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Biomedical subjects

Ulf Thrane

Publications and source records attributed to Ulf Thrane.

11 recordsLinked to original sources
PubMed2006

Colorimetric characterization for comparative analysis of fungal pigments and natural food colorants.

Exogenous pigments produced by ascomycetous filamentous fungi belonging to the genera Penicillium, Epicoccum, and Monascus, preselected based on chemotaxonomic knowledge, have been extracted and characterized by quantitative colorimetry. The color characteristics of the fungal extracts were compared to water soluble natural colorants derived from sources currently in use. The tested fungal extracts also included some commercially available Monascus colorants. The a values for the fungal extracts were found to be both positive and negative, the b values were found to be positive, while the hue angles of the fungal color extracts ranged from 40 to 110 indicating the color distribution of fungal extracts over the red-orange-yellow region of the CIELAB color space. The fungal extracts exhibited additional color hues in the red spectrum and similar hues in the yellow spectrum as compared to the reference natural colorants. They were also found to be similar or brighter in terms of chroma to some of the reference natural colorants. Principal component analysis was performed to group and distinguish different colors based on the a and b values. The fungal color extracts could be grouped in accordance with the similarity or difference in the color to those of the existing natural colorants. The diversity of colors was not only found among different fungal genera and/or species but also within the same species on changing the media. There was a marked change in the color composition of the extracts resulting in relatively different hues. Our results, thus, indicate that there exists pigment-producing genera of ascomycetous fungi other than Monascus that produce color shades in the red and the yellow spectra in addition or similar to reference colorants. These color shades could add to the color palette of the natural colorants currently in use. In addition, the multivariate approach in distinguishing and classifying the colorants was shown to be a very useful tool in colorimetric comparison of colorants.

Analysis of Variance↗
PubMed2006

The PKS4 gene of Fusarium graminearum is essential for zearalenone production.

Zearalenones are produced by several Fusarium species and can cause reproductive problems in animals. Some aurofusarin mutants of Fusarium pseudograminearum produce elevated levels of zearalenone (ZON), one of the estrogenic mycotoxins comprising the zearalenones. An analysis of transcripts from polyketide synthase genes identified in the Fusarium graminearum database was carried out for these mutants. PKS4 was the only gene with an enoyl reductase domain that had a higher level of transcription in the aurofusarin mutants than in the wild type. An Agrobacterium tumefaciens-mediated transformation protocol was used to replace the central part of the PKS4 gene with a hygB resistance gene through double homologous recombination in an F. graminearum strain producing a high level of ZON. PCR and Southern analysis of transformants were used to identify isolates with single insertional replacements of PKS4. High-performance liquid chromatography analysis showed that the PKS4 replacement mutant did not produce ZON. Thus, PKS4 encodes an enzyme required for the production of ZON in F. graminearum. Barley root infection studies revealed no alteration in the pathogenicity of the PKS4 mutant compared to the pathogenicity of the wild type. The expression of PKS13, which is located in the same cluster as PKS4, decreased dramatically in the mutant, while transcription of PKS4 was unchanged. This differential expression may indicate that ZON or its derivatives do not regulate expression of PKS4 and that the PKS4-encoded protein or its product stimulates expression of PKS13. Furthermore, both the lack of aurofusarin and ZON influenced the expression of other polyketide synthases, demonstrating that one polyketide can influence the expression of others.

Base Sequence↗
PubMed2005

Trichothecene production by Trichoderma brevicompactum.

Trichoderma brevicompactum, T. viride, T. harzianum, T. atroviride, T. longibrachiatum, T. erinaceum, T. citrinoviride, and Hypocrea lutea were screened for production of trichothecenes after growth on one or several solid and liquid media. Trichothecenes were detected by liquid chromatography combined with online UV/vis spectroscopy and electrospray high-resolution mass spectrometry. T. brevicompactum produced trichodermin and/or harzianum A on all media investigated, with liquid media yielding the largest amounts. Detection of octa-2Z,4E,6E-trienedioic acid in the harzianum-A-producing strains indicated that harzianum A was synthesized directly by esterification of trichodermol with octa-2Z,4E,6E-trienedioic acid. Both the T. viride strain from which trichodermin was originally isolated and the T. harzianum strain from which harzianum A was originally isolated were shown to belong to T. brevicompactum based on four independent criteria: metabolite profiles, micromorphology, macromorphology on yeast extract sucrose agar and potato dextrose agar, and DNA sequences of the ITS1/ITS2 regions of the nuclear ribosomal DNA.

Chromatography, Liquid↗
PubMed2005

Exploring fungal biodiversity for the production of water-soluble pigments as potential natural food colorants.

The production of many currently authorized natural food colorants has a number of disadvantages, including a dependence on the supply of raw materials and variations in pigment extraction. Fungi provide a readily available alternative source of naturally derived food colorants that could easily be produced in high yields. The recent authorization of a fungal food colorant has fuelled research to explore the extraordinary chemical diversity and biodiversity of fungi for the biotechnological production of pigments as natural food colorants. These studies require an appropriate use of chemotaxonomic tools and a priori knowledge of fungal metabolites to carry out intelligent screening for known or novel colorants as lead compounds. Such screening would result in the preselection of some potential pigment producers and the deselection of pathogenic strains and toxin producers. With advances in gene technology, in the future it should be possible to employ metabolic engineering to create microbial cell factories for the production of food colorants.

Biodiversity↗
PubMed2005

An oligonucleotide microarray for the identification and differentiation of trichothecene producing and non-producing Fusarium species occurring on cereal grain.

Cereal grain may be infected with a number of Fusarium species some of which are producers of highly toxic compounds such as the trichothecenes. Correct identification of these species is essential for risk assessment of cereal grain for human or animal consumption. Most of the available methods for identification are either time consuming or aimed at only one or a few target species. Microarray technology offers parallel analysis of a high number of DNA targets. In this study 57 capture oligonucleotides (CO) were designed based upon Fusarium ITS2 rDNA sequences, and used for microarray production. From this array COs could be selected that were able to hybridise specifically to labelled PCR products from the ITS region of Fusarium graminearum/Fusarium culmorum, Fusarium pseudograminearum, Fusarium poae, Fusarium sporotrichioides, Fusarium equiseti, Fusarium langsethiae and Fusarium tricinctum/Fusarium avenaceum. A few COs showed some cross hybridisation to non-target species. In a preliminary experiment it was shown that this cross hybridisation could be eliminated by increasing hybridisation stringency. The array could be used to detect individual Fusarium species in mixed samples and in environmental samples. This study demonstrates the feasibility of oligonucleotide microarrays for parallel detection of a number of Fusarium species.

DNA, Fungal↗
PubMed2005

Identification of cytotoxic principles from Fusarium avenaceum using bioassay-guided fractionation.

The cytotoxicity of extracts from rice cultures of five Fusarium avenaceum strains against the porcine epithelial kidney cell-line PK-15 was investigated using the Alamar Blue assay. After the identification of known fungal metabolites, cytotoxic extracts were fractionated using semi-preparative reversed-phase HPLC and normal phase LC, and the fractions were tested for cytotoxicity. In this way, two different groups of metabolites were identified as the major cytotoxic principles of the extracts. High concentrations of enniatins, especially enniatins B and B1, inhibited the metabolic activity of PK-15 cells. Furthermore, an unidentified metabolite, produced in high amounts by a strain that produced relatively small amounts of enniatins, was also found to be cytotoxic to PK-15 cells. This study shows that enniatins, a group of cyclic depsipeptides, which have been ignored as significant contributors to the toxicity of fungal extracts, may account for most of the observed effect for F. avenaceum.

Animals↗
PubMed2004

Diversity in metabolite production by Fusarium langsethiae, Fusarium poae, and Fusarium sporotrichioides.

The production of mycotoxins and other metabolites by 109 strains of Fusarium langsethiae, Fusarium poae, Fusarium sporotrichioides, and F. kyushuense was investigated independently in four laboratories by liquid or gas chromatography analyses of cultural extracts with UV diode array, electron capture, or mass spectrometric detection systems. From the compiled results, it was found that F. langsethiae consistently produced the trichothecenes diacetoxyscirpenol (DAS), T-2 toxin (T-2), HT-2 toxin (HT-2), and neosolaniol (NEO) and, to a lesser extent, some additional trichothecene derivatives. F. langsethiae also produced culmorins, chrysogine (CHRYS), aurofusarin (AUF), and enniatin (EN). F. sporotrichioides showed a metabolite profile similar to that of F. langsethiae, while F. poae had a different profile as 41 of 49 strains produced nivalenol (NIV) and other 8-keto trichothecenes, in addition to DAS and derivatives of this metabolite. Only a trace amount of NIV was detected from one strain of F. kyushuense. In summary, all the three core taxa of this joint study were found to produce trichothecenes. Fusarin C (F-C) was not detected from F. langsethiae, but it was produced by F. poae and F. sporotrichioides. Aurofusarin was only detected from a few strains of F. langsethiae, while nearly all strains of F. poae and F. sporotrichioides produced this compound. In contrast, chrysogine was not detected from F. poae, but was produced by the other two taxa. Production of enniatins was scattered among the three main taxa of this study, whereas beauvericin (BEA) was produced by many strains of F. poae and F. sporotrichioides. Only one odd strain of F. langsethiae (IBT 9959) produced beauvericin. However, the status of this strain is uncertain. By a polyphasic approach using species-specific metabolite profiles, the fruity odour of F. poae, and morphological observations, it was concluded that F. langsethiae, F. poae, and F. sporotrichioides should be regarded as three significant taxa at a species level.

Chromatography, Gas↗
PubMed2002

Production of trichothecenes and other secondary metabolites by Fusarium culmorum and Fusarium equiseti on common laboratory media and a soil organic matter agar: an ecological interpretation.

Fusarium culmorum and F. equiseti were characterized with regard to production of trichothecenes and other secondary metabolites. Results following growth on laboratory media are interpreted with the aim of increasing the understanding of fungal metabolism in the field environment. While trichothecene production was detected for 94 of 102 F. culmorum isolates, only 8 of 57 F. equiseti isolates were positive. Profiles of secondary metabolites were compared by following growth on yeast extract sucrose agar (YES), potato sucrose agar (PSA), and an agar medium, prepared from soil organic matter (SOM), which was included to simulate growth conditions in soil. SOM supported the production of chrysogine by F. culmorum. The two species utilized the media differently. F. culmorumproduced zearalenone (ZEA) on YES, whereas some F. equiseti isolates produced ZEA on PSA. Other F. equiseti isolates produced equisetin. These differences may reflect that F. culmorum depends on a pathogenic life style while F. equiseti has a more saprotrophic mode of existence.

Chromatography, High Pressure Liquid↗