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Maedica Mar 2022is a widely distributed fungus that has evolved as an important pathogen, especially in immunocompromised individuals, causing fungemias, central nervous system...
is a widely distributed fungus that has evolved as an important pathogen, especially in immunocompromised individuals, causing fungemias, central nervous system infections, peritoneal dialysis-associated peritonitis and keratitis. Even though there are a few reports of skin and soft tissue infections caused by , all these cases involve immunocompromised individuals. is a Gram-positive bacterium known to cause several infections such as bloodstream, urinary tract and skin and soft tissue infections. We report a mixed and skin and soft tissue infection in a 63-year-old woman with well controlled type 2 diabetes mellitus and no other known history of immunosuppression, suffering from skin and soft tissue infection of the right lower extremity. The patient did not respond to treatment with broad spectrum antimicrobials, but had a successful outcome with fluconazole, after was isolated from pus of the skin ulcer. The antifungal treatment led to eradication of the infection, while no recurrence was observed during a follow-up period of two years. Clinicians should be aware that can cause infection even in immunocompetent patients.
PubMed: 35733763
DOI: 10.26574/maedica.2022.17.1.211 -
Applied Microbiology and Biotechnology Jul 2016Rhodotorula glutinis is capable of synthesizing numerous valuable compounds with a wide industrial usage. Biomass of this yeast constitutes sources of microbiological... (Review)
Review
Rhodotorula glutinis is capable of synthesizing numerous valuable compounds with a wide industrial usage. Biomass of this yeast constitutes sources of microbiological oils, and the whole pool of fatty acids is dominated by oleic, linoleic, and palmitic acid. Due to its composition, the lipids may be useful as a source for the production of the so-called third-generation biodiesel. These yeasts are also capable of synthesizing carotenoids such as β-carotene, torulene, and torularhodin. Due to their health-promoting characteristics, carotenoids are commonly used in the cosmetic, pharmaceutical, and food industries. They are also used as additives in fodders for livestock, fish, and crustaceans. A significant characteristic of R. glutinis is its capability to produce numerous enzymes, in particular, phenylalanine ammonia lyase (PAL). This enzyme is used in the food industry in the production of L-phenylalanine that constitutes the substrate for the synthesis of aspartame-a sweetener commonly used in the food industry.
Topics: Biofuels; Biomass; Carotenoids; Enzymes; Fatty Acids; Industrial Microbiology; Linoleic Acid; Oleic Acid; Palmitic Acid; Phenylalanine; Phenylalanine Ammonia-Lyase; Rhodotorula; beta Carotene
PubMed: 27209039
DOI: 10.1007/s00253-016-7611-8 -
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 -
Frontiers in Nutrition 2022, as a member of the family , is of great value in the field of biotechnology. However, the evolutionary relationship of X-20 with , , and are not well understood, and...
, as a member of the family , is of great value in the field of biotechnology. However, the evolutionary relationship of X-20 with , , and are not well understood, and its metabolic pathways such as carotenoid biosynthesis are not well resolved. Here, genome sequencing and comparative genome techniques were employed to improve the understanding of X-20. Phytoene desaturase (crtI) and 15-cis-phytoene synthase/lycopene beta-cyclase (crtYB), key enzymes in carotenoid pathway from X-20 were more efficiently expressed in INVSc1 than in CEN.PK2-1C. High yielding engineered strains were obtained by using synthetic biology technology constructing carotenoid pathway in and optimizing the precursor supply after fed-batch fermentation with palmitic acid supplementation. Genome sequencing analysis and metabolite identification has enhanced the understanding of evolutionary relationships and metabolic pathways in X-20, while heterologous construction of carotenoid pathway has facilitated its industrial application.
PubMed: 35782944
DOI: 10.3389/fnut.2022.918240 -
Microbial Cell Factories Aug 2022The regioselective hydroxylation of phenolic compounds, especially flavonoids, is still a bottleneck of classical organic chemistry that could be solved using enzymes...
BACKGROUND
The regioselective hydroxylation of phenolic compounds, especially flavonoids, is still a bottleneck of classical organic chemistry that could be solved using enzymes with high activity and specificity. Yeast Rhodotorula glutinis KCh735 in known to catalyze the C-8 hydroxylation of flavones and flavanones. The enzyme F8H (flavonoid C8-hydroxylase) is involved in the reaction, but the specific gene has not yet been identified. In this work, we present identification, heterologous expression and characterization of the first F8H ortho-hydroxylase from yeast.
RESULTS
Differential transcriptome analysis and homology to bacterial monooxygenases, including also a FAD-dependent motif and a GD motif characteristic for flavin-dependent monooxygenases, provided a set of coding sequences among which RgF8H was identified. Phylogenetic analysis suggests that RgF8H is a member of the flavin monooxygenase group active on flavonoid substrates. Analysis of recombinant protein showed that the enzyme catalyzes the C8-hydroxylation of naringenin, hesperetin, eriodyctiol, pinocembrin, apigenin, luteolin, chrysin, diosmetin and 7,4'-dihydroxyflavone. The presence of the C7-OH group is necessary for enzymatic activity indicating ortho-hydroxylation mechanism. The enzyme requires the NADPH coenzyme for regeneration prosthetic group, displays very low hydroxyperoxyflavin decupling rate, and addition of FAD significantly increases its activity.
CONCLUSIONS
This study presents identification of the first yeast hydroxylase responsible for regioselective C8-hydroxylation of flavonoids (F8H). The enzyme was biochemically characterized and applied in in vitro cascade with Bacillus megaterium glucose dehydrogenase reactions. High in vivo activity in Escherichia coli enable further synthetic biology application towards production of rare highly antioxidant compounds.
Topics: Flavin-Adenine Dinucleotide; Flavins; Flavonoids; Hydroxylation; Mixed Function Oxygenases; Phylogeny; Rhodotorula; Substrate Specificity
PubMed: 36038906
DOI: 10.1186/s12934-022-01899-x -
Scientific Reports Jul 2018Rhodotorula glutinis, an oleaginous red yeast, intrinsically produces several bio-products (i.e., lipids, carotenoids and enzymes) and is regarded as a potential host...
Rhodotorula glutinis, an oleaginous red yeast, intrinsically produces several bio-products (i.e., lipids, carotenoids and enzymes) and is regarded as a potential host for biorefinery. In view of the limited available genetic engineering tools for this yeast, we have developed a useful genetic transformation method and transformed the β-carotene biosynthesis genes (crtI, crtE, crtYB and tHMG1) and cellulase genes (CBHI, CBHII, EgI, EgIII, EglA and BGS) into R. glutinis genome. The transformant P4-10-9-63Y-14B produced significantly higher β-carotene (27.13 ± 0.66 mg/g) than the wild type and also exhibited cellulase activity. Furthermore, the lipid production and salt tolerance ability of the transformants were unaffected. This is the first study to engineer the R. glutinis for simultaneous β-carotene and cellulase production. As R. glutinis can grow in sea water and can be engineered to utilize the cheaper substrates (i.e. biomass) for the production of biofuels or valuable compounds, it is a promising host for biorefinery.
Topics: Biofuels; Cellulase; Fungal Proteins; Gene Expression Regulation, Fungal; Genetic Engineering; Genome, Fungal; Industrial Microbiology; Rhodotorula; beta Carotene
PubMed: 30022171
DOI: 10.1038/s41598-018-29194-z -
Ultrasonics Sonochemistry Mar 2020The extraction of Rhodotorula glutinis carotenoids by ultrasound under pressure (manosonication) in an aqueous medium has been demonstrated. The influence of treatment...
The extraction of Rhodotorula glutinis carotenoids by ultrasound under pressure (manosonication) in an aqueous medium has been demonstrated. The influence of treatment time, pressure, and ultrasound amplitude on R. glutinis inactivation and on the extraction of carotenoids was evaluated, and the obtained data were described mathematically. The extraction yields were lineal functions of those three parameters, whereas inactivation responded to a more complex equation. Under optimum treatment conditions, 82% of carotenoid content was recovered. Extraction of carotenoids in an aqueous medium was attributed to the capacity of ultrasound for cell disruption and emulsification. Cavitation caused the rupture of cell envelopes and the subsequent formation of small droplets of carotenoids surrounded by the phospholipids of the cytoplasmic membrane that would stabilize the emulsion. Analysis of the dispersed particle size of the extracts demonstrated that a fine, homogeneous emulsion was formed after treatment (average size: 230 nm; polydispersity <0.22). This research describes an innovative green process for extracting carotenoids from fresh biomass of R. glutinis in which only two unit operations are required: ultrasonic treatment, followed by a centrifugation step to discard cell debris. The extract obtained thanks to this procedure is rich in carotenoids (25 mg/L) and could be directly incorporated as a pigment in foods, beverages, and diet supplements; it can also be utilized as an ingredient in drugs or cosmetics.
Topics: Biomass; Carotenoids; Rhodotorula; Solvents; Ultrasonic Waves
PubMed: 31669840
DOI: 10.1016/j.ultsonch.2019.104833 -
Frontiers in Bioengineering and... 2020PAL (phenylalanine ammonia lyase) is important for secondary metabolite production in plants and microorganisms. There is broad interest in engineering PAL for its...
PAL (phenylalanine ammonia lyase) is important for secondary metabolite production in plants and microorganisms. There is broad interest in engineering PAL for its biocatalytic applications in industry, agriculture, and medicine. The production of quantities of high-activity enzymes has been explored by gene cloning and heterogeneous expression of the corresponding protein. Here, we cloned the cDNA of PAL (PAL) and introduced codon optimization to improve protein expression in and enzyme activities . The PAL gene was cloned by reverse transcription and named -wt. It had a full-length of 2,121 bp and encoded a 706-amino-acid protein. The -wt was inefficiently expressed in , even when the expression host and physical conditions were optimized. Therefore, codon optimization was used to obtain the corresponding gene sequence, named -opt, in order to encode the same amino acid for the PAL protein. The recombinant protein encoded by -opt, named PAL-opt, was successfully expressed in and then purified to detect its enzymatic activity . Consequently, 55.33 ± 0.88 mg/L of PAL-opt protein with a specific activity of 1,219 ± 147 U/mg and value of 609 μM for substrate L-phenylalanine was easily obtained. The enzyme protein also displayed tyrosine ammonia lyase (TAL)-specific activity of 80 ± 2 U/mg and value of 13.3 μM for substrate L-tyrosine. The bifunctional enzyme PAL/TAL (PAL-opt) and its easy expression advantage will provide an important basis for further applications.
PubMed: 33614604
DOI: 10.3389/fbioe.2020.610506 -
Frontiers in Nutrition 2023Tofu whey wastewater (TWW) is the wastewater of tofu processing, which is rich in a variety of nutrients. can make full use of TWW to ferment and reproduce yeast cells,...
BACKGROUND
Tofu whey wastewater (TWW) is the wastewater of tofu processing, which is rich in a variety of nutrients. can make full use of TWW to ferment and reproduce yeast cells, produce carotenoids and other nutrients, improve the utilization value of TWW, and reduce environmental pollution and resource waste.
METHODS
In this study, the nutrient composition changes of TWW treated by were analyzed to reformulate TWW medium, and the optimal composition and proportion of TWW medium that can improve the biomass and carotenoids production of were explored. Meanwhile, the liquid obtained under these conditions was used to prepare biological feed for laying hens, and the effect of growing on TWW as substrate on laying performance and egg quality of laying hens were verified.
RESULTS
The results showed that the zinc content of TWW after fermentation increased by 62.30%, the phosphorus content decreased by 42.31%, and the contents of vitamin B1, B2 and B6 increased to varying degrees. The optimal fermentation conditions of in the TWW medium were as follow: the initial pH was 6.40, the amount of soybean oil, glucose and zinc ions was 0.80 ml/L, 16.32 g/L, and 20.52 mg/L, respectively. Under this condition, the biomass of reached 2.23 g/L, the carotenoids production was 832.86 μg/g, and the number of effective viable yeast count was 7.08 × 10 cfu/ml. In addition, the laying performance and egg quality of laying hens fed biological feed were improved.
DISCUSSION
In this study, we analyzed the composition changes of TWW, optimized the fermentation conditions of in TWW medium, explored the influence of utilizing TWW on laying layers, and provided a new idea for the efficient utilization of TWW.
PubMed: 36908914
DOI: 10.3389/fnut.2023.1125720 -
Frontiers in Microbiology 2021Diazinon is an organophosphorus pesticide widely used to control cabbage insects, cotton aphids and underground pests. The continuous application of diazinon in... (Review)
Review
Diazinon is an organophosphorus pesticide widely used to control cabbage insects, cotton aphids and underground pests. The continuous application of diazinon in agricultural activities has caused both ecological risk and biological hazards in the environment. Diazinon can be degraded via physical and chemical methods such as photocatalysis, adsorption and advanced oxidation. The microbial degradation of diazinon is found to be more effective than physicochemical methods for its complete clean-up from contaminated soil and water environments. The microbial strains belonging to sp., sp., , , , , and were found to be very promising for the ecofriendly removal of diazinon. The degradation pathways of diazinon and the fate of several metabolites were investigated. In addition, a variety of diazinon-degrading enzymes, such as hydrolase, acid phosphatase, laccase, cytochrome P450, and flavin monooxygenase were also discovered to play a crucial role in the biodegradation of diazinon. However, many unanswered questions still exist regarding the environmental fate and degradation mechanisms of this pesticide. The catalytic mechanisms responsible for enzymatic degradation remain unexplained, and ecotechnological techniques need to be applied to gain a comprehensive understanding of these issues. Hence, this review article provides in-depth information about the impact and toxicity of diazinon in living systems and discusses the developed ecotechnological remedial methods used for the effective biodegradation of diazinon in a contaminated environment.
PubMed: 34790174
DOI: 10.3389/fmicb.2021.717286