Mycotoxins are toxic secondary metabolites that contaminate agricultural commodities and pose serious threats to human health. In this study, a multicopper oxidase, AfMCO, was identified and shown to efficiently degrade aflatoxin B1 (AFB1) and zearalenone (ZEN), with degradation rates of 99.5% and 98.7%, respectively. UPLC-Orbitrap-HRMS/MS analysis and zebrafish toxicity assays indicated that the degradation products exhibited significantly reduced toxicity. A triple mutant, M3 (M46H/T200P/E384I), obtained through computer-aided rational design based on AlphaFold3, FoldX, and PSSM, using ΔΔG < −2 kcal/mol and PSSM ≥ −1 as the mutant screening criteria, and exhibited a 55.05% increase in half-life at 50 °C, along with improved catalytic activity. Molecular dynamics simulations and homology modeling indicated that these mutations enhanced the overall structural stability of the enzyme. Moreover, M3 achieved degradation rates of 91.1% for AFB1 in peanut flour and 85.1% for ZEN in corn flour. These findings deepen the understanding of the molecular basis of AfMCO thermostability enhancement and support its application in the detoxification of contaminated feed.

Aflatoxin B1(AFB1), zearalenone(ZEN), and alternariol(AOH) are major mycotoxins that threaten food safety, yet enzymes capable of detoxifying these structurally diverse toxins remain limited. In this study, two enzymes from Bacillus velezensis HNGD-JQ06, laccase BVLac and superoxide dismutase BV2SOD, were identified as broad-spectrum detoxification candidates. BVLac degraded 93.33% of AFB1 at 60 °C and pH 7.0, 94.10% of ZEN at 60 °C and pH 8.0, and 96.87% of AOH at 50 °C and pH 8.0. BV2SOD degraded 92.59% of AFB1 and 91.99% of ZEN at 50 °C and pH 7.0, and 98.94% of AOH at 60 °C and pH 8.0. UHPLC-MS/MS analysis suggested that AFB1, ZEN, and AOH were transformed into products tentatively assigned as AFQ1/epi-AFQ1, 15-OH-ZEN, and a hydroxylated AOH derivative, respectively. Zebrafish assays confirmed reduced toxicity of the products, while molecular docking supported favorable enzyme-toxin binding. Both enzymes also retained activity in peanut, corn, and wheat flour matrices, highlighting their potential for multi-mycotoxin biodetoxification.

Aflatoxin B1 (AFB1) and Zearalenone (ZEN) are highly toxic mycotoxins. In this study, we combined FoldX-based computational mutation design with molecular-docking prescreening to systematically identify beneficial variants of the Bacillus-derived laccase. This workflow identified A225F as a promising mutation, increasing AFB1 detoxification activity by 32.8% within 1 h. Molecular dynamics simulations revealed that the phenylalanine substitution reshaped the internal microenvironment of the active site, compacting the substrate-binding cavity, and promoting more coordinated residue motions, collectively enhancing catalytic efficiency. UHPLC-MS/MS verified the conversion of AFB1 and ZEN into the less toxic products AFQ1 and 13-OH-ZEN-quinone, consistent with zebrafish hepatotoxicity assays. Under reaction conditions of 37 degrees C for 4 h, it efficiently and safely degraded more than 50% of AFB1 and ZEN in food matrices. These results demonstrate the effectiveness of a rational, computation-guided strategy for engineering CotA laccases with improved detoxification performance.

Metschnikowia pulcherrima includes strains of applied agro-food interest, particularly due to the antimicrobial activity against plant pathogens, contribution to the aroma of fermented beverages, and preliminary evidence related to probiotic activity. This biotechnological relevance sheds new light of interest on the biology of this yeast. To better understand and expand its biotechnological potential and applicability, the genomes of M. pulcherrima NRRL Y-7111 T, NRRL Y-48695, CBS 10357, and NRRL Y-48712 were sequenced, and de-novo assembled. Between 10,671 and 14,548 genes were predicted and the cooperative genomic analyses were integrated with experimental assessments relating to traits relevant for biotechnological application and safety. In silico and in vitro safety assessment revealed intermediate sensitivity for itraconazole; furthermore, variants of the genes related to pulcherrimin production and transport were found in all the genomes. Moreover, an arsenal of carbohydrate-active enzymes (CAZymes) was unravelled, and their predicted localization was investigated. This study expands the body of knowledge on M. pulcherrima, including traits relevant for defining its safety as a bioresource, which is a pivotal aspect for its possible inclusion in the European Food Safety Authority (EFSA) Qualified Presumption of Safety (QPS) list and its application in REgulated food/feed PROducts (REPRO) both in the European Union aligned European countries. • A pipeline for genomic characterisation and safety assessment of unconventional yeasts, using M. pulcherrima as a model species was developed. • M. pulcherrima strains can be considered safe and safety data can be used to develop a body of knowledge to include M. pulcherrima in EFSA QPS list. • Analysis of the predicted localization of CAZymes allowed the detection of compounds as potential biological control agents.

Laccases are promising enzymes for zearalenone (ZEN) degradation, especially when used with redox mediators in Laccase Mediator Systems (LMSs). Ery4 laccase and three mediators, 2,2 '-azino-bis (3-ethylbenzothiazoline-6sulfonic acid) (ABTS), 2,2,6,6-Tetramethylpiperidine-1-oxyl (TEMPO), and acetosyringone (AS), were tested for ZEN removal in vitro and in naturally contaminated corn. All mediators enabled complete ZEN degradation, with AS 1 mM and 1 U/mL Ery4 in only ten minutes. ZEN was reduced by 55.3 % in corn flour. By High-resolution mass spectrometry and Density Functional Theory (DFT) three major degradation products were identified: an oxidation product (14-hydroxyZENquinone), and for the first time, two AS- ZEN coupling products (AS-ZEN and AS-hydroxyZEN). In silico prediction revealed that 14-hydroxyZENquinone and AS-ZEN may possess lower estrogenic potential, indicating LMS as a safe detoxification strategy in food systems. These findings offer key insights into LMS for ZEN degradation and the underlying degradation pathway, thereby advancing its potential application in food safety strategies.

Pleurotus genus (Jacq.) P. Kumm comprises widely known edible mushrooms whose commercial and biotechnological exploitation has been steadily increasing worldwide. With the advent of modern DNA-based approach, the taxonomic definition of species within Pleurotus genus has undergone major changes but remains unclear. Furthermore, knowledge regarding the photobiology of Pleurotus and the role of light in regulating its primary and secondary metabolism, along with key commercial and biotechnological aspects, remains limited. This review aims to depict a comprehensive overview on Pleurotus genus, with a particular focus on its controversial taxonomy, biotechnological potential and photobiology and to provide significant insights to address future research on this topic and exploit light technology to maximize Pleurotus potential.

Fumonisin B1 (FB1) is the most harmful toxin, due to its incidence and high concentrations in maize, along with its toxicity to humans and animals. To investigate possible mechanisms of FB1 phytotoxicity, an RNA-Sequencing based transcriptome analysis was carried out at 3 h after FB1 treatment in the shoots of the CO433 maize line resistant to Fusarium verticillioides. One thousand four hundred and fifty-nine differentially expressed genes were identified, 13.9

Wild rocket (Diplotaxis tenuifolia (L.) DC cv. Dallas) is a leafy green vegetable appreciated for its pungent taste and healthy properties, often consumed as a ready-to-eat product. The cultivation system is crucial in determining the overall quality, while postharvest storage is fundamental for preserving nutritional quality, phytochemicals, and vitamins. This study aimed to investigate the phytochemical content and microbiological quality of soilless (SS) and soil-bound (SB) wild rocket during cold postharvest storage under blue, red, and green Light Emitting Diode (LED). Blue LED increased chlorophylls and carotenoids in SB after two days of storage, and chlorophyll a in SS after seven days. Furthermore, it reduced H2O2 levels after two days (SS and SB) and lipid peroxidation in SB. Red LED increased phenols in both SS and SB but was detrimental to chlorophyll, carotenoids, and oxidative markers. Green LED had less significant effects. Microbiological growth varied with LED treatment: green light increased mesophilic bacteria in SB, and red light did so in SS by day four, while blue light reduced bacterial growth at the end of storage. Overall, Blue LED was the most effective LED in preserving postharvest quality. Soilless cultivation was particularly beneficial in reducing lipid peroxidation and maintaining cell membrane integrity during long-term storage, and it might also be more effective in preserving ascorbic acid. Conversely, soil-bound cultivation methods could enhance initial polyphenol content or better preserve it during early storage. This study highlights the complex interplay of pre-harvest conditions, postharvest quality, and shelf-life performance.

Pistachio (Pistacia vera L.) is an economically important tree nut. Due to its nutritional properties and health benefits, it is considered a healthy food and thus widely consumed worldwide. However, fungal contamination of the commodities has received considerable attention because of possible contamination by toxigenic fungi, important source of mycotoxins, resulting from secondary metabolism and hazards to health consumer. Members of the genus Aspergillus, mainly Aspergillus flavus and Aspergillus niger, are reported as occurring most frequently on pistachio nuts, because able to grow in the presence of low amounts of water and to produce mycotoxins (aflatoxins and ochratoxins), that are well known for their harmful health effects on humans. Monitoring the contaminating fungal species is particularly worthy of note also in climate change scenario, allowing to notice changes in fungal population composition through the time. This study aimed to contribute to collect data about fungal population and mycotoxins occurred in pistachio samples collected in Turkey: prevalence of 2 species, A. flavus and Aspergillus tubingensis, was assessed. The A. flavus strains consisted of a mixed population of aflatoxin producers and non-producing strains in vitro, with evidence of a new genotype in gene cluster within strains of aflatoxin non-producing chemotype.

It has been widely assessed that the quality and quantity of nutrient supply and the growing system can strongly affect the growth and development of plants, the nutritional quality, and the levels of minerals, phytochemicals, and vitamins. The study was conducted on leaves of wild rockets grown in soil and soilless to examine the effect of growing conditions and mineral fertilization doses on the quality, mineral content, enzymatic and non-enzymatic antioxidants, and oxidative stress. The experiment was conducted in a plastic greenhouse into two independent sectors, one for soil-bound cultivation and another equipped for soilless cultivation. The crop was subjected to a high compared to a low-input fertilization program. Ion exchange chromatography, spectrophotometrically, and microbiological techniques were utilized. The soil-bound treatments increased the fresh weight, leaves number, chloride content, and microbial load. Exogenous application of higher nitrogen levels significantly boosted the ascorbate and hydrogen peroxide levels. Wild rocket growth in soilless showed a higher content of sulphates and polyphenols, and enhanced activity of the antioxidant enzymes dehydroascorbate reductase and monodehydroascorbate reductase. The fertilization rate and the cultivation system did not modify the content of nitrates, except in the soilless system treated with high fertilization program, where nitrate levels slightly exceeded regulatory limits. The yield and overall quality of wild rockets can be improved by combining the proper fertilizer dose with the growth system (soilless or soil) and suggested fertilization management is provided.

Aflatoxins (AFs) are toxic secondary metabolites produced by Aspergillus spp. and are found in food and feed as contaminants worldwide. Due to climate change, AFs occurrence is expected to increase also in western Europe. Therefore, to ensure food and feed safety, it is mandatory to develop green technologies for AFs reduction in contaminated matrices. With this regard, enzymatic degradation is an effective and environmentally friendly approach under mild operational conditions and with minor impact on the food and feed matrix. In this work, Ery4 laccase, acetosyringone, ascorbic acid, and dehydroascorbic acid were investigated in vitro, then applied in artificially contaminated corn for AFB1 reduction. AFB1 (0.1 µg/mL) was completely removed in vitro and reduced by 26% in corn. Several degradation products were detected in vitro by UHPLC-HRMS and likely corresponded to AFQ1, epi-AFQ1, AFB1-diol, or AFB1dialehyde, AFB2a, and AFM1. Protein content was not altered by the enzymatic treatment, while slightly higher levels of lipid peroxidation and H2O2 were detected. Although further studies are needed to improve AFB1 reduction and reduce the impact of this treatment in corn, the results of this study are promising and suggest that Ery4 laccase can be effectively applied for the reduction in AFB1 in corn.

A metabolic feature of lactic acid bacteria (LAB) is the production of exopolysaccharides (EPSs), which have technological and functional properties of interest to the food sector. The present study focused on the characterization of the Weissella cibaria strain C43-11, a high EPS producer in the presence of sucrose, in comparison with a low-producing strain (C2-32), and on possible genetic regulatory elements responsible for the modulation of dextransucrase (dsr) genes expression. NMR analysis of the polymeric material produced by the C43-11 strain indicated the presence of dextran consisting mainly of a linear scaffold formed by α-(1–6) glycosidic linkages and a smaller amounts of branches derived from α-(1–2), α-(1–3), and α-(1–4) linkages. Molecular analysis of the dsr genes and the putative transcriptional promoters of the two strains showed differences in their regulatory regions. Such variations may have a role in the modulation of dsr expression levels in the presence of sucrose. The strong upregulation of the dsr gene in the C43-11 strain resulted in a high accumulation of EPS. This is the first report showing differences in the regulatory elements of the dsr gene in W. cibaria and indicates a new perspective of investigation to identify the regulatory mechanism of EPS production.

We report the identification and characterisation of a mosaic, multidrug-resistant and mobilisable IncR plasmid (pST1023) detected in Salmonella ST1023, a monophasic variant 4,[5],12:i: strain of widespread pandemic lineage, reported as a Southern European clone. pST1023 contains exogenous DNA regions, principally gained from pSLT-derivatives and IncI1 plasmids. Acquisition from IncI1 included oriT and nikAB and these conferred the ability to be mobilisable in the presence of a helper plasmid, as we demonstrated with the conjugative plasmids pST1007-1D (IncFII) or pVC1035 (IncC). A sul3-associated class 1 integron, conferring resistance to aminoglycosides, chloramphenicol and trimethoprim-sulphonamides, was also embedded in the acquired IncI1 DNA segment. pST1023 also harboured an additional site-specific recombination system (rfsF/rsdB) and IS elements of the IS1, IS5 (IS903 group) and IS6 families. Four of the six IS26 elements present constituted two pseudo-compound-transposons, named PCT-sil and PCT-Tn10 (identified here for the first time). The study further highlighted the mosaic genetic architecture and the clinical importance of IncR plasmids. Moreover, it provides the first experimental data on the ability of IncR plasmids to be mobilised and their potential role in the horizontal spread of antimicrobial-resistant genes.

Lactic acid bacteria (LAB) decisively influence the technological, nutritional, organoleptic and preservation properties of bakery products. Therefore, their use has long been considered an excellent strategy to improve the characteristics of those goods. The aim of this study was the evaluation of microbial diversity in different doughs used for the production of a typical Apulian flatbread, named focaccia. Leavening of the analyzed doughs was obtained with baker’s yeast or by applying an innovative “yeast-free” protocol based on a liquid sourdough obtained by using Leuconostoc citreum strain C2.27 as a starter. The microbial populations of the doughs were studied by both a culture-dependent approach and metagenetic analyses. The flours used for dough preparation were also subjected to the same analyses. The metagenetic analyses were performed by sequencing the V5–V6 hypervariable regions of the 16S rRNA gene and the V9 hypervariable region of the 18S rRNA gene. The results indicate that these hypervariable regions were suitable for studying the microbiota of doughs, highlighting a significant difference between the microbial community of focaccia dough with baker’s yeast and that of the dough inoculated with the bacterial starter. In particular, the dough made with baker’s yeast contained a microbiota with a high abundance of Proteobacteria (82% of the bacterial population), known to be negatively correlated with the biochemical properties of the doughs, while the Proteobacteria in dough produced with the L. citreum starter were about 43.5% lower than those in flour and dough prepared using baker’s yeast. Moreover, the results show that the L. citreum C2.27 starter was able to dominate the microbial environment and also reveal the absence of the genus Saccharomyces in the dough used for the production of the “yeast-free” focaccia. This result is particularly important because it highlights the suitability of the starter strain for obtaining an innovative “yeast-free” product.

Red orange and lemon extract (RLE) is an anthocyanins-rich dietary supplement that may influence gastrointestinal bacterial community in ruminants. The aim of the present study was to investigate the RLE effects on gut microbiota composition in lambs. Twenty-eight lambs were randomly divided into a control group (CON; n = 14) and an anthocyanin group (ANT; n = 14) and fed the same diet; additionally, only the ANT received 90 mg/kg live weight of RLE at day. After lamb slaughter (40 ± 1 days), fecal samples were collected from the rectum and stored at −20 °C until analysis. Analysis of fecal microbiome was carried out by metabarcoding analysis of 16S rRNA. After reads denoising, sequences were aligned against SILVA rRNA sequence database using MALT, and taxonomic binning was performed with MEGAN. A significant increase in Firmicutes and Bacteroidetes and a decrease in Proteobacteria and Actinobacteria was observed in ANT compared to CON. Moreover, an interesting increase of Lactobacillus and Bifidobacterium genera and a decrease in Escherichia coli and Salmonella species were detected in ANT compared to CON. Results recommend that anthocyanin supplementation in lamb diet is able to modulate positively gut microbiota and may inhibit the growth of some potential pathogenic microorganisms.

Anaerobic digestion represents an interesting approach to produce biogas from organic waste materials contaminated by mycotoxins. In this study a shotgun metagenomic analysis of lab-scale bioreactors fed with mycotoxin-contaminated silage has been carried out to characterize the evolution of microbial community under the operating conditions and the key enzymatic activities responsible for mycotoxin degradation. The study was conducted at two different level of contamination for fumonisins and aflatoxin B1. After 15 days biogas production was not influenced by the presence of mycotoxins. Metagenomic analysis revealed that a high contamination rate of mycotoxins interfere with microbial diversity. Degradation of mycotoxins accounted in about 54% for aflatoxin B1 and 60% for fumonisins. The degradation activity of fumonisins resulted in the presence of partially hydrolyzed forms in both tested contamination levels. Accordingly, metagenomic functional analysis revealed the presence of two new carboxylesterase genes belonging to D. bacterium and P. bacterium putatively involved in fumonisin degradation. (C) 2020 The Author(s). Published by Elsevier Ltd.

Enzymatic catalysis is one of the main pillars of sustainability for industrial production. Enzyme application allows minimization of the use of toxic solvents and to valorize the agro-industrial residues through reuse. In addition, they are safe and energy efficient. Nonetheless, their use in biotechnological processes is still hindered by the cost, stability, and low rate of recycling and reuse. Among the many industrial enzymes, fungal laccases (LCs) are perfect candidates to serve as a biotechnological tool as they are outstanding, versatile catalytic oxidants, only requiring molecular oxygen to function. LCs are able to degrade phenolic components of lignin, allowing them to efficiently reuse the lignocellulosic biomass for the production of enzymes, bioactive compounds, or clean energy, while minimizing the use of chemicals. Therefore, this review aims to give an overview of fungal LC, a promising green and sustainable enzyme, its mechanism of action, advantages, disadvantages, and solutions for its use as a tool to reduce the environmental and economic impact of industrial processes with a particular insight on the reuse of agro-wastes.

Scientific communication is facilitated by a data-driven, scientifically sound taxonomy that considers the end-user's needs and established successful practice. In 2013, the Fusarium community voiced near unanimous support for a concept of Fusarium that represented a clade comprising all agriculturally and clinically important Fusarium species, including the F. solani species complex (FSSC). Subsequently, this concept was challenged in 2015 by one research group who proposed dividing the genus Fusarium into seven genera, including the FSSC described as members of the genus Neocosmospora, with subsequent justification in 2018 based on claims that the 2013 concept of Fusarium is polyphyletic. Here, we test this claim and provide a phylogeny based on exonic nucleotide sequences of 19 orthologous protein-coding genes that strongly support the monophyly of Fusarium including the FSSC. We reassert the practical and scientific argument in support of a genus Fusarium that includes the FSSC and several other basal lineages, consistent with the longstanding use of this name among plant pathologists, medical mycologists, quarantine officials, regulatory agencies, students, and researchers with a stake in its taxonomy. In recognition of this monophyly, 40 species described as genus Neocosmospora were recombined in genus Fusarium, and nine others were renamed Fusarium. Here the global Fusarium community voices strong support for the inclusion of the FSSC in Fusarium, as it remains the best scientific, nomenclatural, and practical taxonomic option available.

Aflatoxins (AFs) are secondary metabolites produced by Aspergillus spp., known for their hepatotoxic, carcinogenic, and mutagenic activity in humans and animals. AF contamination of staple food commodities is a global concern due to their toxicity and the economic losses they cause. Different strategies have been applied to reduce fungal contamination and AF production. Among them, the use of natural, plant-derived compounds is emerging as a promising strategy to be applied to control both Aspergillus spoilage and AF contamination in food and feed commodities in an integrated pre- and postharvest management. In particular, phenols, aldehydes, and terpenes extracted from medicinal plants, spices, or fruits have been studied in depth. They can be easily extracted, they are generally recognized as safe (GRAS), and they are food-grade and act through a wide variety of mechanisms. This review investigated the main compounds with antifungal and anti-aflatoxigenic activity, also elucidating their physiological role and the different modes of action and synergies. Plant bioactive compounds are shown to be effective in modulating Aspergillus spp. contamination and AF production both in vitro and in vivo. Therefore, their application in pre- and postharvest management could represent an important tool to control aflatoxigenic fungi and to reduce AF contamination.

Plant antioxidants are important compounds involved in plant defense, signaling, growth, and development. The quantity and quality of such compounds is genetically driven; nonetheless, light is one of the factors that strongly influence their synthesis and accumulation in plant tissues. Indeed, light quality affects the fitness of the plant, modulating its antioxidative profile, a key element to counteract the biotic and abiotic stresses. With this regard, light-emitting diodes (LEDs) are emerging as a powerful technology which allows the selection of specific wavelengths and intensities, and therefore the targeted accumulation of plant antioxidant compounds. Despite the unique advantages of such technology, LED application in the horticultural field is still at its early days and several aspects still need to be investigated. This review focused on the most recent outcomes of LED application to modulate the antioxidant compounds of plants, with particular regard to vitamin C, phenols, chlorophyll, carotenoids, and glucosinolates. Additionally, future challenges and opportunities in the use of LED technology in the growth and postharvest storage of fruits and vegetables were also addressed to give a comprehensive overview of the future applications and trends of research.