Showing papers by "Ulf Thrane published in 2009"

Identification of potentially safe promising fungal cell factories for the production of polyketide natural food colorants using chemotaxonomic rationale

Abstract: Colorants derived from natural sources look set to overtake synthetic colorants in market value as manufacturers continue to meet the rising demand for clean label ingredients – particularly in food applications. Many ascomycetous fungi naturally synthesize and secrete pigments and thus provide readily available additional and/or alternative sources of natural colorants that are independent of agro-climatic conditions. With an appropriately selected fungus; using in particular chemotaxonomy as a guide, the fungal natural colorants could be produced in high yields by using the optimized cultivation technology. This approach could secure efficient production of pigments avoiding use of genetic manipulation. Polyketide pigment producing ascomycetous fungi were evaluated for their potential as production organisms based on a priori knowledge on species-specific pigment and potential mycotoxin production and BioSafety level (BSL) classification. Based on taxonomic knowledge, we pre-selected ascomycetous fungi belonging to Penicillium subgenus Biverticillium that produced yellow, orange or red pigments while deselecting Penicillium marneffei; a well known human pathogen in addition to other mycotoxigenic fungi belonging to the same group. We identified 10 strains belonging to 4 species; viz. P. purpurogenum, P. aculeatum, P. funiculosum, and P. pinophilum as potential pigment producers that produced Monascus-like pigments but no known mycotoxins. The selection/deselection protocol was illustrated in the pigment extracts of P. aculeatum IBT 14259 and P. crateriforme IBT 5015 analysed by HPLC-DAD-MS. In addition, extracellular pigment producing ability of some of the potential pigment producers was evaluated in liquid media with a solid support and N-glutarylmonascorubramine was discovered in the partially purified pigment extract of P. purpurogenum IBT 11181 and IBT 3645. The present work brought out that the use of chemotaxonomic tools and a priori knowledge of fungal extrolites is a rational approach towards selection of fungal polyketide pigment producers considering the enormous chemical diversity and biodiversity of ascomycetous fungi. This rationale could be very handy for the selection of potentially safe fungal cell factories not only for polyketide pigments but also for the other industrially important polyketides; the molecular and genetic basis for the biosynthesis of which has not yet been examined in detail. In addition, 4 out of the 10 chemotaxonomically selected promising Penicillium strains were shown to produce extracellular pigments in the liquid media using a solid support indicating future cell factory possibilities for polyketide natural food colorants.

Effect of temperature and water activity on the production of fumonisins by Aspergillus niger and different Fusarium species

Abstract: Fumonisins are economically important mycotoxins which until recently were considered to originate from only a few Fusarium species. However recently a putative fumonisin gene cluster was discovered in two different Aspergillus niger strains followed by detection of an actual fumonisin B2 (FB2) production in four strains of this biotechnologically important workhorse. In the present study, a screening of 5 A. niger strains and 25 assumed fumonisin producing Fusarium strains from 6 species, showed that all 5 A. niger strains produced FB2 and 23 of 25 Fusarium produced fumonisin B1 and other isoforms (fumonisin B2 and B3). Five A. niger and five Fusarium spp. were incubated at six different temperatures from 15-42°C on Czapek Yeast Agar +5% salt or Potato Dextrose Agar. A. niger had the highest production of FB2 at 25-30°C whereas Fusarium spp. had the maximal production of FB1 and FB2 at 20-25°C. Addition of 2.5-5% NaCl, or 10-20% sucrose increased the FB2 production of A. niger, whereas addition of glycerol reduced FB2 production. All three water activity lowering solutes reduced the fumonisin production of the Fusarium species. The present study shows that the regulation of fumonisin production is very different in A. niger and Fusarium, and that food and feeds preserved by addition of sugar or salts may be good substrates for fumonisin B2 production by A. niger.

Analysis of Fusarium avenaceum metabolites produced during wet apple core rot

Abstract: Wet apple core rot (wACR) is a well-known disease of susceptible apple cultivars such as Gloster, Jona Gold, and Fuji. Investigations in apple orchards in Slovenia identified Fusarium avenaceum, a known producer of several mycotoxins, as the predominant causal agent of this disease. A LC-MS/MS method was developed for the simultaneous detection of thirteen F. avenaceum metabolites including moniliformin, acuminatopyrone, chrysogine, chlamydosporol, antibiotic Y, 2-amino-14,16-dimethyloctadecan-3-ol (2-AOD-3-ol), aurofusarin, and enniatins A, A1, B, B1, B2, and B3 from artificially and naturally infected apples. Levels of moniliformin, antibiotic Y, aurofusarin, and enniatins A, A1, B, and B1 were quantitatively examined in artificially inoculated and naturally infected apples, whereas the remaining metabolites were qualitatively detected. Metabolite production was examined in artificially inoculated apples after 3, 7, 14, and 21 days of incubation. Most metabolites were detected after 3 or 7 days and reached significantly high levels within 14 or 21 days. The highest levels of moniliformin, antibiotic Y, aurofusarin, and the combined sum of enniatins A, A1, B, and B1 were 7.3, 5.7, 152, and 12.7 microg g(-1), respectively. Seventeen of twenty naturally infected apples with wACR symptoms contained one or more of the metabolites. Fourteen of these apples contained moniliformin, antibiotic Y, aurofusarin, and enniatins in levels up to 2.9, 51, 167, and 3.9 microg g(-1), respectively. Acuminatopyrone, chrysogine, chlamydosporol, and 2-AOD-3-ol were detected in 4, 11, 4, and 10 apples, respectively. During wet apple core rot, F. avenaceum produced high amounts of mycotoxins, which may pose a risk for consumers of apple or processed apple products.

Photostability of natural orange-red and yellow fungal pigments in liquid food model systems.

Abstract: The variation in the photostability among the currently authorized natural pigments limits their application span to a certain type of food system, and more robust alternatives are being sought after to overcome this problem. In the present study, the photostability of an orange-red and a yellow fungal pigment extract produced by ascomycetous fungi belonging to the genera Penicillium and Epicoccum , respectively, were studied in a soft drink model medium and in citrate buffer at low and neutral pH. The quantitative and qualitative color change pattern of the fungal pigment extracts indicated an enhanced photostability of fungal pigment extracts compared to the commercially available natural colorants Monascus Red and turmeric used as controls. Yellow components of the orange-red fungal pigment extract were more photostable than the red components. Chemistry of the photodegradation of the orange-red pigment extract was studied by high-performance liquid chromatography-diode array detection-mass spectrometry (HPLC-DAD-MS), and a light-induced formation of a structural analogue of sequoiamonascin C, a Monascus -like polyketide pigment discovered in the extract of Penicillium aculeatum IBT 14263 on yeast extract sucrose (YES) medium, was confirmed in the soft drink medium at pH 7.

Chemotaxonomic Exploration of Fungal Biodiversity for Polyketide Natural Food Colorants

Abstract: The variation in the photostability among the currently authorized natural pigments limits their application span to a certain type of food system and more robust alternatives are being sought after to overcome this problem. In the present study, the photostability of an orange-red and a yellow fungal pigment extract produced by ascomycetous fungi belonging to the genera Penicillium and Epicoccum, respectively, were studied in a soft drink model medium and in citrate buffer at low and neutral pH. The quantitative and qualitative color change pattern of the fungal pigment extracts indicated an enhanced photostability of fungal pigment extracts compared to the commercially available natural colorants Monascus Red and turmeric used as controls. Yellow components of the orange-red fungal pigment extract were more photostable than the red components. Chemistry of photodegradation of orange-red pigment extract was studied by HPLC-DAD-MS and a light induced formation of a structural analogue of sequoiamonascin C, a Monascus-like polyketide pigment discovered in the extract of Penicillium aculeatum IBT 14263 on YES medium, was confirmed in the soft drink medium at pH 7.