Sulfonylurea herbicides (SUHs) are frequently detected in aquatic environments and exhibit high persistence due to their structurally shielded reactive sites, posing potential ecological risks. Herein, a recyclable laccase-immobilized magnetic biochar system (L-MBC) was constructed for the efficient purification treatment of bensulfuron-methyl (BSM) and nicosulfuron (NIC). Compared with free laccase, L-MBC showed enhanced stability, environmental adaptability, and reusability. Under optimal conditions, removal efficiencies reached 88.7% for BSM and 80.0% for NIC within 48 h. L-MBC exhibited good matrix tolerance and retained nearly 50% removal of BSM and NIC after seven cycles. Based on this, L-MBC was combined with the fixed-bed continuous-flow column. Within 12 d, the device effectively treated 86.4 L of contaminated water. Mechanistic analysis indicated that L-MBC removed BSM/NIC through adsorption-enriched, mediator-assisted enzymatic oxidation, in which interfacial electron transfer promoted selective cleavage of the sulfonylurea bridge. Molecular docking and density functional theory (DFT) calculations confirmed the critical role of ABTS•+ in electron transfer. Transformation intermediates identified by UHPLC–HRMS indicated multiple pathways, including bond cleavage, hydroxylation, and de-substitution. Toxicity prediction (ECOSAR and T.E.S.T.), algal inhibition and soybean cultivation experiment showed that most intermediates exhibited lower ecological and health risks than the parent compounds. This study provides a concise and integrated strategy for the purification treatment and risk reduction of SUHs in complex aquatic environments.

Amasi, a traditional fermented milk produced in Southern Africa, is associated with several health benefits, such as probiotic activities, immune system modulation, and pharmacological (antimicrobial, antitumor and antioxidant) potential. This study investigated the microbial diversity in Amasi (produced from cow’s and goat’s milk) through targeted metagenomic bacterial 16S rRNA and fungal ITS sequencing, the metabolic functional prediction of Amasi samples using the Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) and profiled amino acids constituents using Liquid Chromatographic-Mass Spectrophotometry (LC-MS). The results obtained revealed Firmicutes, Bacteroidetes, and Proteobacteria as the most prevalent bacterial phyla, with Lactococcus and Lactobacillus being the most abundant genera. On the other hand, Ascomycota, Basidiomycota, and Mucoromycota were the main fungal phyla, while Aspergillus, Kazachstania, and Debaryomyces spp. dominated the fungal genera. Also, Pseudomonas spp., Bacillus spp., Clostridium spp., Cronobacter spp., Alternaria spp., Diaporthe spp., and Penicillium spp. were the probable pathogenic bacteria and fungi genera found, respectively. Atopobium, Synechococcus, and Parabacteroides were found less often as rare genera. It was found that the amino acid and drug metabolism pathway prediction values in Amasi samples were significantly higher (p < 0.05) than in raw cow and goat milk, according to the inferred analysis (PICRUSt). The amino acid validation revealed glutamine and asparagine values as the most significant (p < 0.05) for Amasi cow milk (ACM) and Amasi goat milk (AGM), respectively. Comparatively, ACM showed more microbial diversity than AGM, though there were relative similarities in their microbiome composition. PICRUSt analysis revealed significant metabolites in the two Amasi samples. Overall, data from this study showed heterogeneity in microbial diversity, abundance distributions, metabolites, and amino acid balance between raw cow/goat milk and Amasi samples.

Pesticide contamination in aquatic environments, particularly due to sulfonylurea herbicides, has become a growing concern. Herein, a highly porous biochar was synthesized using potassium bicarbonate (KHCO3) as a green activating agent through a one-step pyrolysis and acid washing process. Among the different samples, KBC-6 exhibited the highest specific surface area (3548.8 m2 g-1), the greatest defect density, and abundant oxygen-containing functional groups. As a result, KBC-6 demonstrated impressive adsorption capacities of 227.5 mg g-1 for bensulfuron-methyl (BSM) and 195.9 mg g-1 for nicosulfuron (NIC) at 298 K. The adsorption process followed pseudo-first-order kinetics and was well-described by the Freundlich model, with physical mechanisms, including pore filling, hydrogen bonding, and π-π interactions, contributing 49.4 % and 52.7 %, 15.7 % and 13.6 %, and 25.6 % and 23.9 % for BSM and NIC, respectively. KBC-6 achieved high removal efficiency of BSM and NIC across a wide range of conditions, including pH 5-9, 0.1 M anions, 10 M ionic strength, and 0-100 mg L-1 humic acid. After six regeneration cycles, KBC-6 maintained over 72.3 % removal efficiency. Fixed-bed column tests showed treatment volumes of 244.16 BV for BSM and 222.93 BV for NIC, and the treated effluent did not affect soybean root growth or fresh weight, indicating its safety. These results suggest that KHCO3-activated biochar provides a cost-effective and eco-friendly solution for removing sulfonylurea herbicides from polluted water.

The basidiomycete Crucibulum laeve strain LE-BIN1700 (Agaricales, Nidulariaceae) is able to grow on agar media supplemented with individual components of lignocellulose such as lignin, cellulose, xylan, xyloglucan, arabinoxylan, starch and pectin, and also to effectively destroy and digest birch, alder and pine sawdust. C. laeve produces a unique repertoire of proteins for the saccharification of the plant biomass, including predominantly oxidative enzymes such as laccases (family AA1_1 CAZymes), GMC oxidoreductases (family AA3_2 CAZymes), FAD-oligosaccharide oxidase (family AA7 CAZymes) and lytic polysaccharide monooxygenases (family LPMO X325), as well as accompanying acetyl esterases and loosenine-like expansins. Metabolomic analysis revealed that, specifically, monosaccharides and carboxylic acids were the key low molecular metabolites in the C. laeve culture liquids in the experimental conditions. The proportion of monosaccharides and polyols in the total pool of identified compounds increased on the sawdust-containing media. Multiple copies of the family AA1_1, AA3_2, AA7 and LPMOs CAZyme genes, as well as eight genes encoding proteins of the YvrE superfamily (COG3386), which includes sugar lactone lactonases, were predicted in the C. laeve genome. According to metabolic pathway analysis, the litter saprotroph C. laeve can catabolize D-gluconic and D-galacturonic acids, and possibly other aldonic acids, which seems to confer certain ecological advantages.

White rot fungi, especially representatives of the genus Trametes spp. (Polyporaceae), are effective destructors of various xenobiotics, including oestrogens (phenol-like steroids), which are now widespread in the environment and pose a serious threat to the health of humans, animals and aquatic organisms. In this work, the ability of the white rot fungus Trametes hirsuta LE-BIN 072 to transform oestrone (E1) and 17β-oestradiol (E2), the main endocrine disruptors, was shown. More than 90% of the initial E1 and E2 were removed by the fungus during the first 24 h of transformation. The transformation process proceeded predominantly in the direction of the initial substrates’ detoxification, with the radical oxidative coupling of E1 and E2 as well as their metabolites and the formation of less toxic dimers in various combinations. A number of minor metabolites, in particular, less toxic estriol (E3), were identified by HPLC-MS. The formation of E1 from E2 and vice versa were shown. The exoproteome of the white rot fungus during the transformation of oestrogens was studied in detail for the first time. The contribution of ligninolytic peroxidases (MnP5, MnP7 and VP2) to the process of the extracellular detoxification of oestrogens and their possible metabolites is highlighted. Thus, the studied strain appears to be a promising mycodetoxicant of phenol-like steroids in aquatic environments.

Recent consumer demand for non-dairy alternatives has forced many manufacturers to turn their attention to cereal-based non-alcoholic fermented products. In contrast to fermented dairy products, there is no defined and standardized starter culture for manufacturing cereal-based products. Since spontaneous fermentation is rarely suitable for large-scale commercial production, it is not surprising that manufacturers have started to adopt centuries-known dairy starters based on lactic acid bacteria (LABs) for the fermentation of cereals. However, little is known about the fermentation processes of cereals with these starters. In this study, we combined various analytical tools in order to understand how the most common starter cultures of LABs affect the most common types of cereals during fermentation. Specifically, 3% suspensions of rice, oat, and wheat flour were fermented by the pure cultures of 16 LAB strains belonging to five LAB species—Lacticaseibacillus paracasei, Lactobacillus delbrueckii, Lactobacillus helveticus, Streptococcus thermophilus, and Lactococcus lactis. The fermentation process was described in terms of culture growth and changes in the pH, reducing sugars, starch, free proteins, and free phenolic compounds. The organoleptic and rheological features of the obtained fermented products were characterized, and their functional properties, such as their antioxidant capacity and angiotensin-converting enzyme inhibitory activity, were determined.

Many different strains of Streptococcus thermophilus are commonly used as starter cultures, and search for new safe strains with desired industrial and probiotic properties is an important issue. In this article, genomes of two new S. thermophilus strains—Str16t and Str159—were sequenced, and the main genome characteristics were determined. In silico analysis of the sequenced genomes revealed the absence of transmissible antibiotic resistance genes, virulence genes associated with pathogenicity, and integrated plasmids; gene clusters encoding class I and class II bacteriocins were found. In vitro tests showed phosphatase, peptidase, β-galactosidase, and esterase activity of both strains. Both strains were capable of fermenting glucose, lactose, sucrose, and ribose; in addition, Str16t fermented mannose. In conclusion, it was demonstrated that both Str16t and Str159 are promising strains for application as starter and probiotic cultures.

Lignin peroxidase (LiP9) from the basidiomycete Trametes hirsuta LE-BIN 072, an effective lignin destructor, was purified to a homogeneous state (with an RZ purity index of 1.8) and characterized for the first time. The molecular weight of LiP9 was 43 kDa and its pI was 3.2. The enzyme showed the highest activity at pH 2.5 and 35°C when veratryl alcohol was used as a substrate. The analysis of the substrate specificity showed that LiP9 oxidized phenol derivatives much faster than those of benzoic and cinnamic acids with the same substituents in the benzene ring. The highest specific activity of the enzyme was observed for catechol oxidation. The ability of LiP9 to decolorize recalcitrant dyes (reactive black 5, congo red, remazol brilliant blue R, phenol red, indigo carmine, and bromocresol green) was assessed. The highest decolorization efficiency was shown for indigo carmine (in the presence of veratryl alcohol) and bromocresol green (directly) up to 80 and 60

Volatilomes of the basidial wood-destroying white rot fungus Trametes hirsuta LE-BIN 072 growing on a glucose–peptone medium (GP) with different types of wood sawdust (birch, alder, and pine) are represented by the following main groups of compounds: terpenes, aromatic compounds, fatty acid derivatives, and alkanes. The addition of sawdust to the cultivation medium led to a change in the spectrum of terpene and sesquiterpene compounds synthesized by fungus, as well as to a change in the ratio of the VOC component composition. On the control GP medium, β-elemene, β-barbatene, and γ-muurolene were detected; on the media with birch, alder, and pine sawdust, α-bulnesene, γ-cadinene, Δ3-carene, and camphene were detected. In silico analysis of the T. hirsuta 072 genome predicted ten genes encoding terpene synthases. Phylogenetic analysis showed that the proteins clustered with other basidiomycete terpene synthases into four major clades, suggesting that the enzymes in each cluster may produce related terpenes and sesquiterpenes.

As a toxic xenobiotic compound, the anthraquinone dye Remazol Brilliant Blue R (RBBR) poses a serious threat to aquatic ecosystems. In the present study, the ability of Trametes hirsuta to remove RBBR from the medium was investigated, and the role of adsorption by fungal mycelium and biodegradation by fungal enzymes was evaluated. It was shown that the whole fungal culture was able to remove up to 97% of the dye within the first four hours of incubation. Based on enzymatic activities in the culture broth, laccases were proposed to be the main enzymes contributing to RBBR degradation, and RT-qPCR measurements demonstrated an increase in transcription for the two laccase genes—lacA and lacB. Composite mycelial pellets of T. hirsuta with improved adsorption ability were prepared by adding activated carbon to the growth medium, and the induction of laccase activity by carbon was shown. For composite pellets, the RBBR decolorization degree was about 1.9 times higher at 1 h of incubation compared to carbon-free pellets. Hence, it was shown that using fungal mycelium pellets containing activated carbon can be an effective and economical method of dye removal.

Whole-genome sequencing was carried out, and the main characteristics of the genomes of three new strains of L. lactis AM1, MA1, and dlA, isolated from fermented milk and cereal grain products of South Africa and Russia, were established. The dual approach of in vitro and in silico analysis used in this work allowed us to assess more fully the bacteriocinogenic potential and antibiotic resistance of L. lactis strains. It was shown that L. lactis AM1 and dlA had an antagonistic effect against the test cultures of Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 6538. The growth retardation of E. coli and S. aureus after 48 hours of cocultivation was 65–75 and 30–50

Amasi, a traditional fermented milk produced in Southern Africa, is associated with several health benefits, such as probiotic activities, immune system modulation, antimicrobial, antitumor and antioxidant activity. In this study, microbial diversity in amasi (produced from cow and goat milk) was investigated by targeted metagenomic bacterial 16S rRNA and Fungal ITS sequencing. The results revealed Firmicutes, Bacteroidetes and Proteobacteria as the prevalent bacterial phyla, with Lactococcus being the prevalent bacterial genus. On the other hand, Ascomycota, Basidiomycota, Mucoromycota, and Rozellomycota were the main fungal phyla, while Aspergillus,Kazachstania, Debaryomyces, and Kluyveromyces spp. were the main fungal genera. Furthermore, of the bacteria genera identified, Bacillus (4%), Salmonella (0.85%), Escherichia-Shigella (0.38%), Staphylococcus (0.32%), Listeria (0.29%), Clostridium (0.28%), and Cronobacter (0.27%), could be considered pathogenic, while Atopobium, Synechococcus, and Parabacteroides occurred at lower frequencies as unique genera. Comparatively, the amasi produced from cow milk showed more microbial diversity than that produced from goat milk, though there were relative similarities in their microbiota composition. Functional prediction of drug and disease metabolism pathways revealed significant metabolites in the two amasi samples. Overall, data from this study showed heterogeneity in diversity and abundance distributions between raw cow/goat milk and amasi samples.

In this work, a comparative analysis of the profile of fatty acids (FA) and volatile organic compounds (VOCs) for skimmed cow’s milk fermented by four different probiotic strains of Lacticasibacillu paracasei (ABK, KF1, MA2, and MA3) was carried out. Analysis of FA and VOC profiles was performed using gas chromatography with mass spectrometric detection (GC-MS). For additional characterization of odor changes and its intensity in the fermented milk samples, the E-nose “electronic nose” odor analyzer was used. In total, the presence of 42 different FAs was detected in all samples, of which 17 were saturated, 8 monounsaturated, and 5 polyunsaturated. The strain-specific differences between the studied samples were of a complex nature and could not be explained only by the variation in the contribution of several individual FAs. The FA-nutritional indices characterizing the nutritional and biological value of the samples were different for milk fermented by different strains. Analysis of VOCs showed that the main odor-forming compounds in the studied samples were FAs and their reaction products, 2-heptanone, 2-nonanone, and 2-nonanol, which formed as a result of the following chain of transformations: β-oxidation → decarboxylation → reduction. The aroma of L. paracasei fermentation products, predicted on the basis of the odors of individual VOCs, generally coincided with their organoleptic assessment, a flavor typical of fermented milk (yogurt and curdled milk) with floral and fruity notes.

Being an abundant renewable source of aromatic compounds, lignin is an important component of future bio-based economy. Currently, biotechnological processing of lignin through low molecular weight compounds is one of the conceptually promising ways for its valorization. To obtain lignin fragments suitable for further inclusion into microbial metabolism, it is proposed to use a ligninolytic system of white-rot fungi, which mainly comprises laccases and peroxidases. However, laccase and peroxidase genes are almost always represented by many non-allelic copies that form multigene families within the genome of white-rot fungi, and the contributions of exact family members to the overall process of lignin degradation has not yet been determined. In this article, the response of the Trametes hirsuta LE-BIN 072 ligninolytic system to the presence of various monolignol-related phenolic compounds (veratryl alcohol, p-coumaric acid, vanillic acid, and syringic acid) in culture media was monitored at the level of gene transcription and protein secretion. By showing which isozymes contribute to the overall functioning of the ligninolytic system of the T. hirsuta LE-BIN 072, the data obtained in this study will greatly contribute to the possible application of this fungus and its ligninolytic enzymes in lignin depolymerization processes.

The interest in peroxidases of the basidiomycete secreted enzyme complex is due to their wide substrate specificity and the ability of these enzymes to participate in the biodegradation of such difficult to degrade biopolymers as lignin. However, due to the difficulty of isolating these enzymes from native sources, their study is difficult. In this work, expression vectors were created that carried the sequence encoding the T. hirsuta LE-BIN072 versatile peroxidase VP2, which was transformed into the genome of the P. canescens strain. Screening of transformants showed the presence of peroxidase activity up to 1 U/mL. Fragments of the target protein in the culture liquids of the selected transformants were identified by mass spectrometric analysis. A new strain, P. canescens pVP2D-6, a producer of the recombinant versatile peroxidase VP2 of T. hirsuta LE-BIN072, was obtained for the first time, and the ability of the enzyme complex secreted by it to modify alkaline lignin was shown.

Incorporation of probiotic Lacticaseibacillus paracasei into a standard yogurt starter culture can drastically improve its health promoting properties. However, besides being an advantage in itself, the incorporation of a new probiotic strain can significantly affect the overall composition of fermented milk. In this article, the effect of incorporation of the L. paracasei probiotic strains (KF1 and MA3) into several standard yogurt starter cultures (consisting of the following strains: Streptococcus thermophilus 16t and either Lactobacillus delbrueckii Lb100 or L. delbrueckii Lb200) was investigated. Such parameters as the degree of proteolysis, antioxidant activity, ACE-inhibitory activity, content of organic acids, profile of FAs and profile of volatile organic compounds were measured, and the influence of the starter culture composition on these parameters was described. It was demonstrated that, at least in the case of the studied strains, yogurt with L. paracasei had an advantage over the standard yogurt in terms of the content of acetoin, acetic acid, butyric acid and conjugated linoleic acid. Moreover, the incorporation of L. paracasei KF1 significantly improved the hypotensive properties of the resulting yogurt. Thus, the presented study provides insight into the bioactive molecules of probiotic yogurt and may be useful for both academia and industry in the development of new dairy-based functional products.

Basidiomycetes are attracting a lot of interest due to their remarkable ability to degrade phthalic acid esters (PAEs). In this article, the degradation process of benzyl butyl phthalate (BBP) and diisobutyl phthalate (DiBP) by the fungus Trametes hirsuta was studied. It was shown that T. hirsuta destroyed >95% of the initial BBP during 24 h, with complete destruction (>99%) occurring by the 3rd day of cultivation. DiBP was destroyed more slowly: 95% of the initial DiBP was destroyed by 10 days. The oxidase activity in the presence of PAEs increased by about 6.0 times, and the esterase activity by 2.0–2.5 times compared to the control. Enzymes involved in the process of PAEs degradation were identified on day 10: manganese peroxidases (MnP2, MnP5, MnP7), versatile peroxidase (VP2), lignin peroxidase (LiP9) and laccase (LacA). For the DiBP, the proportion of MnP5 and LiP9 increased, while the proportion of MnP2 and MnP7 decreased markedly. For the BBP, the proportion of MnP7 increased, while the proportion of VP2 significantly decreased. The GC–MS analysis of PAEs decomposition products were performed. For both PAEs, unique metabolites were found. A toxicity assessment revealed a pronounced toxic effect of metabolites formed during the degradation of BBP.

The ability of the white-rot fungus Peniophora lycii LE-BIN 2142 to degrade such phthalic acid esters (PAEs) as diethyl phthalate (DEP), dibutyl phthalate (DBP), di(2-ethylhexyl) phthalate (DEHP), diisobutyl phthalate (DiBP), and n-butylbenzyl phthalate (BBP) was studied. It was shown that DEHP was most efficiently biodegraded by the fungus (over 98