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Journal of Fungi (Basel, Switzerland) Mar 2021was analyzed for the production of secondary metabolites, resulting in the isolation of known zopfinol () and its new derivatives zopfinol B-C (), the 10-membered...
was analyzed for the production of secondary metabolites, resulting in the isolation of known zopfinol () and its new derivatives zopfinol B-C (), the 10-membered lactones 7-O-acetylmultiplolide A () and 8-O-acetylmultiplolide A (), together with sordarin (), sordarin B (), and hypoxysordarin (). The absolute configuration of was elucidated by the synthesis of MPTA-esters. Compound showed antimicrobial activity against the Gram-positive bacteria and and the fungus . While was weakly antibacterial, showed stronger antibiotic activity against the Gram-positive bacteria and weak antifungal activity against and . We furthermore observed the cytotoxicity of , and against the mammalian cell lines KB3.1 and L929. Moreover, the new genus is introduced herein to accommodate together with several species of -, , and . These taxa formed a well-supported monophyletic clade in the recently introduced family Navicularisporaceae, located far from the type species of the respective original genera, in a phylogram based on the combined dataset sequences of the internal transcribed spacer region (ITS), the nuclear rDNA large subunit (LSU), and fragments of the ribosomal polymerase II subunit 2 () and β-tubulin () genes. is synonymized with due to the phylogenetic and morphological similarity. The isolation of zopfinols - and sordarins - confirms the potential of this fungal order as producers of bioactive compounds and suggests these compounds as potential chemotaxonomic markers.
PubMed: 33802411
DOI: 10.3390/jof7030181 -
Drug Delivery Dec 2019The potential advantages of recombinant microbes as oral drug carriers for curing diseases have attracted much attention. The use of recombinant oil microbes as living...
The potential advantages of recombinant microbes as oral drug carriers for curing diseases have attracted much attention. The use of recombinant oil microbes as living cell liposomes to carry polypeptide drugs may be an ideal polypeptide oral drug delivery system. GM4-ΔTS was constructed by LFH-PCR from Rhodotorula glutinis GM4, which was screened and preserved in our laboratory, and then transferred into choline-phosphate cytidylyltransferase (CCT), which is a rate-limiting enzyme for lecithin synthesis. The results showed that the CCT gene was highly expressed in the GM4-ΔTS strain and could significantly increase fatty acid and lecithin contents in GM4-ΔTS-PGK1-CCT. Moreover, insulin, H22-LP, and α-MSH were successfully introduced into cells in vitro, and the strain no longer proliferated in vivo, for safe and controllable polypeptide drug delivery. In vivo, normal mice were intragastrically administered with recombinant strains carrying insulin and α-MSH, and different levels of polypeptide drugs were detected in serum and tissue, respectively. Then, recombinant strains carrying insulin were administered to type II diabetes mellitus mice. The results showed that the strains could effectively reduce blood glucose levels in mice, which indicated that the recombinant strains could carry insulin into the body, and the drug effect was remarkable. Therefore, recombinant GM4-ΔTS-PGK1-CCT strains were successfully used as living cell liposomes to carry insulin, H22-LP, and α-MSH peptides into the body for the first time; additionally, these strains have enhanced safety, controllability, and efficacy.
Topics: Animals; Diabetes Mellitus, Experimental; Drug Carriers; Insulin; Liposomes; Mice; Mice, Inbred BALB C; Peptides; Random Allocation; Rhodotorula
PubMed: 30744426
DOI: 10.1080/10717544.2018.1551439 -
BMC Genomics Nov 2020Rhodotorula glutinis is recognized as a biotechnologically important oleaginous red yeast, which synthesizes numerous meritorious compounds with wide industrial usages....
Genomics and lipidomics analysis of the biotechnologically important oleaginous red yeast Rhodotorula glutinis ZHK provides new insights into its lipid and carotenoid metabolism.
BACKGROUND
Rhodotorula glutinis is recognized as a biotechnologically important oleaginous red yeast, which synthesizes numerous meritorious compounds with wide industrial usages. One of the most notable properties of R. glutinis is the formation of intracellular lipid droplets full of carotenoids. However, the basic genomic features that underlie the biosynthesis of these valuable compounds in R. glutinis have not been fully documented. To reveal the biotechnological potential of R. glutinis, the genomics and lipidomics analysis was performed through the Next-Generation Sequencing and HPLC-MS-based metabolomics technologies.
RESULTS
Here, we firstly assemble the genome of R. glutinis ZHK into 21.8 Mb, containing 30 scaffolds and 6774 predicted genes with a N50 length of 14, 66,672 bp and GC content of 67.8%. Genome completeness assessment (BUSCO alignment: 95.3%) indicated the genome assembly with a high-quality features. According to the functional annotation of the genome, we predicted several key genes involved in lipids and carotenoids metabolism as well as certain industrial enzymes biosynthesis. Comparative genomics results suggested that most of orthologous genes have underwent the strong purifying selection within the five Rhodotorula species, especially genes responsible for carotenoids biosynthesis. Furthermore, a total of 982 lipids were identified using the lipidomics approaches, mainly including triacylglycerols, diacylglyceryltrimethylhomo-ser and phosphatidylethanolamine.
CONCLUSION
Using whole genome shotgun sequencing, we comprehensively analyzed the genome of R. glutinis and predicted several key genes involved in lipids and carotenoids metabolism. By performing comparative genomic analysis, we show that most of the ortholog genes have undergone strong purifying selection within the five Rhodotorula species. Furthermore, we identified 982 lipid species using lipidomic approaches. These results provided valuable resources to further advance biotechnological applications of R .glutinis.
Topics: Biological Products; Carotenoids; Genomics; Lipidomics; Lipids; Rhodotorula
PubMed: 33243144
DOI: 10.1186/s12864-020-07244-z -
Bioengineered Dec 2019Selenium-enriched yeast can transform toxic inorganic selenium into absorbable organic selenium, which is of great significance for human health and pharmaceutical...
Selenium-enriched yeast can transform toxic inorganic selenium into absorbable organic selenium, which is of great significance for human health and pharmaceutical industry. A yeast X-20 we obtained before has good selenium-enriched ability, but its selenium content is still low for industrial application. In this study, strategies of process optimization and transport regulation of selenium were thus employed to further improve the cell growth and selenium enrichment. Through engineering phosphate transporters from into X-20, the selenium content was increased by 21.1%. Through using mixed carbon culture (20 g L, glycerol: glucose 3:7), both biomass and selenium content were finally increased to 5.3 g L and 5349.6 µg g (cell dry weight, DWC), which were 1.14 folds and 6.77 folds compared to their original values, respectively. Our results indicate that high selenium-enrichment ability and biomass production can be achieved through combining process optimization and regulation of selenium transport.
Topics: Biological Transport; Biomass; Culture Media; Fermentation; Gene Expression; Glucose; Glycerol; Metabolic Engineering; Phosphate Transport Proteins; Phosphates; Plasmids; Proton-Phosphate Symporters; Rhodotorula; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins; Selenium; Sodium-Phosphate Cotransporter Proteins, Type III; Transgenes
PubMed: 31322471
DOI: 10.1080/21655979.2019.1644853 -
Biotechnology For Biofuels 2019Lipid extraction for quantification of fat content in oleaginous yeasts often requires strong acids and harmful organic solvents; it is laborious and time-consuming....
BACKGROUND
Lipid extraction for quantification of fat content in oleaginous yeasts often requires strong acids and harmful organic solvents; it is laborious and time-consuming. Therefore, in most cases just endpoint measurements of lipid accumulation are performed and kinetics of intracellular lipid accumulation is difficult to follow. To address this, we created a prediction model using Fourier-transform near-infrared (FT-NIR) spectroscopy. This method allows to measure lipid content in yeast.
METHODS
The FT-NIR calibration sets were constructed from spectra of freeze-dried cells of the oleaginous yeasts CBS 14, CBS 1807 and CBS 6114. The yeast cells were obtained from different cultivation conditions. Freeze-dried cell pellets were scanned using FT-NIR in the Multi Purpose Analyser (MPA) from Bruker. The obtained spectra were assigned corresponding to total fat content, obtained from lipid extraction using a modified Folch method. Quantification models using partial least squares (PLS) regression were built, and the calibration sets were validated on independently cultivated samples. The model was additionally tested on DBVPG 8058 and CBS 2387.
RESULTS
The of the FT-NIR model for was 98%, and the root mean square error of cross-validation (RMSECV) was 1.53. The model was validated using a separate set of samples with a root mean square error of prediction (RMSEP) of 3.21. The of the model was 96%, with RMSECV 2.4 and RMSEP 3.8. The of the mixed model, including all tested yeast strains, was 90.5%, with RMSECV 2.76 and RMSEP 3.22, respectively. The models were verified by predicting the total fat content in newly cultivated and freeze-dried samples. Additionally, the kinetics of lipid accumulation of a culture were followed and compared with standard lipid extraction methods.
CONCLUSIONS
Using FT-NIR spectroscopy, we have developed a faster, less laborious and non-destructive quantification of yeast intracellular lipid content compared to methods using lipid extraction.
PubMed: 31297157
DOI: 10.1186/s13068-019-1513-9 -
Antonie Van Leeuwenhoek Nov 2022Analysis of predicted fungal proteomes revealed a large family of sequences that showed similarity to the Saccharomyces cerevisiae Class-I dihydroorotate dehydrogenase...
Analysis of predicted fungal proteomes revealed a large family of sequences that showed similarity to the Saccharomyces cerevisiae Class-I dihydroorotate dehydrogenase Ura1, which supports synthesis of pyrimidines under aerobic and anaerobic conditions. However, expression of codon-optimised representatives of this gene family, from the ascomycete Alternaria alternata and the basidiomycete Schizophyllum commune, only supported growth of an S. cerevisiae ura1Δ mutant when synthetic media were supplemented with dihydrouracil. A hypothesis that these genes encode NAD(P)-dependent dihydrouracil dehydrogenases (EC 1.3.1.1 or 1.3.1.2) was rejected based on absence of complementation in anaerobic cultures. Uracil- and thymine-dependent oxygen consumption and hydrogen-peroxide production by cell extracts of S. cerevisiae strains expressing the A. alternata and S. commune genes showed that, instead, they encode active dihydrouracil oxidases (DHO, EC1.3.3.7). DHO catalyses the reaction dihydrouracil + O → uracil + HO and was only reported in the yeast Rhodotorula glutinis (Owaki in J Ferment Technol 64:205-210, 1986). No structural gene for DHO was previously identified. DHO-expressing strains were highly sensitive to 5-fluorodihydrouracil (5F-dhu) and plasmids bearing expression cassettes for DHO were readily lost during growth on 5F-dhu-containing media. These results show the potential applicability of fungal DHO genes as counter-selectable marker genes for genetic modification of S. cerevisiae and other organisms that lack a native DHO. Further research should explore the physiological significance of this enigmatic and apparently widespread fungal enzyme.
Topics: Saccharomyces cerevisiae; Hydrogen Peroxide; Thymine; Proteome; Cell Extracts; NAD; Genes, Fungal; Uracil; Hydrogen
PubMed: 36241945
DOI: 10.1007/s10482-022-01779-9 -
Journal of Fungi (Basel, Switzerland) Apr 2023During a study of the diversity of soilborne fungi from Spain, a strain belonging to the family Chaetomiaceae (Sordariales) was isolated. The multigene phylogenetic...
During a study of the diversity of soilborne fungi from Spain, a strain belonging to the family Chaetomiaceae (Sordariales) was isolated. The multigene phylogenetic inference using five DNA loci showed that this strain represents an undescribed species of the genus , herein introduced as sp. nov. Investigation of its secondary metabolome led to the isolation of two new derivatives ( and ) of the known antifungal antibiotic dactylfungin A (), together with the known compound cochliodinol (). The planar structures of - were determined by ultrahigh performance liquid chromatography coupled with diode array detection and ion mobility tandem mass spectrometry (UHPLC-DAD-IM-MS/MS) and extensive 1D and 2D nuclear magnetic resonance (NMR) spectroscopy after isolation by HPLC. All isolated secondary metabolites were tested for their antimicrobial and cytotoxic activities. Dactylfungin A () showed selective and strong antifungal activity against some of the tested human pathogens ( and ). The additional hydroxyl group in resulted in the loss of activity against but still retained the inhibition of in a lower concentration than that of the respective control, without showing any cytotoxic effects. In contrast, 25″-dehydroxy-dactylfungin A () exhibited improved activity against yeasts ( and ) than and , but resulted in the appearance of slight cytotoxicity. The present study exemplifies how even in a well-studied taxonomic group such as the Chaetomiaceae, the investigation of novel taxa still brings chemistry novelty, as demonstrated in this first report of this antibiotic class for chaetomiaceous and sordarialean taxa.
PubMed: 37108917
DOI: 10.3390/jof9040463 -
Frontiers in Chemistry 2024This study investigates the biological activities of essential oil (LPEO), an endemic lavender species from the Canary Islands, traditionally used in treating various...
This study investigates the biological activities of essential oil (LPEO), an endemic lavender species from the Canary Islands, traditionally used in treating various ailments. LPEO was extracted by hydrodistillation and analyzed using GC-MS. Antioxidant activity was assessed by DPPH radical scavenging and total antioxidant capacity assays. Antimicrobial activity was evaluated by disc diffusion, MIC, MBC, and MFC determination against bacterial () and fungal () strains. Antidiabetic and anti-gout potential were investigated through α-amylase, α-glucosidase, and xanthine oxidase inhibition assays. Antityrosinase activity was determined using a modified dopachrome method. Cytotoxicity was assessed by MTT assay against breast (MCF-7, MDA-MB-468), liver (HepG2), colon (HCT-15) cancer cells, and normal cells (PBMCs). LPEO exhibits potent antiradical activity (IC50 = 148.33 ± 2.48 μg/mL) and significant antioxidant capacity (TAC = 171.56 ± 2.34 μg AA/mg of EO). It demonstrates notable antibacterial activity against four strains ( and ) with inhibition zones ranging from 18.70 ± 0.30 mm to 29.20 ± 0.30 mm, along with relatively low MIC and MBC values. LPEO displays significant antifungal activity against four strains ( and ) with a fungicidal effect at 1 mg/mL, surpassing the positive control (cycloheximide), and MIC and MFC values indicating a fungicidal effect. It exhibits substantial inhibition of xanthine oxidase enzyme (IC50 = 26.48 ± 0.90 μg/mL), comparable to allopurinol, and marked inhibitory effects on α-amylase (IC50 = 31.56 ± 0.46 μg/mL) and α-glucosidase (IC50 = 58.47 ± 2.35 μg/mL) enzymes.The enzyme tyrosinase is inhibited by LPEO (IC50 = 29.11 ± 0.08 mg/mL). LPEO displays moderate cytotoxic activity against breast, liver, and colon cancer cells, with low toxicity towards normal cells (PBMC). LPEO exhibits greater selectivity than cisplatin for breast (MCF-7) and colon (HCT-15) cancer cells but lower selectivity for liver (HepG2) and metastatic breast (MDA-MB-468) cancer cells. These findings suggest the potential of LPEO as an antioxidant, antimicrobial, anti-gout, antidiabetic, and anticancer agent.
PubMed: 38660570
DOI: 10.3389/fchem.2024.1383731 -
Open Veterinary Journal Jun 2023The fungi species are widespread airborne contaminants and are thought to be natural occupants of human skin, lungs, urine, and feces. Therefore, , , and are three...
BACKGROUND
The fungi species are widespread airborne contaminants and are thought to be natural occupants of human skin, lungs, urine, and feces. Therefore, , , and are three of the most prevalent species.
AIM
This study aims to isolate from the rumen fluid of cows in the province of Mosul and to determine how laser light irradiation affects the growth and morphological traits of these Fungi.
METHODS
From the rumen fluid of AL-Restaki and AL-Karadi of cows, the was isolated. Using the traditional approach and the ID-Yst card system Vitek 2. A semiconductor laser system with a power of 50 mW and a wavelength of 450 nm was used in the experiment to evaluate the light laser irradiation effects on the culture growth of directly under two light irradiation conditions of 30 and 60 minutes.
RESULTS
According to traditional methods and the ID-Yst card system Vitek 2, predominated 7/30 (23.3%), and these strains effectively grow on medium sabouraued dextrose agar as evidenced by the carotenoid pigments that gave their colonies a salmon-pink to coral-red. Compared with a control group where no laser was used, the impact of light laser irradiation was assessed 24 hours after the irradiation using biomass (dry weight measuring yeast cell content in suspension) and microscopic analysis using Gram stain. Microscopic examinations showed the irregular shape of the cells linked to one another. The irradiated subculture of on Sabouraued dextrose agar and incubation at 37°C for 3 days demonstrated inhibited growth in 4/7 (57.1%) isolates. In addition, there was no discernible difference vertically at < 0.05 between the control group and the biomass concentration under light irradiation circumstances (30 and 60 minutes).
CONCLUSION
This study proved that is found in the rumen fluid of cows. Also, the isolated displayed sensitivity to laser irradiation lights, revealing the more significant topographical alterations of the cell structure that had happened, the irregular shape of the cells, and how they were connected as a result of evolution.
Topics: Cattle; Animals; Humans; Rhodotorula; Agar; Iraq; Glucose
PubMed: 37545703
DOI: 10.5455/OVJ.2023.v13.i6.11 -
Foods (Basel, Switzerland) Aug 2021In the last decades, several plant-based materials were used for the substitution of fish meal and oil in aquaculture. The present study evaluated the fish quality and...
In the last decades, several plant-based materials were used for the substitution of fish meal and oil in aquaculture. The present study evaluated the fish quality and the sensory differences of rainbow trout () and pike-perch () from three different feeding groups, which were fed a commercially available industrial (standard) diet, a control diet, and a special microorganism-based feed mix. This feed mainly consisted of a mix made of and sp. and had 50% less fish meal and fish oil compared to typical control diets. At the beginning, the pike-perch population was six months old, and the rainbow trout population was 15 months old. The feeding study duration was 16 weeks and every four weeks the growth performance and several morphometric parameters were recorded. Afterwards, sensory evaluation took place to identify possible trends. Sensory evaluation revealed that the rainbow trout groups did not show any significant differences to the standard and control fish fillets with regard to odor, texture, and taste. The effects on rainbow trout growth performances and carcass parameters were similar to the standard group. The feed mix was not optimal for pike-perch farming, which was also reflected by significantly adversely affected growth performance and carcass parameters. The sensorial evaluation showed an opposite trend: here, only small differences in the fillets from the feed mix and standard/control diet were observed.
PubMed: 34441576
DOI: 10.3390/foods10081799