An Adams

JEM Spotlight: Fungi, mycotoxins and microbial volatile organic compounds in mouldy interiors from water-damaged buildings

Concerns have been raised about exposure to mycotoxin producing fungi and the microbial volatile organic compounds (MVOCs) they produce in indoor environments. Therefore, the presence of fungi and mycotoxins was investigated in 99 samples (air, dust, wallpaper, mycelium or silicone) collected in the mouldy interiors of seven water-damaged buildings. In addition, volatile organic compounds (VOCs) were sampled. The mycotoxins were analysed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) (20 target mycotoxins) and quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS). Morphological and molecular identifications of fungi were performed. Of the 99 samples analysed, the presence of one or more mycotoxins was shown in 62 samples by means of LC-MS/MS analysis. The mycotoxins found were mainly roquefortine C, chaetoglobosin A and sterigmatocystin but also roridin E, ochratoxin A, aflatoxin B(1) and aflatoxin B(2) were detected. Q-TOF-MS analysis elucidated the possible occurrence of another 42 different fungal metabolites. In general, the fungi identified matched well with the mycotoxins detected. The most common fungal species found were Penicillium chrysogenum, Aspergillus versicolor (group), Chaetomium spp. and Cladosporium spp. In addition, one hundred and seventeen (M)VOCs were identified, especially linear alkanes (C(9)-C(17)), aldehydes, aromatic compounds and monoterpenes.

Use of headspace SPME-GC-MS for the analysis of the volatiles produced by indoor molds grown on different substrates

An automated headspace solid phase microextraction method followed by GC-MS analysis was used to evaluate and compare the in vitro production of microbial volatile organic compounds (MVOCs) on malt extract agar, plasterboard and wallpaper. Five fungal strains were isolated from the walls of water-damaged houses and identified. In addition, four other common molds were studied. In general, MVOC production was the highest on malt extract agar. On this synthetic medium, molds typically produced 2-methylpropanol, 2-methylbutanol and 3-methylbutanol. On wallpaper, mainly 2-ethylhexanol, methyl 2-ethylhexanoate and compounds of the C8-complex such as 1-octene-3-ol, 3-octanone, 3-octanol and 1,3-octadiene were detected. The detection of 2-ethylhexanol and methyl 2-ethylhexanoate indicates an enhanced degradation of the substrate by most fungi. For growth on plasterboard, no typical metabolites were detected. Despite these metabolite differences on malt extract agar, wallpaper and plasterboard, some molds also produced specific compounds independently of the used substrate, such as trichodiene from Fusarium sporotrichioides and aristolochene from Penicillium roqueforti. Therefore, these metabolites can be used as markers for the identification and maybe also mycotoxin production of these molds. All five investigated Penicillium spp. in this study were able to produce two specific diterpenes, which were not produced by the other species studied. These two compounds, which remain unidentified until now, therefore seem specific for Penicillium spp. and are potentially interesting for the monitoring of this fungal genus. Further experiments will be performed with other Penicillium spp. to study the possibility that these two compounds are specific for this group of molds.

Biotransformation of (R)-(+)- and (S)-(−)-limonene to α-terpineol by Penicillium digitatum— investigation of the culture conditions

Abstract The biotransformation of (R)-(+)- and (S)-(−)-limonene by Penicillium digitatum was investigated. One strain of P. digitatum was able to convert (R)-(+)-limonene to pure (R)-(+)-α-terpineol in 8 h with a yield of up to 93%. It was found that (R)-(+)-limonene was converted much better into α-terpineol than (S)-(−)-limonene, and that no significant chemical conversion of the substrate occurred in control flasks at pH 3.5. The culture conditions involved such as the type and concentration of co-solvent applied and the sequential addition of substrate were investigated, taking into account some findings on the physical behaviour of the system. The highest bioconversion yields were obtained when the substrate was applied as a diluted solution in EtOH.

Formation of Pyrazines in Maillard Model Systems of Lysine-Containing Dipeptides

Whereas most studies concerning the Maillard reaction have focused on free amino acids, little information is available on the impact of peptides and proteins on this important reaction in food chemistry. Therefore, the formation of flavor compounds from the model reactions of glucose, methylglyoxal, or glyoxal with eight dipeptides with lysine at the N-terminus was studied in comparison with the corresponding free amino acids by means of stir bar sorptive extraction (SBSE) followed by GC-MS analysis. The reaction mixtures of the dipeptides containing glucose, methylglyoxal, and glyoxal produced 27, 18, and 2 different pyrazines, respectively. Generally, the pyrazines were produced more in the case of dipeptides as compared to free amino acids. For reactions with glucose and methylglyoxal, this difference was mainly caused by the large amounts of 2,5(6)-dimethylpyrazine and trimethylpyrazine produced from the reactions with dipeptides. For reactions with glyoxal, the difference in pyrazine production was rather small and mostly unsubstituted pyrazine was formed. A reaction mechanism for pyrazine formation from dipeptides was proposed and evaluated. This study clearly illustrates the capability of peptides to produce flavor compounds that can differ from those obtained from the corresponding reactions with free amino acids.

Mechanistic insights into furan formation in Maillard model systems.

Furan has recently received considerable attention as a possibly carcinogenic compound occurring in thermally processed foods. Although several food constituents have been identified as furan precursors, multiple formation pathways remain unclear. Therefore, the mechanisms of furan formation in Maillard model systems were studied by means of the carbon module labeling (CAMOLA) technique. Under both roasting and pressure-cooking conditions, furan was formed from glucose via the intact skeleton, and its formation pathways from glucose alone were not amino acid-dependent. However, some amino acids, especially alanine and serine, did influence the furan production by providing an additional formation pathway. Furthermore, most amino acids enhanced the furan production from glucose. Roasting conditions produced 25-100 times higher amounts of furan as compared to pressure-cooking conditions. Surprisingly, in the alanine/glucose model systems, the relative importance of furan production from glucose alone and from the combination of a glucose-derived and an alanine-derived fragment changed completely over a limited time course of 60 min.

Identification of volatile markers for indoor fungal growth and chemotaxonomic classification of Aspergillus species.

Microbial volatile organic compounds (MVOCs) were collected in water-damaged buildings to evaluate their use as possible indicators of indoor fungal growth. Fungal species isolated from contaminated buildings were screened for MVOC production on malt extract agar by means of headspace solid-phase microextraction followed by gas chromatography-mass spectrometry (GC-MS) analysis. Some sesquiterpenes, specifically derived from fungal growth, were detected in the sampled environments and the corresponding fungal producers were identified. Statistical analysis of the detected MVOC profiles allowed the identification of species-specific MVOCs or MVOC patterns for Aspergillus versicolor group, Aspergillus ustus, and Eurotium amstelodami. In addition, Chaetomium spp. and Epicoccum spp. were clearly differentiated by their volatile production from a group of 76 fungal strains belonging to different genera. These results are useful in the chemotaxonomic discrimination of fungal species, in aid to the classical morphological and molecular identification techniques.

Chemical modifications of peptides and their impact on food properties.

3.2. Peptide Backbone Modifications 7881 3.3. Peptide Side-Chain Modifications 7882 3.4. Peptide Cross-Linking 7883 3.5. Peptide Breakdown 7884 4. Reactions of Peptides with Carbonyl Compounds 7885 4.1. Reactions between Peptides and Carbohydrates 7885 4.1.

Influence of various growth parameters on fungal growth and volatile metabolite production by indoor molds.

A Penicillium polonicum, an Aspergillus ustus and a Periconia britannica strain were isolated from water-damaged environments and the production of microbial volatile organic compounds (MVOCs) was investigated by means of headspace solid-phase microextraction followed by GC-MS analysis. The most important MVOCs produced were 2-methylisoborneol, geosmin and daucane-type sesquiterpenes for P. polonicum, 1-octen-3-ol, 3-octanone, germacrene D, δ-cadinene and other sesquiterpenes for A. ustus and the volatile mycotoxin precursor aristolochene together with valencene, α-selinene and β-selinene for P. britannica. Different growth conditions (substrate, temperature, relative humidity) were selected, resembling indoor parameters, to investigate their influence on fungal metabolism in relation with the sick building syndrome and the results were compared with two other fungal strains previously analyzed under the same conditions. In general, the range of MVOCs and the emitted quantities were larger on malt extract agar than on wallpaper and plasterboard, but, overall, the main MVOC profile was conserved also on the two building materials tested. The influence of temperature and relative humidity on growth and metabolism is different for different fungal species, and two main patterns of behavior could be distinguished. Results show that, even at suboptimal conditions for growth, production of fungal volatiles can be significant.

Formation of pyrazines and a novel pyrrole in Maillard model systems of 1,3-dihydroxyacetone and 2-oxopropanal.

Alkylpyrazines are a very important class of Maillard flavor compounds, but their mechanism of formation is complex and consists of different pathways. The model reaction of 20 different amino acids with 1,3-dihydroxyacetone, as a precursor of 2-oxopropanal, was studied by means of SPME-GC-MS to investigate the involvement of the amino acid side chain in the substitution pattern of the resulting pyrazines. 2,5-Dimethylpyrazine was quantitatively the most important pyrazine formed from all of the amino acids. The amino acid side chain is not involved in its formation. The substituents of other less abundant pyrazines resulted mainly from the incorporation of the Strecker aldehyde or aldol condensation products in the intermediate dihydropyrazine. The importance of different reaction mechanisms was evaluated, taking into account the pattern of pyrazines identified. In the solvent extracts of aqueous model reactions of 2-oxopropanal with amino acids, the main reaction product was not a pyrazine but a novel pyrrole. This pyrrole was identified as 2,5-diacetyl-3-methyl-1 H-pyrrole by means of spectral analysis, secured by chemical synthesis. A reaction mechanism for its formation was proposed and evaluated. The influence of various reaction conditions on the formation of 2,5-diacetyl-3-methyl-1 H-pyrrole and 2,5-dimethylpyrazine in the model reaction of alanine with 2-oxopropanal was studied. These results underscore the importance of the ratio of the different reagents and the presence of water in the resulting flavor formation in the Maillard reaction.

Hypoglycemic and hypolipidemic effects of the total triterpene acid fraction from Folium Eriobotryae.

For seeking the good natural material to develop new agent to treat diabetes, the total triterpene acid (TTA) fraction extracted from Folium Eriobotryae [leaves of Eriobotrya japonica (Thunb.) Lindl.] was evaluated for its hypoglycemic and hypolipidemic potential through normal, alloxan and streptozotocin-induced diabetic mice administered with graded oral doses (100, 200, 300 mg/(kg day)) for 7 or 14 days. The results showed that a dose of 300 mg/kg of TTA is the most effective dose to cause significant (p<0.01) hypoglycemic and/or hypolipidemic effects on normal, alloxan and streptozotocin-induced diabetic mice. This dose also significantly (p<0.01) lowered the glycosylated serum protein (GSP), total cholesterol (TC) and triglyceride (TG) level in severely diabetic mice. Furthermore, TTA increased the superoxide dismutase activity (SOD) and the serum insulin level of diabetic mice. These evidences indicated that the total triperpene acid fraction from Folium Eriobotryae has a high anti-diabetic potential along with a good hypolipidemic profile.