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Selection of Non-Mycotoxigenic Inulinase Producers in the Group of Black Aspergilli for Use in Food Processing

Sanja Stojanović1orcid tiny, Jelena Stepanović2orcid tiny, Bojana Špirović Trifunović3orcid tiny, Nataša Duduk3orcid tiny, Biljana Dojnov1*orcid tiny, Bojan Duduk2orcid tiny and Zoran Vujčić4orcid tiny

1University of Belgrade, Institute of Chemistry, Technology and Metallurgy, National Institute of the Republic of Serbia, Department of Chemistry, Njegoševa 12, 1100 Belgrade, Republic of Serbia

2Institute of Pesticides and Environmental Protection, Banatska 31/b, 11080 Belgrade, Serbia

3University of Belgrade, Faculty of Agriculture, Nemanjina 6, 11080 Belgrade, Serbia

4University of Belgrade, Faculty of Chemistry, Department of Biochemistry, Studentski trg 12-16, 11000 Belgrade, Serbia

Article history:

Received: 7 October 2021

Accepted: 10 June 2022

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ochratoxin; fumonisin; Aspergillus spp.; fructooligosaccharides; inulinase


Research backgroundInulinases are used for fructooligosaccharide production and they are of interest for both scientific community and industry. Black aspergilli represent a diverse group of species that has use for enzyme production, in particular some species are known as potent inulinase producers. Finding new potential producers from the environment is as important as improving the production with known strains. Safe use of enzymes produced by aspergilli in food industry is placed ahead of their benefit for inulinase production.

Experimental approachHere we show a specific approach to finding/screening of newly isolated fungal inulinase producers that combines a newly developed screening method and an equally important assessment of the toxigenic potential of the fungus. In this study 39 black aspergilli collected from different substrates in Serbia were identified and assessed for inulinase production.

Results and conclusions. The most common species were Aspergillus tubingensis (51.2 %), followed by A. niger (23.1 %), A. welwitschiae (23.1 %) and A. uvarum (2.6 %). The isolates for inulinase production were selected using a cheap and easy, fast and non-hazardous alternative inulinase screening test developed in this work. Enzymatic activity of selected inulinase-producing strains was confirmed spectrophotometrically. Since some A. niger and A. welwitschiae strains are able to produce mycotoxins ochratoxin A (OTA) and fumonisins (FB), the toxigenic potential of selected inulinase producers was assessed analytically and genetically. Fungal enzyme producer can be considered safe for use in food industry only after comparing the results of both approaches for investigating toxic potential, the direct presence of mycotoxins in the enzyme preparation (analytically) and the presence of mycotoxin gene clusters (genetically). In some strains the absence of OTA and FB production capability was molecularly confirmed by the absence of complete or critical parts of biosynthetic gene clusters, respectively. The two best inulinase producers and mycotoxin non-producers (without mycotoxin production capability as additional safety) were selected as potential candidates for further development of enzyme production.

Novelty and scientific contributionThe presented innovative approach for the selection of potential fungal enzyme producer shows that only non-toxigenic fungi could be considered as useful in food industry. Although this study was done on local isolates, the approach is applicable globally.

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