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Scientific Reports May 2020The biodiversity and evolution of fungal communities were monitored over a period of 3 vintages in a new winery. Samples were collected before grape receipt and 3 months...
The biodiversity and evolution of fungal communities were monitored over a period of 3 vintages in a new winery. Samples were collected before grape receipt and 3 months after fermentation from 3 different wine related environments (WRE): floor, walls and equipment and analyzed using Illumina Mi-Seq. Genera of mold and filamentous fungi (294), non-enological (10) and wine-associated yeasts (25) were detected on all WREs before the arrival of the first harvest. Among them, genera like Alternaria and Aureobasidium persisted during two vintages. Therefore, these genera are not specific to winery environment and appear to be adapted to natural or anthropic environments due to their ubiquitous character. Some genera like Candida were also detected before the first harvest but only on one WREs, whereas, on the other WREs they were found after the harvest. The ubiquitous character and phenotypic traits of these fungal genera can explain their dynamics. After the first harvest and during 3 vintages the initial consortium was enriched by oenological genera like Starmerella introduced either by harvest or by potential transfers between the different WREs. However, these establishing genera, including Saccharomyces, do not appear to persist due to their low adaptation to the stressful conditions of winery environment.
Topics: Biodiversity; Biological Evolution; Fermentation; Fungi; Wine
PubMed: 32409784
DOI: 10.1038/s41598-020-64819-2 -
International Journal of Biological... May 2022Pullulan is a microbial polymer, commercially produced from Aureobasidium pullulans. Downstream processing of pullulan involves a multi-stage process which should be... (Review)
Review
Pullulan is a microbial polymer, commercially produced from Aureobasidium pullulans. Downstream processing of pullulan involves a multi-stage process which should be efficient, safe and reproducible. In liquid-liquid separations, firstly cell free extract is separated. Cell biomass can be separated after fermentation either by centrifugation or filtration. Due to practically insolubility of pullulan in organic solvents, ethanol and isopropanol are the most commonly used organic solvents for its recovery. Pullulan can also be purified by chromatographic techniques, but these are not cost effective for the purification of pullulan. Efficient aqueous two-phase system can be used for the purification of pullulan. The current review describes the methods and perspectives used for solid-liquid separation, liquid-liquid separations and finishing steps for the recovery of pullulan. Techniques used to determine the structural attributes of pullulan have also been highlighted.
Topics: Ascomycota; Fermentation; Glucans; Solvents
PubMed: 35354070
DOI: 10.1016/j.ijbiomac.2022.03.163 -
BMC Microbiology Aug 2020Yeasts, which are ubiquitous in agroecosystems, are known to degrade various xenobiotics. The aim of this study was to analyze the effect of fungicides on the abundance...
BACKGROUND
Yeasts, which are ubiquitous in agroecosystems, are known to degrade various xenobiotics. The aim of this study was to analyze the effect of fungicides on the abundance of natural yeast communities colonizing winter wheat leaves, to evaluate the sensitivity of yeast isolates to fungicides in vivo, and to select yeasts that degrade propiconazole.
RESULTS
Fungicides applied during the growing season generally did not affect the counts of endophytic yeasts colonizing wheat leaves. Propiconazole and a commercial mixture of flusilazole and carbendazim decreased the counts of epiphytic yeasts, but the size of the yeast community was restored after 10 days. Epoxiconazole and a commercial mixture of fluoxastrobin and prothioconazole clearly stimulated epiphyte growth. The predominant species isolated from leaves were Aureobasidium pullulans and Rhodotorula glutinis. In the disk diffusion test, 14 out of 75 yeast isolates were not sensitive to any of the tested fungicides. After 48 h of incubation in an aqueous solution of propiconazole, the Rhodotorula glutinis Rg 55 isolate degraded the fungicide in 75%. Isolates Rh. glutinis Rg 92 and Rg 55 minimized the phytotoxic effects of propiconazole under greenhouse conditions. The first isolate contributed to an increase in the dry matter content of wheat seedlings, whereas the other reduced the severity of chlorosis.
CONCLUSION
Not sensitivity of many yeast colonizing wheat leaves on the fungicides and the potential of isolate Rhodotorula glutinis Rg 55 to degrade of propiconazole was established. Yeast may partially eliminate the ecologically negative effect of fungicides.
Topics: Colony Count, Microbial; Fungicides, Industrial; Microbial Sensitivity Tests; Microbial Viability; Plant Leaves; Triazoles; Triticum; Yeasts
PubMed: 32758148
DOI: 10.1186/s12866-020-01885-6 -
Food Microbiology Dec 2022Processing, such as fresh cutting and drying, is essential to enhance profitability; therefore, to limit waste and reduce losses in fruit production such as mangoes....
Processing, such as fresh cutting and drying, is essential to enhance profitability; therefore, to limit waste and reduce losses in fruit production such as mangoes. Metabarcoding and microbial enumeration methods were utilized to explore the structure of mango microbiota, as well as their evolution after processing. Two mango ripening stages of cv. Cogshall were selected and processed into fresh-cut pieces or dried slices. Microbiological and physicochemical parameters were monitored during product storage, in order to assess the dynamics of quantitative and qualitative variations of the microbial flora. Proteobacteria was the dominant bacterial phylum of the mango surface and accounted for 73.16%, followed by Actinobacteria (10.16%), Bacteroidetes (7.82%) and Firmicutes (6.68%). Aureobasidium and Cladosporium were the only two genera shared between all types of samples (peel surface, dried slices and mango fresh-cut). However, the bacterial genera Lactobacillus and Pantoea were the most abundant in fresh-cut mango after 14 days of storage. Ascomycota was the dominant fungal phylum in the mango surface and accounted for 90.76% of the total number of detected sequences, followed by Basidiomycota (9.21%). In total, 866 microbial genera were associated with mango surface (562 bacterial and 304 fungal). Among detected yeast genera, Saccharomyces, Candida and Malassezia prevailed in mango flesh and were replaced by Wickerhamomyces after 14 days of storage. Alpha and beta diversity analyzes revealed differences in fungal and bacterial communities on fruit peel, in fresh-cut, dried slices, and during conservation (fresh-cut and dried slices). Mango processing (washing, peeling, cutting and drying) reduced the richness and the microbial diversity (bacterial and fungal) associated to the fruit, and drying limits the development of cultivable microorganisms during storage in comparison to fresh-cuts mangoes.
Topics: Bacteria; Food Handling; Mangifera; Mycobiome; Trees
PubMed: 36088111
DOI: 10.1016/j.fm.2022.104095 -
International Journal of Biological... Jan 2021Pullulan is an imperative microbial exo-polymer commercially produced by yeast like fungus Aureobasidium pullulans. Its structure contains maltosyl repeating units which... (Review)
Review
Pullulan is an imperative microbial exo-polymer commercially produced by yeast like fungus Aureobasidium pullulans. Its structure contains maltosyl repeating units which comprises two α-(1 → 4) linked glucopyranose rings attached to one glucopyranose ring through α-(1 → 6) glycosidic bond. The co-existence of α-(1 → 6) and α-(1 → 4) glycosidic linkages endows distinctive physico-chemical properties to pullulan. It is highly biocompatible, non-toxic and non-carcinogenic in nature. It is extremely resistant to any mutagenicity or immunogenicity. The unique properties of pullulan make it a potent candidate for biomedical applications viz. drug delivery, gene delivery, tissue engineering, molecular chaperon, plasma expander, vaccination, etc. This review highlights the potential of pullulan in biomedical research and development.
Topics: Animals; Blood Substitutes; Drug Carriers; Gene Transfer Techniques; Glucans; Humans; Tissue Scaffolds; Vaccination
PubMed: 33137388
DOI: 10.1016/j.ijbiomac.2020.10.227 -
Life (Basel, Switzerland) Dec 2023Consumers today seek safe functional foods with proven health-promoting properties. Current evidence shows that a healthy diet can effectively alleviate oxidative stress...
Consumers today seek safe functional foods with proven health-promoting properties. Current evidence shows that a healthy diet can effectively alleviate oxidative stress levels and reduce inflammatory markers, thereby preventing the occurrence of many types of cancer, hypertension, and cardiovascular and neurological pathologies. Nevertheless, as fruits and vegetables are mainly consumed fresh, they can serve as vectors for the transmission of pathogenic microorganisms associated with various disease outbreaks. As a result, there has been a surge in interest in the microbiome of fruits and vegetables. Therefore, given the growing interest in sweet cherries, and since their microbial communities have been largely ignored, the primary purpose of this study is to investigate their culturome at various maturity stages for the first time. A total of 55 microorganisms were isolated from sweet cherry fruit, comprising 23 bacteria and 32 fungi species. Subsequently, the selected isolates were molecularly identified by amplifying the 16S rRNA gene and ITS region. Furthermore, it was observed that the communities became more diverse as the fruit matured. The most abundant taxa included and among the bacteria, and , , and among the fungi.
PubMed: 38137924
DOI: 10.3390/life13122323 -
Journal of Alzheimer's Disease : JAD 2023Aureobasidium pullulans (black yeast) AFO-202 strain-produced beta glucan, Nichi Glucan, has been shown to improve the behavior and sleep pattern along with an increase... (Randomized Controlled Trial)
Randomized Controlled Trial
BACKGROUND
Aureobasidium pullulans (black yeast) AFO-202 strain-produced beta glucan, Nichi Glucan, has been shown to improve the behavior and sleep pattern along with an increase in α-synuclein and melatonin in children with autism spectrum disorder (ASD).
OBJECTIVE
In this randomized pilot clinical study, we have evaluated the gut microbiota of subjects with ASD after consumption of Nichi Glucan.
METHODS
Eighteen subjects with ASD were randomly allocated: six subjects in the control group (Group 1): conventional treatment comprising remedial behavioral therapies and L-carnosine 500 mg per day, and 12 subjects (Group 2) underwent supplementation with Nichi Glucan 0.5 g twice daily along with the conventional treatment for 90 days.
RESULTS
Whole genome metagenome (WGM) sequencing of the stool samples at baseline and after intervention showed that among genera of relevance, the abundance of Enterobacteriaceae was decreased almost to zero in Group 2 after intervention, whereas it increased from 0.36% to 0.85% in Group 1. The abundance of Bacteroides increased in Group 1, whereas it decreased in Group 2. The abundance of Prevotella increased while the abundance of Lactobacillus decreased in both Group 1 and Group 2. Among species, a decrease was seen in Escherichia coli, Akkermansia muciniphila CAG:154, Blautia spp., Coprobacillus sp., and Clostridium bolteae CAG:59, with an increase of Faecalibacterium prausnitzii and Prevotella copri, which are both beneficial.
CONCLUSION
AFO-202 beta 1,3-1,6 glucan, in addition to balancing the gut microbiome in children with ASD and its role in effective control of curli-producing Enterobacteriaceae that leads to α-synuclein misfolding and accumulation, may have a prophylactic role in Parkinson's and Alzheimer's diseases as well.
Topics: Humans; Gastrointestinal Microbiome; alpha-Synuclein; Glucans; Autism Spectrum Disorder; Neurodegenerative Diseases
PubMed: 36093695
DOI: 10.3233/JAD-220388 -
Carbohydrate Polymers Jul 2022Exopolysaccharide (EPS-1) was isolated from Aureobasidium pullulans CGMCC 23063 and purified with DEAE 650M and Sephadex G-100 column chromatography. The structural...
Exopolysaccharide (EPS-1) was isolated from Aureobasidium pullulans CGMCC 23063 and purified with DEAE 650M and Sephadex G-100 column chromatography. The structural characteristics and immunomodulatory activity of EPS-1 were investigated. The results showed EPS-1 (294.9 KDa) was β-glucan consisting of the backbone (1→3)-linked β-Glcp and β-(1,6)-branches by Ion chromatography, gas chromatography-mass spectrometry, and 1D/2D NMR spectroscopy. The SEM and AFM showed that EPS-1 composed spheres joined by triple-helix conformation into chains and circles. In vitro cell experiments, EPS-1 displayed immunological activity on the RAW264.7 cells. After being treated with EPS-1 (400 μg/mL), the concentration of NO, TNF-α, and IL-6 could reach 17.05 μM, 448.46 pg/mL, and 15.18 pg/mL, respectively. In addition, the cytokines mRNA levels also demonstrated the above results at the molecular level. The western-blot results demonstrated that the expression of phosphorylated p65 and p-erk associated with cytokine release, was increased with the dose of EPS-1.
Topics: Animals; Aureobasidium; Cytokines; Mice; Polysaccharides, Bacterial; RAW 264.7 Cells; beta-Glucans
PubMed: 35450628
DOI: 10.1016/j.carbpol.2022.119366 -
Journal of Fungi (Basel, Switzerland) Feb 2023Flax ( L.) is attacked by numerous devastating fungal pathogens, including , , and (). The effective control of flax diseases follows the paradigm of extensive...
Flax ( L.) is attacked by numerous devastating fungal pathogens, including , , and (). The effective control of flax diseases follows the paradigm of extensive molecular research on pathogenicity. However, such studies require quality genome sequences of the studied organisms. This article reports on the approaches to assembling a high-quality fungal genome from the Oxford Nanopore Technologies data. We sequenced the genomes of , , and () and received different volumes of sequencing data: 1.7 Gb, 3.9 Gb, and 11.1 Gb, respectively. To obtain the optimal genome sequences, we studied the effect of input data quality and genome coverage on assembly statistics and tested the performance of different assembling and polishing software. For , the most contiguous and complete assembly was obtained by the Flye assembler and the Homopolish polisher. The genome coverage had more effect than data quality on assembly statistics, likely due to the relatively low amount of sequencing data obtained for . The final assembly was 53.4 Mb long and 96.4% complete (according to the glomerellales_odb10 BUSCO dataset), consisted of 42 contigs, and had an N50 of 4.4 Mb. For and (), the best assemblies were produced by Canu-Medaka and Canu-Homopolish, respectively. The final assembly of had a length of 29.5 Mb, 99.4% completeness (dothideomycetes_odb10), an N50 of 2.4 Mb and consisted of 32 contigs. () assembly was 44.1 Mb long, 97.8% complete (hypocreales_odb10), consisted of 54 contigs, and had an N50 of 4.4 Mb. The obtained results can serve as a guideline for assembling a de novo genome of a fungus. In addition, our data can be used in genomic studies of fungal pathogens or plant-pathogen interactions and assist in the management of flax diseases.
PubMed: 36983469
DOI: 10.3390/jof9030301 -
Applied and Environmental Microbiology Apr 2021More than 30,000 tons of menthol are produced every year as a flavor and fragrance compound or as a medical component. So far, only extraction from plant material and...
More than 30,000 tons of menthol are produced every year as a flavor and fragrance compound or as a medical component. So far, only extraction from plant material and chemical synthesis are possible. An alternative approach for menthol production could be a biotechnological-chemical process with ideally only two conversion steps, starting from (+)-limonene, which is a side product of the citrus processing industry. The first step requires a limonene-3-hydroxylase (L3H) activity that specifically catalyzes hydroxylation of limonene at carbon atom 3. Several protein engineering strategies have already attempted to create limonene-3-hydroxylases from bacterial cytochrome P450 monooxygenases (CYPs, or P450s), which can be efficiently expressed in bacterial hosts. However, their regiospecificity is rather low compared to that of the highly selective L3H enzymes from the biosynthetic pathway for menthol in species. The only naturally occurring limonene-3-hydroxylase activity identified in microorganisms so far was reported for a strain of the black yeast-like fungus sp. in South Africa. We have discovered additional fungi that can catalyze the intended reaction and identified potential CYP-encoding genes within the genome sequence of one of the strains. Using heterologous gene expression and biotransformation experiments in yeasts, we were able to identify limonene-3-hydroxylases from and Further characterization of the enzyme demonstrated its high stereospecificity and regioselectivity, its potential for limonene-based menthol production, and its additional ability to convert α- and β-pinene to verbenol and pinocarveol, respectively. (-)-Menthol is an important flavor and fragrance compound and furthermore has medicinal uses. To realize a two-step synthesis starting from renewable (+)-limonene, a regioselective limonene-3-hydroxylase enzyme is necessary. We identified enzymes from two different fungi which catalyze this hydroxylation reaction and represent an important module for the development of a biotechnological process for (-)-menthol production from renewable (+)-limonene.
Topics: Ascomycota; Aureobasidium; Biotransformation; Catalysis; Cytochrome P-450 Enzyme System; Fungal Proteins; Hydroxylation; Industrial Microbiology; Limonene; Menthol
PubMed: 33637576
DOI: 10.1128/AEM.02873-20