortin-C (ROQ-C) were identified by means of high-performance liquid chromatography method. Cytotoxicity of identified toxins was investigated on neuro-2a cells. Alamar blue reduction, neutral red uptake, and intracellular adenosine triphosphate (ATP) content assays indicated that patulin and ROQ-C exert the strongest and weakest toxicity, respectively. Reactive oxygen species (ROS) generation by the toxins-exposed cells was measured, and the results supported the mitochondrial and lysosomal dysfunction and ATP depletion in exposed cells. Our data suggest that P. roqueforti is the widely present mold in analyzed maize and grass silage samples, which is able to produce toxins that cause neurotoxicity. This finding may explain in part some neuronal disorders in animals, which are fed contaminated feedstuffs with mentioned fungus. Moreover, mitochondrial and lysosomal dysfunction, intracellular ATP depletion, and the excessive ROS generation were found as the mechanisms of cytotoxicity for P. roqueforti-produced toxins.

  20. Integrated control of Penicillium digitatum by the predacious yeast Saccharomycopsis crataegensis and sodium bicarbonate on oranges.

    PubMed

    Pimenta, R S; Silva, J F M; Coelho, C M; Morais, P B; Rosa, C A; Corrêa, A

    2010-04-01

    Our investigation of integrated biological control (IBC) started with an assay testing activity of the predacious yeast Saccharomycopsis crataegensis UFMG-DC19.2 against Penicillium digitatum LCP 4354, a very aggressive fungus that causes postharvest decay in oranges. Under unfavourable environmental conditions, the yeast showed a high potential for control (39.9% disease severity reduction) of this fungus. This result was decisive for the next step, in which S. crataegensis was tested in association with sodium bicarbonate salt, a generally regarded as safe (GRAS) substance. The yeast was able to survive at different concentrations of the salt (1%, 2% and 5%), and continued to grow for a week at the wound site, remaining viable at high population for 14 days on the fruit surface. The yeast alone reduced the severity of decay by 41.7% and sodium bicarbonate alone reduced severity of decay by 19.8%, whereas the application of both led to a delay in the development of symptoms from 2 to 10 days. Ingredients of the formulations were not aggressive to fruits since no lesions were produced in control experiments.

  1. Comparative study of antimicrobial peptides to control citrus postharvest decay caused by Penicillium digitatum.

    PubMed

    Muñoz, Alberto; López-García, Belén; Marcos, Jose F

    2007-10-03

    The objective of this study was to investigate and compare the in vitro efficacy and in vivo potential of eight distinct short antimicrobial peptides to control the postharvest green mold disease of oranges caused by the fungus Penicillium digitatum. The L-amino acid versions of the four peptides PAF26, PAF38, PAF40, and BM0, previously obtained by combinatorial approaches, were examined. The study included two antibacterial peptides formerly identified by rational design, BP15 and BP76, and it is demonstrated that they also have in vitro antifungal properties. The natural antimicrobial peptides melittin and indolicidin were also selected for comparison, due to their well-known properties and modes of action. In vitro and in vivo results indicated differential behaviors among peptides, regarding the inhibitory potency in growth media, selectivity against distinct microorganisms, fungicidal activity towards nongerminated conidia of P. digitatum, and efficacy in fruit inoculation assays. Interestingly, a high in vitro inhibitory activity did not necessarily associate with an effective control of fruit infection by P. digitatum. The short tryptophan-rich cationic peptides PAF26, PAF38, PAF40, and BM0 were lethal to conidia of P. digitatum, and this property is correlated with better protection in the decay control test.

  2. Penicillium menonorum: A Novel Fungus to Promote Growth and Nutrient Management in Cucumber Plants

    PubMed Central

    Babu, Anam Giridhar; Kim, Sang Woo; Yadav, Dil Raj; Hyum, Umyong; Adhikari, Mahesh

    2015-01-01

    The present study is the first report on the isolation of Penicillium menonorum from rhizosphere soil in Korea and its identification based on morphological characteristics and internal transcribed spacer gene sequence. The fungal isolate was named KNU-3 and was found to exhibit plant growth-promoting (PGP) activity through indole acetic acid (IAA) and siderophore production, as well as P solubilization. KNU-3 produced 9.7 mg/L IAA and solubilized 408 mg of Ca3PO4/L, and inoculation with the isolate significantly (p < 0.05) increased the dry biomass of cucumber roots (57%) and shoots (52%). Chlorophyll, starch, protein, and P contents were increased by 16%, 45%, 22%, and 14%, respectively, compared to plants grown in uninoculated soil. The fungus also increased soil dehydrogenase (30%) and acid phosphatase (19%) activities. These results demonstrate that the isolate KNU-3 has potential PGP attributes, and therefore it can be considered as a new fungus to enhance soil fertility and promote plant growth. Moreover, the discovery of PGP ability and traits of this fungus will open new aspects of research and investigations. In this study, plant growth promotion by P. menonorum KNU-3 is reported for the first time in Korea after its original description. PMID:25892915

  3. Assessment of exposure to the Penicillium glabrum complex in cork industry using complementing methods.

    PubMed

    Viegas, Carla; Sabino, Raquel; Botelho, Daniel; dos Santos, Mateus; Gomes, Anita Quintal

    2015-09-01

    Cork oak is the second most dominant forest species in Portugal and makes this country the world leader in cork export. Occupational exposure to Chrysonilia sitophila and the Penicillium glabrum complex in cork industry is common, and the latter fungus is associated with suberosis. However, as conventional methods seem to underestimate its presence in occupational environments, the aim of our study was to see whether information obtained by polymerase chain reaction (PCR), a molecular-based method, can complement conventional findings and give a better insight into occupational exposure of cork industry workers. We assessed fungal contamination with the P. glabrum complex in three cork manufacturing plants in the outskirts of Lisbon using both conventional and molecular methods. Conventional culturing failed to detect the fungus at six sampling sites in which PCR did detect it. This confirms our assumption that the use of complementing methods can provide information for a more accurate assessment of occupational exposure to the P. glabrum complex in cork industry.

  4. Extracellular inulinases from Penicillium janczewskii, a fungus isolated from the rhizosphere of Vernonia herbacea (Asteraceae).

    PubMed

    Pessoni, R A; Figueiredo-Ribeiro, R C; Braga, M R

    1999-07-01

    Extracellular inulinases from Penicillium janczewskii were obtained from the filtrate of 12 day-old cultures supplemented with inulin from Vernonia herbacea. Crude filtrates and partially-purified enzyme preparations (peaks I and II) were active on inulin, sucrose and raffinose. The apparent M(r) of the enzymes from peaks I and II were 48 and 66 kDa, respectively. The apparent K(m) (mmol l-1) values of peak I were 0.43 for inulin and 18.7 for sucrose; for peak II they were 0.87 and 18.5 for inulin and sucrose, respectively. Their temperature and pH optima were 55 degrees C and 5.0, respectively. Both peaks catalysed the hydrolysis of beta-(2,1) fructans more rapidly than beta-(2,6) fructans. Free fructose was the predominant product released from inulin, indicating that these enzymes display exo-inulinase activity. In view of these characteristics, the yield and the high specific activity towards beta-(2,1) fructans, inulinases from P. janczewskii can be utilized for the preparation of fructose syrup from inulin.

  5. Fertility depression among cheese‐making Penicillium roqueforti strains suggests degeneration during domestication

    PubMed Central

    Ropars, Jeanne; Lo, Ying‐Chu; Dumas, Emilie; Snirc, Alodie; Begerow, Dominik; Rollnik, Tanja; Lacoste, Sandrine; Dupont, Joëlle; Giraud, Tatiana; López‐Villavicencio, Manuela

    2016-01-01

    Genetic differentiation occurs when gene flow is prevented, due to reproductive barriers or asexuality. Investigating the early barriers to gene flow is important for understanding the process of speciation. Here, we therefore investigated reproductive isolation between different genetic clusters of the fungus Penicillium roqueforti, used for maturing blue cheeses, and also occurring as food spoiler or in silage. We investigated premating and postmating fertility between and within three genetic clusters (two from cheese and one from other substrates), and we observed sexual structures under scanning electron microscopy. All intercluster types of crosses showed some fertility, suggesting that no intersterility has evolved between domesticated and wild populations despite adaptation to different environments and lack of gene flow. However, much lower fertility was found in crosses within the cheese clusters than within the noncheese cluster, suggesting reduced fertility of cheese strains, which may constitute a barrier to gene flow. Such degeneration may be due to bottlenecks during domestication and/or to the exclusive clonal replication of the strains in industry. This study shows that degeneration has occurred rapidly and independently in two lineages of a domesticated species. Altogether, these results inform on the processes and tempo of degeneration and speciation. PMID:27470007

  6. Overexpression of the plg1 gene encoding pectin lyase in Penicillium griseoroseum.

    PubMed

    Cardoso, Patrícia Gomes; Ribeiro, João Batista; Teixeira, Janaina Aparecida; de Queiroz, Marisa Vieira; de Araújo, Elza Fernandes

    2008-03-01

    The pectin lyase (PL) is an industrially important enzyme since it is used for maceration and clarification in the process of fruit juice production in food industries. In order to increase the yields of pectin lyase we cloned the plg1 (pectin lyase 1) from Penicillium griseoroseum gene under the control of the strong constitutive promoter of the glyceraldehyde-3-phosphate dehydrogenase gene (gpdA) and the terminator region of the tryptophan synthetase (trpC) gene from Aspergillus nidulans (plasmid pAN52-Plg1) and transformed this construct into the P. griseoroseum strain PG63. One of the pAN52-Plg1 multi-copy transformants (strain 105) grown in culture medium containing glucose or sugar cane juice showed PL activities of 4,804 or 5,202 U ml(-1) respectively, which represented 57- and 132-fold increases. In addition, the apparent specific activity of PL produced by this strain was much higher than the one observed for a commercial pectinase preparation. Evaluation of the extracellular proteins in the culture supernatant of strain 105 by SDS-PAGE showed the presence of a clear and strong band of approximately 40 kDa that probably corresponds to PL. The enzyme yields reported here demonstrate that the system we developed is able to express pectin lyase at levels comparable to, or exceeding, previously reported data.

  7. Role of Cannomys badius as a natural animal host of Penicillium marneffei in India.

    PubMed

    Gugnani, Harish; Fisher, Matthew C; Paliwal-Johsi, Anubha; Vanittanakom, Nongnuch; Singh, Irabanta; Yadav, Pratap Singh

    2004-11-01

    Infection by Penicillium marneffei in human immunodeficiency virus-positive patients in India has recently been described; the aim of our study was to survey wild rodents and their associated environment in order to identify the natural populations of this fungus. Surveys recovered P. marneffei from the internal organs of 10 (9.1%) of 110 bamboo rats (Cannomys badius) examined from Manipur state, India, an area endemic for penicilliosis marneffei. Identification of the isolates was based on a detailed study of their morphological characteristics, in vitro conversion to fission yeast form, and exoantigen tests. Multilocus microsatellite typing (MLMT) of the isolates revealed five genotypes. No genotypes were shared between sample sites, and all bamboo rats were infected with a single genotype within sample sites, demonstrating spatial genetic heterogeneity. One MLMT genotype was identical to that seen in a human isolate, suggesting that either coinfection from a common source or host-to-host transmission had occurred. This demonstrates the utility of an MLMT-based approach to elucidating the epidemiology of P. marneffei.

  8. Isolation and analysis of differentially expressed genes in Penicillium glabrum subjected to thermal stress.

    PubMed

    Nevarez, L; Vasseur, V; Le Dréan, G; Tanguy, A; Guisle-Marsollier, I; Houlgatte, R; Barbier, G

    2008-12-01

    Penicillium glabrum is a filamentous fungus frequently involved in food contamination. Numerous environmental factors (temperature, humidity, atmospheric composition, etc.) or food characteristics (water activity, pH, preservatives, etc.) could represent potential sources of stress for micro-organisms. These factors can directly affect the physiology of these spoilage micro-organisms: growth, conidiation, synthesis of secondary metabolites, etc. This study investigated the transcriptional response to temperature in P. glabrum, since this factor is one of the most important for fungal growth. Gene expression was first analysed by using suppression subtractive hybridization to generate two libraries containing 445 different up- and downregulated expressed sequence tags (ESTs). Expression of these ESTs was then assessed for different thermal stress conditions, with cDNA microarrays, resulting in the identification of 35 and 49 significantly up- and downregulated ESTs, respectively. These ESTs encode heat-shock proteins, ribosomal proteins, superoxide dismutase, trehalose-6-phosphate synthase and a large variety of identified or unknown proteins. Some of these may be molecular markers for thermal stress response in P. glabrum. To our knowledge, this work represents the first study of the transcriptional response of a food spoilage filamentous fungus under thermal stress conditions.

  9. Identification of a GH62 α-L-arabinofuranosidase specific for arabinoxylan produced by Penicillium chrysogenum.

    PubMed

    Sakamoto, Tatsuji; Ogura, Atsuhiro; Inui, Misako; Tokuda, Sayaka; Hosokawa, Sachiko; Ihara, Hideshi; Kasai, Naoya

    2011-04-01

    An arabinoxylan arabinofuranohydrolase (AXS5) was purified from the culture filtrate of Penicillium chrysogenum 31B. A cDNA encoding AXS5 (axs5) was isolated by in vitro cloning using the N-terminal amino acid sequence of the native enzyme as a starting point. The deduced amino acid sequence of the axs5 gene has high similarities with those of arabinoxylan arabinofuranohydrolases of Aspergillus niger, Aspergillus tubingensis, and Aspergillus sojae. Module sequence analysis revealed that a "Glyco_hydro_62" was present at position 28-299 of AXS5. This is a family of α-L-arabinofuranosidases which are all members of glycoside hydrolase family 62. Recombinant AXS5 (rAXS5) expressed in Escherichia coli was highly active on arabinoxylan but not on branched sugar beet arabinan. (1)H-NMR analysis revealed that the rAXS5 cleaved arabinosyl side-chains linked to C-2 and C-3 of single-substituted xylose residues in arabinoxylan. Semi-quantitative RT-PCR analysis indicated that expression of the axs5 gene in P. chrysogenum 31B was strongly induced by adding D-xylose and arabinoxylan to the culture medium. Moreover, two binding sites of XlnR, a transcriptional activator that regulates the expression of the genes encoding xylanolytic enzymes, are present in the upstream region of the axs5 gene. These results suggest that AXS5 is involved in xylan degradation.

  10. Identification and characterization of an acidic and acid-stable endoxyloglucanase from Penicillium oxalicum.

    PubMed

    Xian, Liang; Wang, Fei; Yin, Xin; Feng, Jia-Xun

    2016-05-01

    Xyloglucan is a major structural macromolecule of the primary cell wall of spermatophytes. The hydrolysis of xyloglucan by xyloglucanases may facilitate the hydrolysis of cellulose by cellulases, which is beneficial for bioethanol production. Penicillium oxalicum has been employed for commercial cellulase production. In P. oxalicum, many genes and proteins related to the degradation of structural macromolecules of the plant cell wall have been found, but no gene encoding a xyloglucanase has been identified. In this study, a gene, PoxXEG12A, was cloned from P. oxalicum and expressed in Pichia pastoris, and the gene product was enzymatically characterized. PoxXEG12A shared 63% sequence identity with endoxyloglucanases from Aspergillus niger and Aspergillus aculeatus. PoxXEG12A specifically hydrolyzed tamarind xyloglucan in endo-acting mode and, thus, it is an endoxyloglucanase. PoxXEG12A was most active at pH 4.5-5.5 and at 55-60 °C, with a specific activity of 172 U/mg protein toward tamarind xyloglucan. The enzyme was stable at pH 3.5-7.0 and below 40 °C. The properties of the endoxyloglucanase PoxXEG12A suggest that the enzyme might have potential in industrial applications.

  11. Detection of griseofulvin in a marine strain of Penicillium waksmanii by ion trap mass spectrometry.

    PubMed

    Petit, K E; Mondeguer, F; Roquebert, M F; Biard, J F; Pouchus, Y F

    2004-07-01

    A marine strain of Penicillium waksmanii Zaleski was isolated from a sample of seawater from shellfish-farming area in the Loire estuary (France). The in vitro marine culture showed an important antifungal activity. Bioassay-guided fractionation was used to purify the crude extract. Dereplication by electrospray-ion trap/mass spectrometry (ESI-IT/MS) afforded the identification of the antifungal compound, after a semi-purification consisting of two stages. A comparison of the ionic composition between the active and the non-active fractions allowed the detection of a monocharged ion at m/z 353 containing a chlorine atom, which could be attributed to the antifungal griseofulvin [C17H17ClO6+H]+. Multi-stage fragmentation (MSn) confirmed the identity of the m/z 353 ion of the antifungal fraction as griseofulvin. It is the first description of griseofulvin production by a strain of P. waksmanii and the first chemical study of a strain of this species isolated from marine temperate cold water.

  12. Identification and functional characterization of Penicillium marneffei pleiotropic drug resistance transporters ABC1 and ABC2.

    PubMed

    Panapruksachat, Siribun; Iwatani, Shun; Oura, Takahiro; Vanittanakom, Nongnuch; Chindamporn, Ariya; Niimi, Kyoko; Niimi, Masakazu; Lamping, Erwin; Cannon, Richard D; Kajiwara, Susumu

    2016-07-01

    Penicilliosis caused by the dimorphic fungus Penicillium marneffei is an endemic, AIDS-defining illness and, after tuberculosis and cryptococcosis, the third most common opportunistic infection of AIDS patients in tropical Southeast Asia. Untreated, patients have poor prognosis; however, primary amphotericin B treatment followed by prolonged itraconazole prophylaxis is effective. To identify ATP-binding cassette (ABC) transporters that may play a role in potential multidrug resistance of P. marneffei, we identified and classified all 46 P. marneffei ABC transporters from the genome sequence. PmABC1 and PmABC2 were most similar to the archetype Candida albicans multidrug efflux pump gene CDR1. P. marneffei Abc1p (PmAbc1p) was functionally expressed in Saccharomyces cerevisiae, although at rather low levels, and correctly localized to the plasma membrane, causing cells to be fourfold to eightfold more resistant to azoles and many other xenobiotics than untransformed cells. P. marneffei Abc2p (PmAbc2p) was expressed at similarly low levels, but it had no efflux activity and did not properly localize to the plasma membrane. Interestingly, PmAbc1p mislocalized and lost its transport activity when cells were shifted to 37 °C. We conclude that expression of PmAbc1p in S. cerevisiae confers resistance to several xenobiotics indicating that PmAbc1p may be a multidrug efflux pump.

  13. Comparison of Free and Immobilized L-asparaginase Synthesized by Gamma-Irradiated Penicillium cyclopium.

    PubMed

    El-Refai, Heba A; Shafei, Mona S; Mostafa, Hanan; El-Refai, Abdel-Monem H; Araby, Eman M; El-Beih, Fawkia M; Easa, Saadia M; Gomaa, Sanaa K

    2016-01-01

    Gamma irradiation is used on Penicillium cyclopium in order to obtain mutant cells of high L-asparaginase productivity. Using gamma irradiation dose of 4 KGy, P. cyclopium cells yielded L-asparaginase with extracellular enzyme activity of 210.8 ± 3 U/ml, and specific activity of 752.5 ± 1.5 U/mg protein, which are 1.75 and 1.53 times, respectively, the activity of the wild strain. The enzyme was partially purified by 40-60% acetone precipitation. L-asparaginase was immobilized onto Amberlite IR-120 by ionic binding. Both free and immobilized enzymes exhibited maximum activity at pH 8 and 40 degrees C. The immobilization process improved the enzyme thermal stability significantly. The immobilized enzyme remained 100% active at temperatures up to 60 degrees C, while the free asparaginase was less tolerant to high temperatures. The immobilized enzyme was more stable at pH 9.0 for 50 min, retaining 70% of its relative activity. The maximum reaction rate (V(max)) and Michaelis-Menten constant (K(m)) of the free form were significantly changed after immobilization. The K(m) value for immobilized L-asparaginase was about 1.3 times higher than that of free enzyme. The ions K+, Ba2+ and Na+ showed stimulatory effect on enzyme activity with percentages of 110%, 109% and 106% respectively.

  14. Influence of rhamnolipids and Triton X-100 on adsorption of phenol by Penicillium simplicissimum.

    PubMed

    Liu, Zhifeng; Zeng, Zhuotong; Zeng, Guangming; Li, Jianbing; Zhong, Hua; Yuan, Xingzhong; Liu, Yang; Zhang, Jiachao; Chen, Ming; Liu, Yuanyuan; Xie, Gengxin

    2012-04-01

    The effects of rhamnolipids and Triton X-100 on phenol adsorption by Penicillium simplicissimum were studied. The optimum pH was 7 for phenol adsorption by all the test biomasses. The adsorption of phenol at pH 7 by biomass pre-treated with 0.05% Triton X-100, 0.2% Triton X-100, 0.05% rhamnolipids and 0.005% rhamnolipids was 3.4, 2.7, 2.4, and 1.8-fold, respectively, that of untreated biomass. The pseudo-second-order model and the Freundlich isotherms described the adsorption processes better than the pseudo-first-order model and the Langmuir isotherms, respectively. The pre-treatments by surfactants increased the zeta potential and hydrophobicity of P. simplicissimum. Analysis of the cell surface by Fourier transform infrared spectrometry, energy dispersive X-ray, and environmental scanning electron microscopy indicated that the pre-treatments by surfactants changed the cell surface functional groups, element concentrations and micrographs. The results indicated that surfactants can be potentially used to increase phenol adsorption.

  15. Bioactive polyketides and alkaloids from Penicillium citrinum, a fungal endophyte isolated from Ocimum tenuiflorum.

    PubMed

    Lai, Daowan; Brötz-Oesterhelt, Heike; Müller, Werner E G; Wray, Victor; Proksch, Peter

    2013-12-01

    Chemical investigation of the endophytic fungus Penicillium citrinum cultured on white beans or on rice led to the isolation of two new alkaloids (1 and 2), along with fourteen known polyketides (6-12, 14-20) and four known alkaloids (3-5, and 13). The structures of the isolated compounds were determined by extensive analysis of the 1D, 2D NMR, and MS data, and by comparison with the literature. Compound 13, which had been previously obtained only by chemical synthesis, was isolated as a natural product for the first time, while compound 6 was firstly reported as a fungal metabolite. A re-isolation of sclerotinin A (14) revealed it to be a diastereoisomeric mixture (14a and 14b), whose stereochemistry was proposed for the first time based on ROESY experiment. All isolated compounds were evaluated for their cytotoxic and antibacterial activities. Compounds 12 and 17 showed significant cytotoxicity against the murine lymphoma cell line L5178Y with IC50 values of 1.0, and 0.78 μg/ml, respectively, while compounds 5, 11, and 15 were moderately active against Staphylococcus aureus ATCC 29213 (MIC 64 μg/ml).

  16. Cultivation conditions and properties of extracellular crude lipase from the psychrotrophic fungus Penicillium chrysogenum 9'.

    PubMed

    Bancerz, R; Ginalska, G; Fiedurek, J; Gromada, A

    2005-06-01

    Among 97 fungal strains isolated from soil collected in the arctic tundra (Spitsbergen), Penicillium chrysogenum 9' was found to be the best lipase producer. The maximum lipase activity was 68 units mL(-1) culture medium on the fifth day of incubation at pH 6.0 and 20 degrees C. Therefore, P. chrysogenum 9' was classified as a psychrotrophic microorganism. The non-specific extracellular lipase showed a maximum activity at 30 degrees C and pH 5.0 for natural oils or at pH 7.0 for synthetic substrates. Tributyrin was found to be the best substrate for lipase, among those tested. The Km and Vmax were calculated to be 2.33 mM and 22.1 units mL(-1), respectively, with tributyrin as substrate. The enzyme was inhibited more by EDTA than by phenylmethylsulfonyl fluoride and was reactivated by Ca2+. The P. chrysogenum 9' lipase was very stable in the presence of hexane and 1,4-dioxane at a concentration of 50%, whereas it was unstable in presence of xylene.

  17. Repeat induced point mutation in two asexual fungi, Aspergillus niger and Penicillium chrysogenum.

    PubMed

    Braumann, Ilka; van den Berg, Marco; Kempken, Frank

    2008-05-01

    Repeat induced point mutation (RIP) is a gene silencing mechanism present in fungal genomes. During RIP, duplicated sequences are efficiently and irreversibly mutated by transitions from C:G to T:A. For the first time, we have identified traces of RIP in transposable elements of Aspergillus niger and Penicillium chrysogenum, two biotechnologically relevant fungi. We found that RIP in P. chrysogenum has affected a large set of sequences, which also contain other mutations. On the other hand, RIP in A. niger is limited to only few sequences, but literally all mutations are RIP-like. Surprisingly, RIP occurred only in transposon sequences that have disrupted open reading frames in A. niger, a phenomenon not yet reported for other fungi. In both fungal species, we identified two sequences with strong sequence similarity to Neurospora crassa RID. RID is a putative DNA methyltransferase and the only known enzyme involved in the RIP process. Our findings suggest that both A. niger and P. chrysogenum either had a sexual past or have a sexual potential. These findings have important implications for future strain development of these fungi.

  18. More Than Just Oligomannose: An N-glycomic Comparison of Penicillium Species*

    PubMed Central

    Hykollari, Alba; Jin, Chunsheng; Yan, Shi; Vanbeselaere, Jorick; Razzazi-Fazeli, Ebrahim

    2016-01-01

    N-glycosylation is an essential set of post-translational modifications of proteins; in the case of filamentous fungi, N-glycans are present on a range of secreted and cell wall proteins. In this study, we have compared the glycans released by peptide/N-glycosidase F from proteolysed cell pellets of three Penicillium species (P. dierckxii, P. nordicum and P. verrucosum that all belong to the Eurotiomycetes). Although the major structures are all within the range Hex5–11HexNAc2 as shown by mass spectrometry, variations in reversed-phase chromatograms and MS/MS fragmentation patterns are indicative of differences in the actual structure. Hydrofluoric acid and mannosidase treatments revealed that the oligomannosidic glycans were not only in part modified with phosphoethanolamine residues and outer chain och1-dependent mannosylation, but that bisecting galactofuranose was present in a species-dependent manner. These data are the first to specifically show the modification of N-glycans in fungi with zwitterionic moieties. Furthermore, our results indicate that mere mass spectrometric screening is insufficient to reveal the subtly complex nature of N-glycosylation even within a single fungal genus. PMID:26515459

  19. Characterization and comparison of mitochondrial DNAs and rRNAs from Penicillium urticae and P. chrysogenum.

    PubMed

    Sekiguchi, J; Ohsaki, T; Yamamoto, H; Koichi, K; Shida, T

    1990-03-01

    Mitochondrial DNA (mt DNA) from a patulin producer, Penicillium urticae (synonym P. griseofulvum), was 27.8 kb +/- 0.6 kb in size by electron microscopy and 27.2 kb by agarose gel electrophoresis. Restriction endonuclease maps for nine restriction enzymes were constructed, and eleven fragments which covered the total range of the mt DNA were cloned into the Escherichia coli plasmid vector pUC19. Southern analysis of the native genomes of P. urticae and P. chrysogenum with six of the cloned fragments as probes indicated similar genome arrangements as well as similar restriction maps. Both the large and small rRNA genes of P. urticae and P. chrysogenum were located on these restriction maps using Southern hybridization, and the result also supported the similar arrangement. Agarose/formaldehyde gel electrophoresis indicated that the small rRNA was 1.5 kb in size in both species; but, surprisingly, the large rRNA was 4.2 kb in size for P. urticae and 3.5 kb for P. chrysogenum. These sizes were, respectively, 1.1 kb and 0.4 kb larger than those from the very closely related Aspergillus nidulans.

  20. Production and properties of alpha-amylase from Penicillium chrysogenum and its application in starch hydrolysis.

    PubMed

    Balkan, Bilal; Ertan, Figen

    2005-01-01

    Fungi were screened for their ability to produce alpha-amylase by a plate culture method. Penicillium chrysogenum showed high enzymatic activity. Alpha-amylase production by P. chrysogenum cultivated in liquid media containing maltose (2%) reached its maximum at 6-8 days, at 30 degrees C, with a level of 155 U ml(-1). Some general properties of the enzyme were investigated. The optimum reaction pH and temperature were 5.0 and 30-40 degrees C, respectively. The enzyme was stable at a pH range from 5.0-6.0 and at 30 degrees C for 20 min and the enzyme's 92.1% activity's was retained at 40 degrees C for 20 min without substrate. Hydrolysis products of the enzyme were maltose, unidefined oligosaccharides, and a trace amount of glucose. Alpha-amylase of P. chrysogenum hydrolysed starches from different sources. The best hydrolysis was determined (98.69%) in soluble starch for 15 minute at 30 degrees C.