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Veterinary Clinical Pathology Jun 2024A 6-year-old 21.5 kg castrated male Siberian Husky was presented for acute onset of lethargy, vomiting, hemorrhagic diarrhea, and inappetence. Physical examination...
A 6-year-old 21.5 kg castrated male Siberian Husky was presented for acute onset of lethargy, vomiting, hemorrhagic diarrhea, and inappetence. Physical examination revealed marked discomfort upon abdominal palpation and 5%-7% dehydration. The CBC and biochemical profile revealed changes consistent with mild to moderate inflammation, dehydration, and gastrointestinal (GI) disease. Despite aggressive gastrointestinal support, anorexia persisted, and an upper GI endoscopy was performed in conjunction with esophagostomy tube placement. Endoscopy revealed abnormal gastric mucosa characterized by moderately well-demarcated areas of blue-black discoloration. Impression smears of a gastric biopsy revealed abundant extracellular yeasts with morphology most consistent with Candida spp. and frequent extracellular cocci. Similar yeast and bacteria, in lower numbers, were observed on cytologic analysis of a direct smear of the rectal mucosa. A rectal swab submitted for fungal culture yielded pure growth of fungal yeasts identified as Diutina (formerly Candida) rugosa by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. The dog's clinical signs improved with fluconazole, and he was discharged. Follow-up fungal culture of a rectal swab showed no growth of D. rugosa. To the authors' knowledge, this is the first case report that describes the clinical, hematologic, cytologic, and gross findings of enteric colonization by D. rugosa in a dog.
Topics: Animals; Male; Dogs; Dog Diseases; Candida; Candidiasis; Antifungal Agents; Fluconazole
PubMed: 38418373
DOI: 10.1111/vcp.13335 -
Bioorganic Chemistry Apr 2021Candida rugosa lipase (CRL) is an enzyme commonly used in medicinal and biotechnological applications. Allosteric modulators of CRL could aid in modifying lipase-related...
Candida rugosa lipase (CRL) is an enzyme commonly used in medicinal and biotechnological applications. Allosteric modulators of CRL could aid in modifying lipase-related diseases as well as improving biotechnological processes. Thus, a combinatorial approach of computational in-silico and high-throughput in-vitro screening was used to identify allosteric modulators of CRL. The screening of natural product libraries resulted in 132 compounds of which 53 were tested in-vitro. Subsequently, four inhibitors and three enhancers were identified of which rutin and cynaroside represented the strongest inhibitors of CRL activity (IC50: 227 ± 26 µM and 446 ± 15 µM, respectively) and NP-008496 the strongest enhancer (EC50: 425 ± 18 µM). All three compounds were predicted to bind the same allosteric site suggesting a common mechanism. Therefore, the present study demonstrated a reliable work-flow, identified an allosteric site of CRL and determined inhibitors and enhancers with numerous potential medical and biotechnological applications.
Topics: Allosteric Site; Biological Products; Dose-Response Relationship, Drug; Enzyme Inhibitors; Kinetics; Lipase; Molecular Docking Simulation; Molecular Structure; Saccharomycetales; Structure-Activity Relationship
PubMed: 33639364
DOI: 10.1016/j.bioorg.2021.104732 -
Journal of the Science of Food and... Apr 2021The biotechnological potential of yeasts from nuts such as pistachio, not only for health applications but also for industry use, has been scarcely studied. Interest in...
BACKGROUND
The biotechnological potential of yeasts from nuts such as pistachio, not only for health applications but also for industry use, has been scarcely studied. Interest in the probiotic capability of yeasts has increased in the past years as well as their utilization as food or feed preservatives. Their capabilities as biocontrol against problematic (spoilage or toxigenic) microorganisms or as antioxidants have been revalued. As a result, both abilities would be desirable to develop a new potential probiotic microorganism which could be added to food or feed to improve their properties.
RESULTS
Molecular techniques allowed the identification of a total of seven different species and 15 strains. A screening of the probiotic potential of these strains was carried out. It was found that 65% of the strains resisted the gastrointestinal conditions as well as presented a generation time of < 22 h. Additionally, some strains showed better kinetic parameters than Saccharomyces boulardii (positive control). Complementary tests were done to determine their auto-aggregation capacity, cell surface hydrophobicity, behaviour in a sequential simulated digestion, biofilm formation capability and carbon source assimilation. Finally, 67% and 13% of the studied yeasts showed biocontrol and antioxidant activities, respectively.
CONCLUSIONS
Diutina rugosa 14 followed by Diutina rugosa 8 were the best wild yeast from Pistacia vera as potential probiotic and in carbon source utilization. However, Hanseniaspora guilliermondii 6 and Aureobasidium proteae 5 could be used to improve food or feed product preservation because of their notable biocontrol and antioxidant capabilities. © 2020 Society of Chemical Industry.
Topics: Gastrointestinal Tract; Humans; Nuts; Phylogeny; Pistacia; Probiotics; Yeasts
PubMed: 32978783
DOI: 10.1002/jsfa.10839 -
Scientific Reports Jul 2020Microbial detoxification has been proposed as a new alternative for removing toxins and pollutants. In this study, the biodetoxification activities of yeasts against...
Microbial detoxification has been proposed as a new alternative for removing toxins and pollutants. In this study, the biodetoxification activities of yeasts against aflatoxin B and zinc were evaluated by HPLC and voltammetric techniques. The strains with the best activity were also subjected to complementary assays, namely biocontrol capability and heavy-metal resistance. The results indicate that the detoxification capability is toxin- and strain-dependent and is not directly related to cell growth. Therefore, we can assume that there are some other mechanisms involved in the process, which must be studied in the future. Only 33 of the 213 strains studied were capable of removing over 50% of aflatoxin B, Rhodotrorula mucilaginosa being the best-performing species detected. As for zinc, there were 39 strains that eliminated over 50% of the heavy metal, with Diutina rugosa showing the best results. Complementary experiments were carried out on the strains with the best detoxification activity. Biocontrol tests against mycotoxigenic moulds showed that almost 50% of strains had an inhibitory effect on growth. Additionally, 53% of the strains grew in the presence of 100 mg/L of zinc. It has been proven that yeasts can be useful tools for biodetoxification, although further experiments must be carried out in order to ascertain the mechanisms involved.
Topics: Aflatoxin B1; Biodegradation, Environmental; Chromatography, High Pressure Liquid; Environmental Pollutants; Food Safety; Metals, Heavy; Pichia; Rhodotorula; Saccharomyces cerevisiae; Saccharomycetales; Wastewater; Yeasts; Zinc
PubMed: 32647290
DOI: 10.1038/s41598-020-68154-4 -
Applied Biochemistry and Biotechnology Feb 2021The potential use of alternative culture media towards the development of a sustainable bioprocess to produce lipases by Diutina rugosa is clearly demonstrated. First, a...
The potential use of alternative culture media towards the development of a sustainable bioprocess to produce lipases by Diutina rugosa is clearly demonstrated. First, a synthetic medium containing glucose, peptone, yeast extract, oleic acid, and ammonium sulfate was proposed, with lipase activity of 143 U/L. Then, alternative culture media formulated with agro-industrial residues, such as molasses, corn steep liquor (CSL), and olive mill waste (OMW), were investigated. An experimental design was conducted, and only CSL concentration was found to have a positive effect in lipase production. The highest lipase activity (561 U/L) was produced on a mixture of molasses (5 g/L), CSL (6 g/L), OMW (0.5% v/v), 0.5 g/L of ammonium sulfate, and 3 g/L of peptone at 24 h of cultivation. Lipase production was also carried out in a 1-L bioreactor leading to a slightly higher lipase activity at 24 h of cultivation. The semi-purified enzyme exhibits an optimum temperature and pH of 40 °C and 7.0, respectively. Finally, the media cost per unit of lipase produced (UPC) was influenced by the medium components, specially by the inducer used. The lowest UPC was obtained when the agro-industrial residues were combined and used at the improved concentrations.
Topics: Bioreactors; Culture Media; Fungal Proteins; Industrial Microbiology; Lipase; Saccharomycetales; Waste Disposal, Fluid
PubMed: 33043398
DOI: 10.1007/s12010-020-03431-6 -
Journal of Chemical Information and... Feb 2021Lipases are enzymes able to catalyze the hydrolysis or synthesis of triglycerides, depending on the reaction conditions, whereas sterol esterases show the same ability...
Lipases are enzymes able to catalyze the hydrolysis or synthesis of triglycerides, depending on the reaction conditions, whereas sterol esterases show the same ability on sterol esters. Structurally, both kinds of enzymes display an α/β-hydrolase fold, with a substrate-binding pocket formed by a hydrophobic cavity covered by a mobile lid. However, it has been reported that some lipases from the -like family display wide substrate specificity on both triglycerides and sterol esters. Among them, enzymes with different biotechnological applications, such as the lipase isoenzymes produced by and the sterol esterase from , have been exhaustively characterized and their crystal structures are available. Differences in substrate affinity among these proteins have been attributed to changes in their hydrophobicity. In this work, we analyzed the full catalytic mechanisms of these proteins using molecular dynamics tools, gaining insight into their mechanistic properties. In addition, we developed an protocol to predict the substrate specificity using and lipases as model enzymes and triglycerides and cholesterol esters with different fatty acid chain lengths as model substrates. The protocol was validated by comparing the results with those described in the literature. These results would be useful to perform virtual screening of substrates for enzymes of the -like family with unknown catalytic properties.
Topics: Candida; Lipase; Ophiostoma; Saccharomycetales; Sterol Esterase; Substrate Specificity
PubMed: 33555857
DOI: 10.1021/acs.jcim.0c01151 -
Bioengineering (Basel, Switzerland) Aug 2023This work demonstrated the feasibility of an industrial-scale aerated static pile composting system for treating one of the common biowastes-soybean curd residue. The...
This work demonstrated the feasibility of an industrial-scale aerated static pile composting system for treating one of the common biowastes-soybean curd residue. The mixing ratios of the feedstock were optimized to achieve a carbon-nitrogen ratio and a moisture level in the ranges of 25-35 and 60-70%, respectively. This open-air composting system required 6-7 months to obtain a mature compost. Solvita and seed germination tests further confirmed the maturity of the compost, with 25% compost extract concentration yielding the best germination index in the absence of phytotoxicity. The bacterial and fungal compositions of the compost piles were further examined with metagenomic analysis. spp., spp., and spp. were among the unique bacteria found, and , , and were among the unique fungi found in the compost piles, suggesting the presence of good microorganisms for degrading the organic biowastes.
PubMed: 37627823
DOI: 10.3390/bioengineering10080938 -
Biomolecules Jun 2021Water content is an important factor in lipase-catalyzed reactions in organic media but is frequently ignored in the study of lipases by molecular dynamics (MD)...
Water content is an important factor in lipase-catalyzed reactions in organic media but is frequently ignored in the study of lipases by molecular dynamics (MD) simulation. In this study, lipase B, lipase and lipase were used as research models to explore the mechanisms of lipase in micro-aqueous organic solvent (MAOS) media. MD simulations indicated that lipases in MAOS systems showed unique conformations distinguished from those seen in non-aqueous organic solvent systems. The position of water molecules aggregated on the protein surface in MAOS media is the major determinant of the unique conformations of lipases and particularly impacts the distribution of hydrophilic and hydrophobic amino acids on the lipase surface. Additionally, two maxima were observed in the water-lipase radial distribution function in MAOS systems, implying the formation of two water shells around lipase in these systems. The energy landscapes of lipases along solvent accessible areas of catalytic residues and the minimum energy path indicated the dynamic open states of lipases in MAOS systems differ from those in other solvent environments. This study confirmed the necessity of considering the influence of the microenvironment on MD simulations of lipase-catalyzed reactions in organic media.
Topics: Basidiomycota; Cellular Microenvironment; Computational Biology; Fungal Proteins; Hydrophobic and Hydrophilic Interactions; Lipase; Molecular Dynamics Simulation; Protein Structure, Secondary; Protein Structure, Tertiary; Saccharomycetales; Water
PubMed: 34200257
DOI: 10.3390/biom11060848 -
Angewandte Chemie (International Ed. in... Nov 2020Enzyme-powered micro/nanomotors have myriads of potential applications in various areas. To efficiently reach those applications, it is necessary and critical to...
Enzyme-powered micro/nanomotors have myriads of potential applications in various areas. To efficiently reach those applications, it is necessary and critical to understand the fundamental aspects affecting the motion dynamics. Herein, we explored the impact of enzyme orientation on the performance of lipase-powered nanomotors by tuning the lipase immobilization strategies. The influence of the lipase orientation and lid conformation on substrate binding and catalysis was analyzed using molecular dynamics simulations. Besides, the motion performance indicates that the hydrophobic binding (via OTES) represents the best orienting strategy, providing 48.4 % and 95.4 % increase in diffusion coefficient compared to hydrophilic binding (via APTES) and Brownian motion (no fuel), respectively (with C of 100 mm). This work provides vital evidence for the importance of immobilization strategy and corresponding enzyme orientation for the catalytic activity and in turn, the motion performance of nanomotors, and is thus helpful to future applications.
Topics: Hydrophobic and Hydrophilic Interactions; Lipase; Molecular Dynamics Simulation; Nanotechnology; Particle Size; Protein Conformation; Saccharomycetales; Surface Properties
PubMed: 32755070
DOI: 10.1002/anie.202008339 -
Journal of Applied Microbiology Mar 2021This study aims to identify the yeast species and strains which entitled an unstudied area of Spain and evaluate the yeast species diversity richness and the genetic...
AIMS
This study aims to identify the yeast species and strains which entitled an unstudied area of Spain and evaluate the yeast species diversity richness and the genetic variety.
METHODS AND RESULTS
A total of 702 yeasts were isolated from different environments in a central Spanish region (La Mancha) with diverse sources of origin (food, animals, flowers and environmental sources) during spring season. Thanks to the analysis carried out by the PCR-RFLP technique and sequencing, 35 species were identified. A neighbour-joining phylogenetic tree was created based on D1/D2 sequences. Moreover 330 strains were determined by PCR-RAPD and their profiles were analysed using the bioinformatics programme BioNumerics 7·6. The Simpson's index (D) and the genetic diversity percentage were calculated with the aim of studying the richness of the species in each environment and the genetic variety in each species.
CONCLUSIONS
This study has permitted to know that the majority of the species found was Diutina rugosa while the most ubiquitous was Rhodotorula mucilaginosa which expose the dispersion capability of this species. The diversity parameters has revealed that the highest species richness was associated to environmental samples and the highest genetic variety was presented in those species with better dispersion capability or a smaller number of isolates.
SIGNIFICANCE AND IMPACT OF THE STUDY
This study permits to better understand the yeast communities in La Mancha region which gives a value the microbial potential of this region.
Topics: Biodiversity; Genetic Variation; Phylogeny; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Random Amplified Polymorphic DNA Technique; Rhodotorula; Saccharomycetales; Spain; Yeasts
PubMed: 32726883
DOI: 10.1111/jam.14795