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1.
Int J Mol Sci ; 24(2)2023 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-36674962

RESUMO

Yeast acquisition begins at birth; however, the contribution of the mother on yeast transmission to the offspring and associated resistance is yet to be clarified. The aim of this study was to explore the vertical transmission of yeasts and their antifungal susceptibility profile in early life. Oral, fecal, and breastmilk samples were collected from 73 mother-child pairs four to twelve weeks after delivery and cultured on Sabouraud dextrose agar with chloramphenicol. The isolates were identified by MALDI-TOF MS. The vertical transmission was studied by microsatellite genotyping. Antifungal susceptibility was determined for fluconazole, voriconazole, miconazole, anidulafungin, and nystatin by broth microdilution assay, following CLSI-M60 guidelines. A total of 129 isolates were identified from 53% mother-child pairs. We verified the vertical transmission of Candida albicans (n = three mother-child pairs) and Candida parapsilosis (n = one mother-child pair) strains, including an antifungal resistant strain transmitted from breastmilk to the gut of a child. Most isolates were susceptible to the tested antifungals, with the exception of four C. albicans isolates and one R. mucilaginosa isolate. The vertical transmission of yeasts happens in early life. This is the first work that demonstrated the role of the mother as a source of transmission of antifungal-resistant yeasts to the child.


Assuntos
Antifúngicos , Leite Humano , Recém-Nascido , Humanos , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Candida , Leveduras , Boca , Relações Mãe-Filho , Testes de Sensibilidade Microbiana , Farmacorresistência Fúngica
2.
Lett Appl Microbiol ; 76(1)2023 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-36688769

RESUMO

Pichia and Candida species include biofilm-forming yeasts able to spoil foods and beverages. Strains belonging to 10 Pichia and Candida species isolated from apples, grape musts, and wines were analysed. They were subjected to molecular typing and characterized for their ability to grow and ferment must for cider and wine production, and for their biofilm properties. All strains grew similarly in apple and grape must. Glucose-fermenting strains displayed differentiated fermentation performances. Great variation in SO2 and ethanol sensitivity was observed among the strains. Pichia manshurica strains showed high tolerance to both molecules. Eleven and five surface-spreading biofilm (MAT) phenotypes were identified in solid and liquid media, respectively. Strains produced biofilms with variable thicknesses and widths in culture tubes. Cell adherence and aqueous-hydrocarbon biphasic hydrophobicity assays were carried out. Some Pichia manshurica and P. membranifaciens strains exhibited a high capacity to form a thick biofilm and had high cell adherence and hydrophobicity values. These strains could be more likely to colonize the internal surfaces of tanks. This study evidenced that some Pichia and Candida strains can proliferate during apple and grape must fermentation and may be detrimental the beverage quality, due to their specific biofilm properties.


Assuntos
Malus , Vitis , Vinho , Pichia/metabolismo , Candida/metabolismo , Vitis/metabolismo , Leveduras/metabolismo , Vinho/análise , Fermentação
3.
Lett Appl Microbiol ; 76(1)2023 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-36688752

RESUMO

The present study aimed to reveal the microbial (bacteria and yeast) composition of raw milk from dairy camel (n = 10), cow (n = 10) and goat (n = 10) in North-western Nigeria. High-throughput DNA metabarcoding was used to compare microbial compositions in raw milk among the three species. Although the three species had similar dominant bacterial (Firmicutes and Proteobacteria) and yeast (Ascomycota and Basidiomycota) phyla, their microbial compositions at the genus level were noticeably different. The top differentially abundant bacterial and yeast genera (percentage abundance) were Lactobacillus (36%), Streptococcus (34%), Enterococcus (12%), Kluyveromyces (28%), Saccharomyces (24%), and Candida (18%), respectively. Principal coordinate analysis based on unweighted UniFrac values revealed significant differences in the structure of bacterial communities and no differences in yeast communities in milk samples from the three species. This study provides insight into the rich and diverse bacterial and yeast communities in raw animal milk consumed in Nigeria, which could play beneficial roles or pose health threats to consumers. However, further research on the economic significance of the microbial community in animal milk consumed in Nigeria is required.


Assuntos
Bactérias , Leite , Animais , Feminino , Bovinos , Leite/microbiologia , Nigéria , Enterococcus , Streptococcus , Leveduras
4.
Bioresour Technol ; 370: 128573, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36603754

RESUMO

Oleaginous yeasts-derived microbial lipids provide a promising alternative feedstock for the biodiesel industry. However, hyperosmotic stress caused by high sugar concentration during fermentation significantly prevents high cell density and productivity. Isolation of new robust osmophilic oleaginous species from specific environment possibly resolves this issue to some extent. In this study, the cultivable yeast composition of honeycombs was investigated. Totally, 11 species of honeycomb-associated cultivable yeast were identified and characterized. Among them, an osmophilic yeast strain, designated as Rhodotorula toruloides C23 was featured with excellent lipogenic and carotenogenic capacity and remarkable cell growth using glucose, xylose or glycerol as feedstock, with simultaneous production of 24.41 g/L of lipids and 15.50 mg/L of carotenoids from 120 g/L glucose in 6.7-L fermentation. Comparative transcriptomic analysis showed that C23 had evolved a dedicated molecular regulation mechanism to maintain their high simultaneous accumulation of intracellular lipids and carotenoids and cell growth under high sugar concentration.


Assuntos
Lipídeos , Rhodotorula , Leveduras , Rhodotorula/genética , Carotenoides , Glucose
5.
World J Microbiol Biotechnol ; 39(3): 81, 2023 Jan 17.
Artigo em Inglês | MEDLINE | ID: mdl-36646798

RESUMO

Hexavalent chromium is resistant to degradation and harmful toxic substance to environment and community health. Physicochemical treatment methods are demanding high cost, used large quantities of chemicals & energy, release large amount of secondary toxic degradants. Mycoremediation is an eco-friendly alternative treatment method. The main objective of this research is to isolate and characterize chrome (VI) tolerant fungi from farm soil & industry effluent for mycoremedation purpose. The screening and isolation of yeast was carried out on potato dextrose agar media. PDA and broth assay test for fungi tolerance to hexavalent chromium at different concentration, temperature and pH was evaluated. Fungi species was identified biochemically using Biolog Microstation depending on carbon utilization and chemical sensitivity test. The result revealed that 10 yeast species was identified with full ID from effluent waste and farm soil based on their probability ≥  75% and similarity index ≥  0.5 as well as their Cr (VI) tolerance ability up to 2500 ppm. These are Yarrowia lipolytica (100%, 0.7), Cryptococcus luteolus(100%, 0.64), Rhodotorula aurantiaca A(100%, 0.62), Ustilago maydis(100%, 0.58) Trichosporon beigelii B (100%, 0.51), Cryptococcus terreus A (100%, 0.62), Zygosaccharomyces bailii (98%, 0.65), Nadsoniafulvenscens (90%, 0.62), Schizoblastosporonstarkeyihenricii (89%, 0.56), Endomycopsis vivi (84%, 0.62), Rhodotorula pustula (Sim, 0.59). Two yeast species Yarrowia lipolytica and Nadsoniafulvenscens show the highest growth mean Optical density (OD) measure 0.74 ± 0.2 & 0.60 ± 0.2 respectively at pH 7 & 25 °C. The highest tolerance index (mm) was recorded by Schizoblastosporon starkey henricii 0.3067 ± 0.152. Cr (VI)-tolerance ability of these yeast strains used in the development of chromium-bioremediation technologies provide an alternative option for chromium sequestration after HPLC analysis& molecular characterization.


Assuntos
Solo , Verduras , Etiópia , Fazendas , Cromo/metabolismo , Leveduras/metabolismo , Biodegradação Ambiental
6.
Food Microbiol ; 111: 104195, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-36681399

RESUMO

Understanding bacteria and yeasts communities can reduce unpredictable changes of apple cider. In this study, apple juice inoculated with Saccharomyces cerevisiae WET 136 and fermented spontaneously were compared, the relationships of bacteria, yeasts, organic acids, and volatiles were analyzed. Results showed that microbial diversity affected the fermentation, organic acids and volatiles in apple ciders. In the first four spontaneous fermentation days, LAB (lactic acid bacteria) multiplied and reached 7.89 lg CFU/mL, and then triggered malolactic fermentation (MLF), leading to malic acid decreased by 3880.52 mg/L and lactic acid increased by 4787.55 mg/L. The citric, succinic and fumaric acids content was 2171.14, 701.51 and 8.06 mg/L lower than that in inoculated cider, respectively. Although the yeasts multiplied during spontaneous fermentation, it did not reach 7.50 lg CFU/mL until the 5th day, which led to a long lag period, as well as later and lower production of acetaldehyde and higher alcohols. The inoculated yeast inhibited LAB, acetic acid bacteria, Rahnella, and non-Saccharomyces. Yeasts were the key to produce citric acid, acetaldehyde and 3-methyl-1-butanol in apple cider; while bacteria were closely related to the formation of lactic acid, acetic acid and ethyl acetate. It suggested that low higher alcohols and acetaldehyde can be realized by selecting yeasts, and Leuconostoc pseudomesenteroides can work as candidate to reduce L-malic and citric acids in apple cider.


Assuntos
Malus , Bebidas Alcoólicas/microbiologia , Leveduras , Álcoois , Fermentação , Saccharomyces cerevisiae , Bactérias , Acetaldeído
7.
Int J Mol Sci ; 24(2)2023 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-36674823

RESUMO

Three killer toxins that were previously investigated, one excreted by Kluyveromyces wickerhamii and two by different strains of Wickerhamomyces anomalus, were produced at the pilot scale, lyophilized and characterized, and the formulates were assessed for their zymocidial effect against Brettanomyces bruxellensis spoilage yeast. A comparative analysis allowed the evaluation of the minimum inhibitory concentration (MIC) against a sensitive strain. Fungicidal and fungistatic concentrations were used to evaluate the cytocidal effect using a cytofluorimetric approach that confirmed the lethal effect of all lyophilized formulates against B. bruxellensis spoilage yeasts. Moreover, the potential killer toxins' cytotoxicity against human intestinal cells (Caco-2) were evaluated to exclude any possible negative effect on the consumers. Finally, the effective lethal effect of all three lyophilized killer toxins toward B. bruxellensis sensitive strain were tested. The results indicated that all of them acted without dangerous effects on the human epithelial cells, opening the way for their possible commercial application. In particular, D15 showed the lowest MIC and the highest activity, was evaluated also in wine, revealing a strong reduction of Brettamonyces yeast growth and, at the same time, a control of ethyl phenols production.


Assuntos
Brettanomyces , Toxinas Biológicas , Vinho , Humanos , Células CACO-2 , Leveduras , Toxinas Biológicas/farmacologia , Microbiologia de Alimentos
8.
Genes (Basel) ; 14(1)2023 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-36672901

RESUMO

Psychrophilic yeasts are distributed widely on Earth and have developed adaptation strategies to overcome the effect of low temperatures. They can adapt to low temperatures better than bacteriophyta. However, to date, their whole-genome sequences have been limited to the analysis of single strains of psychrophilic yeasts, which cannot be used to reveal their possible psychrophilic mechanisms to adapt to low temperatures accurately and comprehensively. This study aimed to compare different sources of psychrophilic yeasts at the genomic level and investigate their cold-adaptability mechanisms in a comprehensive manner. Nine genomes of known psychrophilic yeasts and three representative genomes of mesophilic yeasts were collected and annotated. Comparative genomic analysis was performed to compare the differences in their signaling pathways, metabolic regulations, evolution, and psychrophilic genes. The results showed that fatty acid desaturase coding genes are universal and diverse in psychophilic yeasts, and different numbers of these genes exist (delta 6, delta 9, delta 12, and delta 15) in the genomes of various psychrophilic yeasts. Therefore, they can synthesize polyunsaturated fatty acids (PUFAs) in a variety of ways and may be able to enhance the fluidity of cell membranes at low temperatures by synthesizing C18:3 or C18:4 PUFAs, thereby ensuring their ability to adapt to low-temperature environments. However, mesophilic yeasts have lost most of these genes. In this study, psychrophilic yeasts could adapt to low temperatures primarily by synthesizing PUFAs and diverse antifreeze proteins. A comparison of more psychrophilic yeasts' genomes will be useful for the study of their psychrophilic mechanisms, given the presence of additional potential psychrophilic-related genes in the genomes of psychrophilic yeasts. This study provides a reference for the study of the psychrophilic mechanisms of psychrophilic yeasts.


Assuntos
Temperatura Baixa , Leveduras , Leveduras/genética , Ácidos Graxos Insaturados , Adaptação Fisiológica/genética , Aclimatação/genética
9.
Bioresour Technol ; 370: 128527, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36572157

RESUMO

Single-cell protein (SCP) from potato starch processing wastewater (PSPW) shows great potential against protein scarcity and unsustainable production of plant and animal proteins. In this study, five yeasts were selected to conduct a series of PSPW fermentation for obtaining high-value SCP by optimizing fermentation conditions. The yeast combination was optimized as Candida utilis, Geotrichum candidum and Candida tropicalis with the volume proportion of 9:5:1. The inoculum size, temperature, rotation speed and initial pH were optimized at 12 %, 24℃, 200 r·min-1 and âˆ¼ 4.13 (natural pH), respectively. At the optimal conditions, SCP yield of 3.06 g·L-1 and water-soluble protein of 17.32 % were obtained with the chemical oxygen demand removal of 56.9 %. A resource-recycling process of PSPW was proposed by coupling yeast fermentation and up-flow anaerobic sludge blanket (UASB) treatment to achieve simultaneous high-level organic removal and SCP production, which could be a promising alternative technology for PSPW treatment.


Assuntos
Amido , Leveduras , Fermentação , Esgotos , Reatores Biológicos , Eliminação de Resíduos Líquidos
10.
Appl Microbiol Biotechnol ; 107(2-3): 915-929, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36576569

RESUMO

BACKGROUND: Monitoring jar fermenter-cultured microorganisms in real time is important for controlling productivity of bioproducts in large-scale cultivation settings. Morphological data is used to understand the growth and fermentation states of these microorganisms during monitoring. Oleaginous yeasts are used for their high productivity of single-cell oils but the relationship between lipid productivity and morphology has not been elucidated in these organisms. RESULTS: In this study, we investigated the relationship between the morphology of oleaginous yeasts (Lipomyces starkeyi and Rhodosporidium toruloides were used) and their cultivation state in a large-scale cultivation setting using a real-time monitoring system. We combined this with deep learning by feeding a large amount of high-definition cell images obtained from the monitoring system to a deep learning algorithm. Our results showed that the cell images could be grouped into 7 distinct groups and that a strong correlation existed between each group and its biochemical activity (growth and oil-productivity). CONCLUSIONS: This is the first report describing the morphological variations of oleaginous yeasts in a large-scale cultivation, and describes a promising new avenue for improving productivity of microorganisms in large-scale cultivation through the use of a real-time monitoring system combined with deep learning. KEY POINTS: • A real-time monitoring system followed the morphological change of oleaginous yeasts. • Deep learning grouped them into 7 distinct groups based on their morphology. • A correlation between the cultivation state and the shape of the yeast was observed.


Assuntos
Aprendizado Profundo , Leveduras , Óleos , Fermentação , Reatores Biológicos
11.
Methods Mol Biol ; 2588: 131-156, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36418686

RESUMO

Colonization of surfaces in the human body by microorganisms is an early, essential, step in the initiation of infectious disease. We have developed in vitro assays to investigate interactions between yeast or bacterial cells and human tissues, fluids, or prostheses. Such assays can be used to identify the adhesins, ligands, and receptors involved in these interactions, for example, by determining which components of the microbe or human tissue/fluid interfere with adherence in the assay. The assays can also be applied to find ways of preventing adhesion, and subsequent disease, by investigating the effects of different conditions and added compounds on adherence in the in vitro assays. Here we describe assays for measuring adhesion of the oral yeast Candida albicans, a common commensal and opportunistic pathogen, or the bacterium Staphylococcus epidermidis, which is not normally pathogenic but is known to form biofilms on medical prostheses. The assays described belong to two approaches to investigating adhesion and biofilm formation: (i) retention at a fixed time point following liquid washes, and (ii) retention against a continuous flow of medium.


Assuntos
Candida albicans , Leveduras , Humanos , Biofilmes , Staphylococcus epidermidis , Adesinas Bacterianas
12.
Food Chem ; 407: 134810, 2023 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-36565578

RESUMO

Brewing produces significant amounts of solid waste during the process: spent cereals (BSG), hops and spent yeast (BSY). These residues are sustainable sources of valuable nutrients and functional compounds like proteins, polyphenols, and polysaccharides. This review describes the three solid wastes and the different extraction techniques for protein recovery. The protein obtained can be used as a new source of non-animal protein or as a functional and bioactive ingredient. Particular attention was given to methods using conventional technologies (alkaline and ethanolic extraction) and more innovative approaches (enzymes, microwaves, ultrasound, pressurized liquids and sub-critical water extraction). Although the BSG is used in some industrial applications, studies in operating conditions, cost, energy efficiency, and product performance are still required to consolidate these solid wastes as a source of non-animal protein. The application of proteins is also an important question when choosing the extraction method.


Assuntos
Grão Comestível , Resíduos Sólidos , Resíduos Sólidos/análise , Grão Comestível/química , Polifenóis/análise , Leveduras , Água/análise
13.
Fungal Genet Biol ; 164: 103767, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36529368

RESUMO

Autochthonous yeasts of oenological origin are adapted to highly stressful and selective environments, which makes them potential candidates for probiotics. The objective of the present study was to explore the probiotic potential of 96 native yeasts of oenological origin, their biosafety, resistance to gastrointestinal tract conditions and adhesion properties. Regarding biosafety, 66 isolates shown negative hemolytic activity, negative urease activity and susceptibility to 3 or more antifungals. After the gastrointestinal resistance test, 15 isolates were selected that showed growth at different temperatures, tolerance to low pH and the presence of bile salts in in vitro tests. In general, survival after simulated conditions of the gastrointestinal tract was high and more restrictive was the duodenal. The results of the adhesion properties showed highly variable hydrophobicity and a high percentage of autoaggregation at 24 h. The maximum production of biofilm was detected in the Pichia strains. Of a total of 96 yeast strains, 15 non-Saccharomyces yeasts presented suitable properties as probiotic candidates. The native winemaking strains performed better than the reference probiotic strain, Saccharomyces cerevisiae var. boulardii CNCM I-745, which reaffirms that these strains are promising probiotic candidates and further studies are necessary to confirm their probiosis.


Assuntos
Probióticos , Vinho , Bioprospecção , Leveduras/genética , Saccharomyces cerevisiae , Trato Gastrointestinal
14.
J Food Sci ; 88(1): 462-476, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36529878

RESUMO

Recently, the use and commercial availability of non-Saccharomyces yeasts (NSY) in winemaking to reduce alcohol content have increased. However, research exploring the influence on sensory quality of the wine, particularly during storage, is limited. Therefore, the objective of this study was to characterize the sensory profiles of Merlot and Chardonnay wines made with pectinase-producing NSY, with added substrate, that is, pectin. Apple pectin (0 or 0.5 g/L) was added to Merlot and Chardonnay grape musts after inoculation with (a) only Saccharomyces cerevisiae or (b) a three species mixture of NSY; after 3 days, S. cerevisiae was added. Addition of NSY with added pectin resulted in higher concentrations of d-galacturonic acid and glycerol concentration in the wines after 6 months of aging. However, mouthfeel (viscosity or weight) of wines with or without added pectin as determined by a sensory evaluation panel was not altered by the presence of these yeasts. Significant interactions among the yeast utilized, pectin addition, and 6-month aging affected some flavors (solvent) of Merlot, while addition of NSY increased other attributes (cherry) during aging. No sensory differences were perceived among Chardonnay samples due to NSY; however, aging from 6 to 18 months increased the intensity of 40 sensory attributes. Though mouthfeel was not specifically affected, the utilization of NSY may be a useful tool to alter wine quality in Merlot by increasing specific aromas during storage. PRACTICAL APPLICATION: We found that must fermented with pectinase-producing non-Saccharomyces yeasts (NSY) modified the chemical composition of the final young wine. After one additional year of aging, an increase in cherry flavor was observed in Merlot wines made with NSY, which may increase perceived quality. Thus, the use of these pectinase-producing NSY may be a useful tool for winemakers.


Assuntos
Saccharomyces , Vitis , Vinho , Vinho/análise , Saccharomyces cerevisiae , Poligalacturonase , Fermentação , Leveduras , Pectinas
15.
Food Chem ; 408: 135121, 2023 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-36521294

RESUMO

Brazil is the largest producer of oranges worldwide, as well as one of the largest producers of orange juice. Alcoholic fermented beverages have been considered a marketable alternative for oranges. In this study, four S. cerevisiae commercial yeasts were evaluated for metabolites generated during orange juice (cv. 'Pêra D9') fermentation. Alcohols, sugars, and organic acids were evaluated by HPLC-DAD-RID during fermentation, and phenolic compounds were analyzed in fermented orange. Orange juice and fermented oranges were also subjected to digestion simulations. The yeasts presented an adequate fermentation activity, based on sugar consumption, and high ethanol (>10.5%) and glycerol (4.8-5.5 g/L) contents. The yeast strains T-58 and US-05 produced high levels of lactic acid. Phenolic compounds and antioxidant activity did not differ amongst yeasts, presenting hesperidin levels between 115 and 127 mg/L, respectively. The fermented orange showed a >70% bioaccessibility, compared to juice, especially for catechin, epigallocatechin-gallate, procyanidin-B2, rutin, and procyanidin-B1.


Assuntos
Citrus sinensis , Proantocianidinas , Saccharomyces cerevisiae/metabolismo , Proantocianidinas/metabolismo , Fermentação , Citrus sinensis/metabolismo , Brasil , Fenóis/análise , Leveduras/metabolismo , Etanol/metabolismo
16.
Biotechnol Lett ; 45(2): 263-272, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36586052

RESUMO

The yeast Cyberlindnera xylosilytica UFMG-CM-Y309 has been identified as a promising new xylitol producer from sugarcane bagasse hemicellulosic hydrolysate (SCHH). However, SCHH pretreatment process generates byproducts, which are toxic to cell metabolism, including furans, phenolic compounds, and carboxylic acids, such as acetic acid, typically released at high concentrations. This research aims to reduce acetic acid in sugarcane hemicellulose hydrolysate concomitantly with xylitol production by yeast strain Cy. xylosilytica UFMG-CM-Y309 in a bioreactor by strategically evaluating the influence of volumetric oxygen transfer coefficient (kLa) (21 and 35 h-1). Experiments were conducted on a bench bioreactor (2 L volumetric capacity) at different initial kLa values (21 and 35 h-1). SCHH medium was supplemented with rice bran extract (10 g L-1) and yeast extract (1 g L-1). Cy. xylosilytica showed high xylitol production performance (19.56 g L-1), xylitol yield (0.56 g g-1) and, maximum xylitol-specific production rate (µpmáx 0.20 gxylitol·g-1 h-1) at kLa value of 21 h-1, concomitantly slowing the rate of acetic acid consumption. A faster acetic acid consumption (100%) by Cy. xylosilytica was observed at kLa of 35 h-1, concomitantly with an increase in maximum cellular growth (14.60 g L-1) and reduction in maximum xylitol production (14.56 g L-1 and Yp/s 0.34 g g-1). This study contributes to pioneering research regarding this yeast performance in bioreactors, emphasizing culture medium detoxification and xylitol production.


Assuntos
Celulose , Saccharum , Celulose/metabolismo , Xilitol , Ácido Acético/metabolismo , Hidrólise , Reatores Biológicos , Leveduras/metabolismo , Fermentação
17.
Sensors (Basel) ; 22(23)2022 Nov 27.
Artigo em Inglês | MEDLINE | ID: mdl-36501924

RESUMO

Advanced Therapy Medicinal Products are promising drugs for patients in therapeutic impasses. Their complex fabrication process implies regular quality controls to monitor cell concentration. Among the different methods available, optical techniques offer several advantages. Our study aims to measure cell concentration in real time in a potential closed-loop environment using white light spectroscopy and to test the possibility of simultaneously measuring concentrations of several species. By analyzing the shapes of the absorption spectra, this system allowed the quantification of T-cells with an accuracy of about 3% during 30 h of cultivation monitoring and 26 h of doubling time, coherent with what is expected for normal cell culture. Moreover, our system permitted concentration measurements for two species in reconstructed co-cultures of T-cells and Candida albicans yeasts. This method can now be applied to any single or co-culture, it allows real-time monitoring, and can be easily integrated into a closed system.


Assuntos
Candida albicans , Linfócitos T , Humanos , Técnicas de Cocultura , Leveduras , Técnicas de Cultura de Células
18.
BMC Bioinformatics ; 23(1): 555, 2022 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-36544090

RESUMO

MOTIVATION: In eukaryotes, homologous recombination between the parental genomes frequently occurs during the evolutionary conserved process of meiosis, generating the genetic diversity transmitted by the gametes. The genome-wide determination of the frequency and location of the recombination events can now be efficiently performed by genotyping the offspring's polymorphic markers. However, genotyping recombination in complex hybrid genomes with existing methods remains challenging because of their strain and ploidy specificity and the degree of diversity and complexity of the parental genomes, especially in [Formula: see text] polyploids. RESULTS: We present UGDR, a pipeline to genotype the polymorphisms of complex hybrid yeast genomes. It is based on optimal mapping strategies of NGS reads, comparative analyses of the allelic ratio variation and read depth coverage. We tested the UGDR pipeline with sequencing reads from recombined hybrid diploid yeast strains and various clinical strains exhibiting different degrees of ploidy. UGDR allows to plot the markers distribution and recombination profile per chromosome. CONCLUSION: UGDR detects and plots recombination events in haploids and polyploid yeasts, which facilitates the discovery and understanding of the yeast genetic recombination map and identify new out-performing recombinants.


Assuntos
Genoma , Poliploidia , Ploidias , Diploide , Polimorfismo Genético , Leveduras
19.
PLoS Biol ; 20(12): e3001877, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36520709

RESUMO

The yeast Komagataella phaffii (formerly called Pichia pastoris) is used widely as a host for secretion of heterologous proteins, but only a few isolates of this species exist and all the commonly used expression systems are derived from a single genetic background, CBS7435 (NRRL Y-11430). We hypothesized that other genetic backgrounds could harbor variants that affect yields of secreted proteins. We crossed CBS7435 with 2 other K. phaffii isolates and mapped quantitative trait loci (QTLs) for secretion of a heterologous protein, ß-glucosidase, by sequencing individual segregant genomes. A major QTL mapped to a frameshift mutation in the mannosyltransferase gene HOC1, which gives CBS7435 a weaker cell wall and higher protein secretion than the other isolates. Inactivation of HOC1 in the other isolates doubled ß-glucosidase secretion. A second QTL mapped to an amino acid substitution in IRA1 that tripled ß-glucosidase secretion in 1-week batch cultures but reduced cell viability, and its effects are specific to this heterologous protein. Our results demonstrate that QTL analysis is a powerful method for dissecting the basis of biotechnological traits in nonconventional yeasts, and a route to improving their industrial performance.


Assuntos
Celulases , Saccharomycetales , Pichia/genética , Pichia/metabolismo , Saccharomycetales/genética , Leveduras , Celulases/metabolismo , Proteínas Recombinantes/metabolismo
20.
PLoS Biol ; 20(12): e3001911, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36525401

RESUMO

Improvements to the production of proteins in industrial yeast species have largely relied on generating variation in a single genetic background. A new study in PLOS Biology leverages natural genetic variation to identify genes and variants with the potential to improve protein yield.


Assuntos
Pichia , Saccharomycetales , Humanos , Pichia/genética , Pichia/metabolismo , Saccharomycetales/genética , Leveduras , Biotecnologia
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