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Item Chemical characterization and toxicity evaluation of fungal pigments for potential application in food, phamarceutical and agricultural industries(Elsevier B.V., 2023-03-31) Molelekoa, Tumisi Beiri Jeremiah; Augustyn, Wilma; Regnier, Thierry; Da Silva, Laura SuzanneConcerns associated with the use of synthetic colourants backs the demand for natural colourants. Thus, the current study aimed at characterizing crude fungal pigments produced by Penicillium multicolour, P. canescens, Talaromyces verruculosus, Fusarium solani and P. herquie. This included their antioxidant and antimicrobial properties together with acute toxicity evaluation on zebrafish embryos. The identification of pigment compounds was achieved through MS and IR data. The study demonstrated a substantial radical scavenging activity of extracts ranging from 65.49 to 74.46%, close to that of ascorbic acid (89.21%). Penicillium canescens and F. solani exhibited a strong antimicrobial activity against Escherichia coli and Enterococcus aerogenes and Salmonella typhi, Staphylococcus aureus and Bacillus cereus at MIC values ranging from 1.5 to 2.5 mg/mL. However, some levels of toxicity were observed for all extracts at a concentration range of 3–5 mg/mL. Pigment by P. multicolour, T. verruculosus and F. solani were tentatively identified through IR and MS data as sclerotiorin (yellow), rubropunctamine (red) and bostrycoidin (red). In conclusion, the study demonstrates a market potential of filamentous fungi pigments due to their antioxidant, antimicrobial activities, and prominent colours. Although there are some toxicity concerns, further tests must be done using molecular docking, albino mice and cell linings.Item Application and stability of fungal pigments using jelly sweets as a food model system.(Wiley, 2023-07-08) Molelekoa, Tumisi Beiri Jeremiah; Da Silva, Laura Suzanne; Regnier, Thierry; Augustyn, WilmaNatural pigments have seen an increase in usage over the past decades due to their acclaimed safety and active biological properties. This is in line with the shift in consumer market preferences requiring a source replacement for synthetic colourants. Filamentous fungi are amongst the natural sources currently explored for pigment production. In this study, the heat, pH and UV light stability of fungal pigments produced by Penicillium multicolour (dark brown), P. canescens (green) and P. herquie (yellow). Talaromyces verruculosus (red) and Fusarium solani (orange) were evaluated. The antioxidant properties of the pigments as influenced by these conditions were ascertained using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The pigments were also applied in a jelly sweet formulation and incubated at ambient temperature for 4 weeks. It was demonstrated that the pigment extracts of P. multicolour and T. verruculosus were significantly affected by higher pH (7.0 and 9.0), where the decline in antioxidant activity was observed. As far as high temperature and UV light exposure are concerned, all the pigment extracts, with exception of P. herquie, maintained an antioxidant activity above 35%. On the other hand, the L*a*b* values showed no overall significant change for heat and UV light exposure with the exception of P. canescens across the four pH levels (3.0, 4.0, 7.0 and 9.0), where an increase in the -a (green index) coupled with a decline in brightness (L* index) and b* (blue index) was observed. Finally, there was no significant change in colour over the 4 weeks of incubation. In conclusion, fungal pigments are generally stable at normal storage conditions (ambient temperature and normal light) and can be suitably applied in jelly sweets.Item Screening for volatile -unsaturated ester-producing yeasts from the feces of wild animals in South Africa.(MDPI, 2022-11-30) Tan, Mélissa; Caro, Yanis; Lebeau, Juliana; Shum-Cheong-Sing, Alain; François, Jean Marie; Regnier, Thierry; Petit, ThomasUnsaturated esters are fruity-aromatic compounds which are largely spread in the volatilome of many different fruits, but they are rarely found in the volatilome of yeasts. The yeast S. suaveolens has been recently shown to produce relatively high amounts of -unsaturated esters and it appears to be an interesting model for the production of these compounds. This study aimed to isolate new-unsaturated ester-producing yeasts by focusing on strains displaying a similar metabolism to S. suaveolens. While the production of -unsaturated esters by S. suaveolens is believed to be closely related to its ability to grow on media containing branched-chain amino acids (isoleucine, leucine and valine) as the sole carbon source (ILV+ phenotype), in this study, an original screening method was developed that selects for yeast strains displaying ILV+ phenotypes and is able to produce -unsaturated esters. Among the 119 yeast strains isolated from the feces of 42 different South African wild animal species, 43 isolates showed the ILV+ phenotype, among which 12 strains were able to produce -unsaturated esters. Two interesting - unsaturated esters were detected in two freshly isolated strains, both identified as Galactomyces candidus. These new esters were detected neither in the volatilome of the reference strain S. suaveolens, nor in any other yeast species previously studied for their aroma production. This work demonstrated the efficiency of an original method to rapidly screen for -unsaturated ester-producing yeasts. In addition, it demonstrated that wild animal feces are interesting resources to isolate novel strains producing compounds with original aromas.