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Journal of Fungi (Basel, Switzerland) Jun 2022Icariin is the most effective bioactive compound in Herba Epimedii. To enhance the content of icariin in the epimedium water extract, a novel strain, ZJU-L07, producing...
Icariin is the most effective bioactive compound in Herba Epimedii. To enhance the content of icariin in the epimedium water extract, a novel strain, ZJU-L07, producing an intracellular α-L-rhamnosidase was isolated from the soil and mutagenized. The specific activity of α-L-rhamnosidase was 29.89 U·mg through purification, and the molecular mass of the enzyme was 100 kDa, as assayed by SDS-PAGE. The characterization of the purified enzyme was determined. The optimal temperature and pH were 55 °C and 7.0, respectively. The enzyme was stable in the pH range 5.5-9.0 for 2 h over 80% and the temperature range 30-40 °C for 2 h more than 70%. The enzyme activity was inhibited by Ca, Fe, Cu, and Mg, especially Fe. The kinetic parameters of and were 1.38 mM and 24.64 μmol·mg·min using pNPR as the substrate, respectively. When epimedin C was used as a nature substrate to determine the kinetic parameters of α-L-rhamnosidase, the values of and were 3.28 mM and 0.01 μmol·mg·min, respectively. The conditions of enzymatic hydrolysis were optimized through single factor experiments and response surface methodology. The icariin yield increased from 61% to over 83% after optimization. The enzymatic hydrolysis method could be used for the industrialized production of icariin. At the same time, this enzyme could also cleave the α-1,2 glycosidic linkage between glucoside and rhamnoside in naringin and neohesperidin, which could be applicable in other biotechnological processes.
PubMed: 35736128
DOI: 10.3390/jof8060644 -
Applied and Environmental Microbiology Apr 2024Given the multitude of extracellular enzymes at their disposal, many of which are designed to degrade nature's polymers (lignin, cutin, cellulose, etc.), fungi are adept...
Given the multitude of extracellular enzymes at their disposal, many of which are designed to degrade nature's polymers (lignin, cutin, cellulose, etc.), fungi are adept at targeting synthetic polyesters with similar chemical composition. Microbial-influenced deterioration of xenobiotic polymeric surfaces is an area of interest for material scientists as these are important for the conservation of the underlying structural materials. Here, we describe the isolation and characterization of the 5307AH () cutinase, Plcut1. is basidiomycete yeast with the ability to disperse Impranil-DLN (Impranil), a colloidal polyester polyurethane, in agar plates. To test whether the fungal factor involved in this clearing was a secreted enzyme, we screened the ability of culture supernatants to disperse Impranil. Using size exclusion chromatography (SEC), we isolated fractions that contained Impranil-clearing activity. These fractions harbored a single ~22 kD band, which was excised and subjected to peptide sequencing. Homology searches using the peptide sequences identified, revealed that the protein Papla1 543643 (Plcut1) displays similarities to serine esterase and cutinase family of proteins. Biochemical assays using recombinant Plcut1 confirmed that this enzyme has the capability to hydrolyze Impranil, soluble esterase substrates, and apple cutin. Finally, we confirmed the presence of the Plcut1 in culture supernatants using a custom antibody that specifically recognizes this protein. The work shown here supports a major role for the Plcut1 in the fungal degradation of natural polyesters and xenobiotic polymer surfaces.IMPORTANCEFungi play a vital role in the execution of a broad range of biological processes that drive ecosystem function through production of a diverse arsenal of enzymes. However, the universal reactivity of these enzymes is a current problem for the built environment and the undesired degradation of polymeric materials in protective coatings. Here, we report the identification and characterization of a hydrolase from 5307AH, an aircraft-derived fungal isolate found colonizing a biodeteriorated polymer-coated surface. We show that secretes a cutinase capable of hydrolyzing soluble esters as well as ester-based compounds forming solid surface coatings. These findings indicate that this fungus plays a significant role in biodeterioration through the production of a cutinase adept at degrading ester-based polymers, some of which form the backbone of protective surface coatings. The work shown here provides insights into the mechanisms employed by fungi to degrade xenobiotic polymers.
PubMed: 38624219
DOI: 10.1128/aem.01694-23 -
Frontiers in Cellular and Infection... 2021Cryptococcosis is an infectious disease of worldwide distribution, caused by encapsulated yeasts belonging to the phylum Basidiomycota. The genus includes several...
Cryptococcosis is an infectious disease of worldwide distribution, caused by encapsulated yeasts belonging to the phylum Basidiomycota. The genus includes several species distributed around the world. The / species complex is largely responsible for most cases of cryptococcosis. However, clinical series have been published of infections caused by () and (), among other related genera. Here, we examined the pathogenic potential and antifungal susceptibility of / species complex (clades I and II) and related genera ( and ) isolated from environmental and clinical samples. . (clade III), . (clade IV) and /. (clade V) strains produced higher levels of phospholipase and hemolysins, whereas the / species complex strains (clades I and II) had markedly thicker capsules, produced more biofilm biomass and melanin, which are known virulence attributes. Interestingly, 40% of . strains (clade II) had MICs above the ECV established for this species to amphotericin B. Several non-/ species complex (clades III to V) had MICs equal to or above the ECVs established for and for all the three antifungal drugs tested. Finally, all the non- clinical isolates (clades III to V) produced more melanin than the environmental isolates might reflect their particularly enhanced need for melanin during protection. It is very clear that / species complex (clades I and II) strains, in general, show more similar virulence phenotypes between each other when compared to non-/ species complex (clades III to V) isolates. These observations together with the fact that . and spp. (clades III to V) strains were collected from the outside of a University Hospital, identify features of these yeasts important for environmental and patient colonization and furthermore, define mechanisms for infections with these uncommon pathogens.
Topics: Antifungal Agents; Basidiomycota; Cryptococcus gattii; Cryptococcus neoformans; Humans; Virulence; Virulence Factors
PubMed: 34277464
DOI: 10.3389/fcimb.2021.642658 -
PloS One 2022Neuromeningeal cryptococcosis (NMC) is a life-threatening opportunistic infection in advanced HIV disease patients (AHDP). It is caused by Cryptococcus spp. complexes...
Neuromeningeal cryptococcosis (NMC) is a life-threatening opportunistic infection in advanced HIV disease patients (AHDP). It is caused by Cryptococcus spp. complexes and mainly occurs in sub-Saharan Africa. In this study, we performed molecular characterization and antifungal susceptibility profiling of Cryptococcus isolates from AHDP in Kinshasa (DRC). Additionally, we investigated a possible association between NMC severity factors and the Cryptococcus neoformans (Cn) multilocus sequence typing (MLST) profiles. We characterized the isolates using PCR serotyping, MALDI-TOF MS, internal transcribed spacer (ITS) sequencing, and MLST. Susceptibility testing for the major antifungal drugs was performed according to the EUCAST guidelines. Parameters associated with NMC severity, such as hypoglycorrhachia (< 50 mg/dL), increased cerebral spinal fluid opening pressure (> 30 cm H2O), and poor therapeutic outcome were compared with the Cn MLST sequences type (ST). Twenty-three out of 29 Cryptococcus isolates were identified as serotype A using PCR serotyping (79.3%; 95% IC: 65.5-93.1), while six (20.7%; 95% IC: 6.9-34.5) were not serotypable. The 29 isolates were identified by ITS sequencing as follows: Cryptococcus neoformans (23/29, 79.3%), Cutaneotrichosporon curvatus (previously called Cryptococcus curvatus) (5/29, 17.2%), and Papiliotrema laurentii (Cryptococcus laurentii) (1/29, 3.5%). Using the ISHAM MLST scheme, all Cn isolates were identified as molecular type VNI. These comprised seven different STs: ST93 (n = 15), ST5 (n = 2), ST53 (n = 1), ST31 (n = 1), ST4 (n = 1), ST69 (n = 1), and one novel ST that has not yet been reported from other parts of the world and was subsequently assigned as ST659 (n = 2). Of the included strains, only Papiliotrema laurentii was resistant to amphoterin B (1/29, 3.5%), 6.8% (2/29) were resistant to 5-flucytosine (the single Papiliotrema laurentii strain and one Cryptococcus neoformans isolate), and 13.8% (4/29) to fluconazole, including two of five (40%) Cutaneotrichosporon curvatus and two of 23 (8.7%) C. neoformans strains. We found a significative association between poor therapeutic outcome and a non-ST93 sequence type of causative strains (these concerned the less common sequence types: ST53, ST31, ST5, ST4, ST659, and ST69) (87.5% versus 40%, p = 0.02). Molecular analysis of Cryptococcus spp. isolates showed a wide species diversity and genetic heterogenicity of Cn within the VNI molecular type. Furthermore, it is worrying that among included strains we found resistances to several of the commonly used antifungals.
Topics: Antifungal Agents; Basidiomycota; Cryptococcosis; Cryptococcus neoformans; Democratic Republic of the Congo; Genetic Variation; Genotype; HIV Infections; Humans; Multilocus Sequence Typing; Mycological Typing Techniques
PubMed: 35587939
DOI: 10.1371/journal.pone.0267842 -
G3 (Bethesda, Md.) Dec 2021Papiliotrema terrestris strain LS28 is a biocontrol agent selected for its antagonistic activity against several plant pathogens both in the field and postharvest. The...
Papiliotrema terrestris strain LS28 is a biocontrol agent selected for its antagonistic activity against several plant pathogens both in the field and postharvest. The availability of a genome sequencing sets the foundation for the identification of the genetic mechanisms of its antagonistic activity. The genome size is 21.29 Mbp with a G+C content of 58.65%, and genome annotation predicts 8,626 protein-encoding genes. Phylogenetic analysis based on whole-genome data confirms that P. terrestris is a Tremellomycetes more closely related to Papiliotrema flavescens than Papiliotrema laurentii.
Topics: Basidiomycota; High-Throughput Nucleotide Sequencing; Phylogeny; Saccharomyces cerevisiae; Sequence Analysis, DNA
PubMed: 34534326
DOI: 10.1093/g3journal/jkab332 -
The Brazilian Journal of Infectious... 2019Papiliotrema laurentii is one of several non-neoformans cryptococci that have rarely been associated with human infection, since it was previously considered saprophyte...
BACKGROUND
Papiliotrema laurentii is one of several non-neoformans cryptococci that have rarely been associated with human infection, since it was previously considered saprophyte and thought to be non-pathogenic to humans. Nevertheless, increasing number of reports of human infection have emerged in recent years, mostly in oncologic patients.
AIM
To report a case of a female patient with pyloric obstructive cancer with a catheter-related Papiliotrema laurentii blood stream infection and systematically review the available evidence on P. laurentii infection in humans.
METHODS
Retrieval of studies was based on Medical Subject Headings and Health Sciences Descriptors, which were combined using Boolean operators. Searches were run on the electronic databases Scopus, Web of Science, MEDLINE (PubMed), BIREME (Biblioteca Regional de Medicina), LILACS (Latin American and Caribbean Health Sciences Literature), Cochrane Library for Systematic Reviews and Opengray.eu. There was no language or date of publication restrictions. The reference lists of the studies retrieved were searched manually.
RESULTS
The search strategy retrieved 1703 references. In the final analysis, 31 references were included, with the description of 35 cases. Every patient but one had a previous co-morbidity - 48.4 % of patients had a neoplasm. Amphotericin B was the most used treatment and only a single case of resistance to it was reported. Most patients were cured of the infection.
CONCLUSION
P. laurentii infection in humans is usually associated to neoplasia and multiple co-morbidities, and amphotericin B seems to be a reliable agent for treatment.
Topics: Aged; Amphotericin B; Anti-Bacterial Agents; Bacteremia; Biopsy; Catheter-Related Infections; Cryptococcus; Female; Fluconazole; Humans; Piperacillin, Tazobactam Drug Combination; Stomach Neoplasms; Tomography, X-Ray Computed; Vancomycin
PubMed: 31738886
DOI: 10.1016/j.bjid.2019.10.005 -
3 Biotech Sep 2020In this work, we isolated and selected oleaginous yeasts from rock field soils from two National Parks in Brazil ( and ) with the potential to accumulate oil from...
In this work, we isolated and selected oleaginous yeasts from rock field soils from two National Parks in Brazil ( and ) with the potential to accumulate oil from xylose, the main pentose sugar found in lignocellulosic biomass. From the 126 isolates, two were selected based on their lipid contents. They were taxonomically identified as (UFV-1 and UFV-2). Of the two, UFV-1 was selected as the best lipid producer Under unoptimized conditions, lipid production by UFV-1 was higher in glucose than in xylose. To improve its lipid production from xylose, we applied response surface methodology (RSM) with a face-centered central composite design (CCF). We evaluated the effects of agitation rate, initial cell biomass (OD), carbon/nitrogen ratio (/ ratio) and pH on lipid production. UFV-1 recorded the highest lipid content, 63.5% (w/w) of the cell dry mass, under the following conditions: / ratio = 100:1, pH value = 7.0, initial OD = 0.8 and agitation = 300 rpm. Under these optimized conditions, biomass, lipid titer and volumetric lipid productivity were 9.31 g/L, 5.90 g/L and 0.082 g/L.h, respectively. Additionally, we determined the fatty acid composition of UFV-1 as follows: C14:0 (0.5%), C16:0 (28.4-29.4%), C16:1 (0.2%), C18:0 (9.5-11%), C18:1 (58.6-60.5%), and C20:0 (0.7-0.8%). Based on this composition, the predicted properties of biodiesel showed that UFV-1 oil is suitable for use as feedstock in biodiesel production.
PubMed: 32802724
DOI: 10.1007/s13205-020-02373-4 -
Fungal Genetics and Biology : FG & B Jan 2023The production of yeast oil from lignocellulosic biomasses is impaired by inhibitors formed during the pretreatment step, mainly acetic acid. Herein, we applied Adaptive...
The production of yeast oil from lignocellulosic biomasses is impaired by inhibitors formed during the pretreatment step, mainly acetic acid. Herein, we applied Adaptive Laboratory Evolution (ALE) to select three Acetic acid Tolerant Strains (ATS) of P. laurentii UFV-1. Different phenotypes emerged alongside evolution. The ATS II presented trade-offs in the absence of acetic acid, suggesting that it displays a specialized phenotype of tolerance to growth on organic acids. On the other hand, ATS I and ATS III presented phenotypes associated with the behavior of generalists. ATS I was considered the most promising evolved strain as it displayed the oleaginous phenotype in all conditions tested. Thus, we applied whole-genome sequencing to detect the mutations that emerged in this strain during the ALE. We found alterations in genes encoding proteins involved in different cellular functions, including multidrug resistance (MDR) transporters, energy metabolism, detoxification, coenzyme recycling, and cell envelope remodeling. To evaluate acetic acid stress responses, both parental and ATS I strains were cultivated in chemostat mode in the absence and presence of acetic acid. In contrast to ATS I, the parental strain presented alterations in the cell envelope and cell size under acetic acid stress conditions. Furthermore, the parental strain and the ATS I presented differences regarding acetic acid assimilation. Contrary to the parental strain, the ATS I displayed an increase in unsaturated fatty acid content irrespective of acetic acid stress, which might be related to improved tolerance to acetic acid. Altogether, these results provided insights into the mechanisms involved with the acetic acid tolerance displayed by ATS I and the responses of P. laurentii to this stressful condition.
Topics: Acetic Acid; Saccharomyces cerevisiae; Phenotype; Membrane Transport Proteins
PubMed: 36528339
DOI: 10.1016/j.fgb.2022.103765 -
BMJ Case Reports May 2022A man in mid-50s presented with progressive blurred vision in his left eye for over 6 weeks. He was a known diabetic with history of COVID-19 pneumonia treated with...
A man in mid-50s presented with progressive blurred vision in his left eye for over 6 weeks. He was a known diabetic with history of COVID-19 pneumonia treated with steroids and remdesivir. He had pyelonephritis and urinary culture grown He was referred as a case of non-resolving vitreous haemorrhage. Visual acuity (VA) was hand movements with fundus showing dense vitritis. He underwent pars plana vitrectomy, vitreous biopsy with intraocular antibiotics (imipenem) suspecting as a case of endogenous bacterial endophthalmitis. Vitreous biopsy did not yield organisms on the smear/culture. The patient's condition worsened with perception of light and fundus showing dense vitritis with discrete yellowish white deposits on the surface of the retina. A repeat vitreous biopsy done along with intravitreal injection of voriconazole (suspecting fungal aetiology) grown fungal colonies and the organism was identified as At 4-month follow-up, the VA improved to 6/24.
Topics: Anti-Bacterial Agents; Basidiomycota; COVID-19; Endophthalmitis; Eye Infections, Bacterial; Humans; Male; Retrospective Studies; Vitrectomy
PubMed: 35545307
DOI: 10.1136/bcr-2021-246637 -
Scientific Reports Sep 2023The Cryptococcus genus comprises more than 100 species, of which C. neoformans and C. gattii are the leading cause of cryptococcosis. The distribution of C. gattii and...
The Cryptococcus genus comprises more than 100 species, of which C. neoformans and C. gattii are the leading cause of cryptococcosis. The distribution of C. gattii and C. neoformans species complexes has been extensively studied and widely reported globally. Other species such as Naganishia albida, Papiliotrema laurentii, and Papiliotrema flavescens have been reported as pathogenic yeasts. Since there are no reports of environmental isolation in the Boyacá region (Colombia), this study aimed to isolate and characterize Cryptococcus and Cryptococcus-like yeasts from pigeon feces, Eucalyptus, and olive trees distributed in the municipalities of Tunja and Ricaute Alto. The environmental data was recovered, and the isolations obtained were identified by microscopy, biochemical test, MALDI-TOF MS, URA5-RFLP, and sequencing of the ITS and LSU loci. For the 93 pigeon dropping samples collected in Tunja, 23 yielded to C. neoformans, 3 to N. globosa, 2 N. albida and 1 to P. laurentii. Of the 1188 samples collected from olive trees, 17 (1.43%) positive samples were identified as C. gattii species complex (4), C. neoformans species complex (2), P. laurentii (3), N. albida (2), N. globosa (5) and P. flavescens (1). Likewise, specimens of C. neoformans presented molecular type VNI and molecular type VNII; for C. gattii the molecular types found were VGIII and one VGIV by URA5-RFLP but VGIII by MALDI-TOF and sequencing of the ITS and LSU. Therefore, it can be concluded that the species of Cryptococcus, Naganishia and Papiliotrema genera, are present in the environment of Boyacá, and show a predilection for climate conditions that are typical of this region.
Topics: Animals; Colombia; Cryptococcus neoformans; Cryptococcus gattii; Cryptococcosis; Climate; Columbidae; Olea
PubMed: 37735454
DOI: 10.1038/s41598-023-41994-6