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IUBMB Life Apr 2024Experimental evolution was carried out to investigate the adaptive responses of extremotolerant fungi to a stressful environment. For 12 cultivation cycles, the...
Experimental evolution was carried out to investigate the adaptive responses of extremotolerant fungi to a stressful environment. For 12 cultivation cycles, the halotolerant black yeasts Aureobasidium pullulans and Aureobasidium subglaciale were grown at high NaCl or glycerol concentrations, and the halophilic basidiomycete Wallemia ichthyophaga was grown close to its lower NaCl growth limit. All evolved Aureobasidium spp. accelerated their growth at low water activity. Whole genomes of the evolved strains were sequenced. No aneuploidies were detected in any of the genomes, contrary to previous studies on experimental evolution at high salinity with other species. However, several hundred single-nucleotide polymorphisms were identified compared with the genomes of the progenitor strains. Two functional groups of genes were overrepresented among the genes presumably affected by single-nucleotide polymorphisms: voltage-gated potassium channels in A. pullulans at high NaCl concentration, and hydrophobins in W. ichthyophaga at low NaCl concentration. Both groups of genes were previously associated with adaptation to high salinity. Finally, most evolved Aureobasidium spp. strains were found to have increased intracellular and decreased extracellular glycerol concentrations at high salinity, suggesting that the strains have optimised their management of glycerol, their most important compatible solute. Experimental evolution therefore not only confirmed the role of potassium transport, glycerol management, and cell wall in survival at low water activity, but also demonstrated that fungi from extreme environments can further improve their growth rates under constant extreme conditions in a relatively short time and without large scale genomic rearrangements.
PubMed: 38647201
DOI: 10.1002/iub.2825 -
International Journal of Food... Jun 2024Taggiasca table olives are typical of Liguria, a Northwestern Italian region, produced with a spontaneous fermentation carried out by placing the raw drupes directly...
Monitoring the yeasts ecology and volatiles profile throughout the spontaneous fermentation of Taggiasca cv. table olives through culture-dependent and independent methods.
Taggiasca table olives are typical of Liguria, a Northwestern Italian region, produced with a spontaneous fermentation carried out by placing the raw drupes directly into brine with a salt concentration of 8-12 % w/v. Such concentrations limit the development of unwanted microbes and favor the growth of yeasts. This process usually lasts up to 8 months. Yeasts are found throughout the entire fermentation process and they are mainly involved in the production of volatile organic compounds, which strongly impact the quality of the final product. The aim of this study was to evaluate the dynamics of autochthonous yeasts in brines and olives in a spontaneous process with no lye pre-treatment or addition of acids in the fermenting brine with 10 % NaCl (w/v) in two batches during 2021 harvest. Three hundred seventy-three yeast colonies were isolated, characterized by rep-PCR and identified by the D1/D2 region of the 26S rRNA gene sequencing. Mycobiota was also studied by 26S rRNA gene metataxonomics, while metabolome was assessed through GC-MS analysis. Traditional culture-dependent methods showed the dominance of Candida diddensiae, Wickerhamomyces anomalus, Pichia membranifaciens and Aureobasidium pullulans, with differences in species distribution between batches, sampling time and type of sample (olives/brines). Amplicon-based sequencing confirmed the dominance of W. anomalus in batch 1 throughout the entire fermentation, while Cyteromyces nyonsensis and Aureobasidium spp. were most abundant in the fermentation in batch 2. Volatilome results were analyzed and correlated to the mycobiota data, confirming differences between fermentation stages. Given the high appreciation for this traditional food, this study helps elucidate the mycobiota associated to Taggiasca cv. table olives and its relationship with the quality of the final product.
Topics: Fermentation; Olea; Volatile Organic Compounds; Yeasts; Food Microbiology; Italy; Salts
PubMed: 38615425
DOI: 10.1016/j.ijfoodmicro.2024.110688 -
Journal of Food Science May 2024Xylanases are mainly utilized in bakery industry for the hydrolysis of dietary fiber-based fractions. Their applications in gluten-free products have not been considered...
Exploitation of Aureobasidium pullulans NRRL Y-2311-1 xylanase in mulberry and rice flours-based gluten-free cookie formulation: Effects on dough properties and cookie characteristics.
Xylanases are mainly utilized in bakery industry for the hydrolysis of dietary fiber-based fractions. Their applications in gluten-free products have not been considered before. In the present study, the xylanase produced by Aureobasidium pullulans NRRL Y-2311-1 was utilized in a mulberry and rice flours-based gluten-free cookie formulation for the first time. Effects of various xylanase concentrations on gluten-free dough rheology and cookie characteristics were elucidated. Only rice flour-based cookie and only wheat flour-based cookie formulations were also prepared as comparison. Incorporation of xylanase into all cookie recipes resulted in softer cookie doughs with lower absolute stickiness. The hardness and absolute stickiness of the cookie doughs prepared by the mixture of mulberry and rice flours decreased by the addition of the enzyme into the formulation in a concentration-dependent manner. Enzyme concentrations above 100 U/100 g flour did not provide statistically significant further changes on gluten-free cookie doughs. Incorporation of xylanase into the cookie recipes resulted in increased baking loss and spread ratio in an enzyme concentration-dependent manner for all cookie types. Hardness values of both types of gluten-free cookies decreased by xylanase incorporation. Different effects on fracturability were observed depending on the cookie type and enzyme concentration. Enzyme concentration of 100 U/100 g flour provided mulberry and rice flours-based cookies with a more flexible and softer structure. No significant effects on color parameters of cookies were observed by xylanase incorporation.
Topics: Flour; Oryza; Morus; Diet, Gluten-Free; Rheology; Ascomycota; Food Handling; Endo-1,4-beta Xylanases; Hardness; Cooking; Dietary Fiber; Triticum; Glutens
PubMed: 38563094
DOI: 10.1111/1750-3841.17054 -
International Journal of Biological... Apr 2024Curcumin is a multitargeting nutraceutical with numerous health benefits, however, its efficacy is limited due to poor aqueous solubility and reduced bioavailability....
Curcumin is a multitargeting nutraceutical with numerous health benefits, however, its efficacy is limited due to poor aqueous solubility and reduced bioavailability. While nano-formulation has emerged as an alternative to encounter such issues, it often involves use of toxic solvents. Microbial synthesis may be an innovative solution to address this lacuna. Present study, for the first time, reports exploitation of Aureobasidium pullulans RBF4A3 for production of nano-curcumin. For this purpose, Aureobasidium pullulans RBF4A3 was inoculated in YPD media along with curcumin (0.1 mg/mL) and incubated for 24 h, 48 h, and 72 h. Subsequently, residual sugar, biomass, EPS concentration, curcumin concentration, and curcumin nanoparticle size were measured. As a result, nano-curcumin with an average particle size of 31.63 nm and enhanced aqueous solubility was obtained after 72 h. Further, investigations suggested that pullulan, a reducing polysaccharide, played a significant role in curcumin nano-formulation. Pullulan-mediated nano-curcumin formulation, with an average particle size of 24 nm was achieved with conversion rate of around 59.19 %, suggesting improved aqueous solubility. Additionally, the anti-oxidant assay of the resulting nano-curcumin was around 53.7 % per μg. Moreover, kinetics and thermodynamic studies of pullulan-based nano-curcumin revealed that it followed first-order kinetics and was favored by elevated temperature for efficient bio-conversion. Also, various physico-chemical investigations like FT-IR, NMR, and XRD reveal that pullulan backbone remains intact while forming curcumin nanoparticle. This study may open up new avenues for synthesizing nano-polyphenols through a completely green and solvent free process with plausible diverse applications.
Topics: Fermentation; Curcumin; Spectroscopy, Fourier Transform Infrared; Ascomycota; Water; Aureobasidium; Glucans
PubMed: 38521315
DOI: 10.1016/j.ijbiomac.2024.131088 -
Frontiers in Bioengineering and... 2024Polyol lipids (a.k.a. liamocins) produced by the polyextremotolerant, yeast-like fungus are amphiphilic molecules with high potential to serve as biosurfactants. So...
Polyol lipids (a.k.a. liamocins) produced by the polyextremotolerant, yeast-like fungus are amphiphilic molecules with high potential to serve as biosurfactants. So far, cultivations of have been performed in media with complex components, which complicates further process optimization due to their undefined composition. In this study, we developed and optimized a minimal medium, focusing on biosurfactant production. Firstly, we replaced yeast extract and peptone in the best-performing polyol lipid production medium to date with a vitamin solution, a trace-element solution, and a nitrogen source. We employed a design of experiments approach with a factor screening using a two-level-factorial design, followed by a central composite design. The polyol lipid titer was increased by 56% to 48 g L, and the space-time yield from 0.13 to 0.20 g L h in microtiter plate cultivations. This was followed by a successful transfer to a 1 L bioreactor, reaching a polyol lipid concentration of 41 g L. The final minimal medium allows the investigation of alternative carbon sources and the metabolic pathways involved, to pinpoint targets for genetic modifications. The results are discussed in the context of the industrial applicability of this robust and versatile fungus.
PubMed: 38511129
DOI: 10.3389/fbioe.2024.1379707 -
Molecular Biology of the Cell Apr 2024is a ubiquitous fungus with a wide variety of morphologies and growth modes including "typical" single-budding yeast, and interestingly, larger multinucleate yeast than...
is a ubiquitous fungus with a wide variety of morphologies and growth modes including "typical" single-budding yeast, and interestingly, larger multinucleate yeast than can make multiple buds in a single cell cycle. The study of promises to uncover novel cell biology, but currently tools are lacking to achieve this goal. Here, we describe initial components of a cell biology toolkit for , which is used to express and image fluorescent probes for nuclei as well as components of the cytoskeleton. These tools allowed live-cell imaging of the multinucleate and multibudding cycles, revealing highly synchronous mitoses in multinucleate yeast that occur in a semiopen manner with an intact but permeable nuclear envelope. These findings open the door to using this ubiquitous polyextremotolerant fungus as a model for evolutionary cell biology.
Topics: Saccharomyces cerevisiae; Ascomycota; Aureobasidium; Cytoskeleton
PubMed: 38446617
DOI: 10.1091/mbc.E23-10-0388 -
International Journal of Food... Apr 2024Alternaria alternata is a common fungal pathogen causing postharvest decay in table grapes. This study addressed the potential of autochthonous yeasts and bioactive...
Alternaria alternata is a common fungal pathogen causing postharvest decay in table grapes. This study addressed the potential of autochthonous yeasts and bioactive compounds of natural sources to act as biocontrol agents (BCAs) against A. alternata in cold-stored table grapes. With this purpose, 19 yeast capable of growing at 0-1 °C were isolated from the surface of Red Globe table grapes. These isolates, along with the pre-isolated strain Metschnikowia pulcherrima RCM2, were evaluated as BCAs in wounded berries. From these results, six yeast isolates were pre-selected to be combined with bioactive compounds of natural sources, like phenolic compounds (PCs) of side streams of wine industry, including bunch stem extract (BSE) (5-25 %), and cane extract (CE) (5-25 %), and functional polysaccharides from shrimp waste such as chitosan (CH) (0.5 %). Then, the biocontrol efficacy of combined treatments beyond individual ones was compared. The results revealed that 4 yeast isolates, namely M. pulcherrima RCM2 and ULA146, and Aureobasidium pullulans FUL14 and FUL18, were the most effective. However, when combined with the natural bioactive compounds, their efficacy against A. alternata did not increase significantly. Notably, ULA146 and FUL18 demonstrated a biocontrol efficacy of 36-37 %, comparable to that of the treatment with commercial doses of SO, which only showed a 27 % reduction in the lesion diameter. These findings highlight the potential of using psychrotrophic yeasts as BCAs against A. alternata in cold-stored table grapes. Combining these yeast strains with BSE, CE and CH did not increase BCAs efficacy against this pathogen at the concentrations tested. The development of effective biocontrol strategies for A. alternata could contribute to reducing reliance on chemically synthesized fungicides, promoting sustainable practices, aiming to improve the quality and safety of cold-stored table grapes.
Topics: Vitis; Yeasts; Alternaria; Fungicides, Industrial
PubMed: 38442539
DOI: 10.1016/j.ijfoodmicro.2024.110640 -
Journal of Bioscience and Bioengineering May 2024A high-yielding microbial polysaccharide-producing strain, named RM1603, was isolated from rhizosphere soil and identified by morphological and phylogenetic analysis....
A high-yielding microbial polysaccharide-producing strain, named RM1603, was isolated from rhizosphere soil and identified by morphological and phylogenetic analysis. The extracellular polysaccharides (EPS) were identified by thin-layer chromatography and infrared spectroscopy. The fermentation conditions were optimized by single factor experiments in shake flasks and a 5-L fermentor. The results of morphological and phylogenetic tree analysis showed that RM1603 was a strain of Aureobasidium pullulans. Its microbial polysaccharide was identified as pullulan, and the EPS production capacity reached 33.07 ± 1.03 g L in shake flasks. The fermentation conditions were optimized in a 5-L fermentor, and were found to encompass an initial pH of 6.5, aeration rate of 2 vvm, rotor speed of 600 rpm, and inoculum size of 2 %. Under these conditions, the pullulan yield of RM1603 reached 62.52 ± 0.24 g L. Thus, this study contributes RM1603 as a new isolation with high-yielding pullulan and potential application value in biotechnology.
Topics: Fermentation; Phylogeny; Ascomycota; Polysaccharides; Aureobasidium; Glucans
PubMed: 38413318
DOI: 10.1016/j.jbiosc.2023.12.018 -
Acta Myologica : Myopathies and... 2023Muscular dystrophies other than Duchenne muscular dystrophy (DMD) are genetic diseases characterized by increasing muscle weakness, loss of ambulation, and ultimately...
BACKGROUND
Muscular dystrophies other than Duchenne muscular dystrophy (DMD) are genetic diseases characterized by increasing muscle weakness, loss of ambulation, and ultimately cardiac and respiratory failure. There are currently no effective therapeutics available. Having demonstrated the efficacy of a N-163 strain of Aureobasidium Pullulans (Neu-REFIX) produced B-1, 3-1,6-Glucan in pre-clinical and clinical studies of Duchenne muscular dystrophy (DMD) earlier, we assessed the effectiveness of this novel Beta glucan in the other muscular dystrophies in the present study.
METHODS
In this 60-day study, six patients with muscular dystrophies other than DMD consumed one 8g gel of Neu-REFIX beta-glucan along with their usual standard of care treatment regimen, and their biomarkers of relevance to muscle function such as serum calcium (SC), creatine phosphokinase (CPK), and alkaline phosphatase (ALP) levels along with functional improvement criteria, which is, Medical research council (MRC) scale and North Star Ambulatory assessment (NSAA), assessed at baseline and following the intervention.
RESULTS
After the intervention, the SC levels significantly decreased from a mean baseline value of 9.28 mg/dL to 8.31 mg/dL (p-value = 0.02). With a p-value of 0.29, the mean CPK value dropped from 2192.33 IU/L to 1567.5 IU/L. Following the intervention, the ALP levels dropped from 200.33 to 75.5 U/L (p-value = 0.15). MRC scale improved in three out of six patients. NSAA remained stable. There were no adverse effects.
CONCLUSION
This study has proven the safety of Neu REFIX beta-glucan food supplement and its efficacy in improving both plasma biomarkers and functional parameters of muscle in a short duration of 2 months. Further validation by evaluation of muscle function for a longer duration is recommended to confirm the efficacy of Neu-REFIX food supplement as a potential adjuvant DMT in muscular dystrophies.
Topics: Humans; Muscular Dystrophy, Duchenne; beta-Glucans; Biomarkers; Muscles; Muscle Weakness
PubMed: 38406382
DOI: 10.36185/2532-1900-312 -
Microorganisms Feb 2024Post-harvest decay of fresh table grapes causes considerable annual production losses. The main fungal agents of decay both in pre- and post-harvest are , spp., spp.,...
Post-harvest decay of fresh table grapes causes considerable annual production losses. The main fungal agents of decay both in pre- and post-harvest are , spp., spp., spp., and spp. To date, the use of agrochemicals and SO are the main methods to control grape molds in pre- and postharvest, respectively. Significant improvements, however, have already been made in to apply innovative and more environmentally sustainable control strategies, such as Biological Control Agents (BCAs), which can reduce disease severity in both pre- and post-harvest. In this study, 31 new non- yeast strains, isolated from berries of native Apulian table grape genotypes, were tested for their in vivo effectiveness against grey mold of table grapes, resulting in two ('N22_I1' and 'S13_I3'), one ('N22_I3'), one ('OLB_9.1_VL') and one ('OLB_9.1_BR') yeast strains that were marked as efficient and good BCAs. Their mechanisms of action were characterized through in vitro assays, and additional characteristics were evaluated to assess the economic feasibility and viability for future technological employment. Their effectiveness was tested by reducing the working concentration, their antagonistic effect on a wide range of fungal pathogens, their ability to survive in formulations with long shelf life, and their safety to human health.
PubMed: 38399744
DOI: 10.3390/microorganisms12020340