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1.
Methods Mol Biol ; 2477: 149-175, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35524117

RESUMO

Chromatin immunoprecipitation followed by mass spectrometry (ChIP-MS) is a powerful method to identify protein interactions, and has long been used to gain insights into regulatory networks in relevant fungal species as well as many other organisms. In this chapter, we discuss a similar technique called ChIP-SICAP (chromatin immunoprecipitation with selective isolation of chromatin-associated proteins) that overcomes many of the traditional limitations of ChIP-MS, and describe a protocol that allows ChIP-SICAP to be applied to Candida albicans and other yeasts. Notably, the technique design permits stringent washing to remove contaminating proteins and antibodies before subsequent mass spectrometry processing, allows for genome-wide mapping of the bait protein by ChIP-seq after ChIP-SICAP from the same sample through a DNA recovery process, and specifically purifies and identifies proteins associating with chromatin. In the future, ChIP-SICAP will provide the yeast genomics research community an additional method to explore the complex dynamics of the gene-regulatory networks modulating morphology, metabolism and response to stress.


Assuntos
Candida albicans , Leveduras , Candida albicans/genética , Cromatina/genética , Imunoprecipitação da Cromatina/métodos , Redes Reguladoras de Genes , Leveduras/genética
2.
Int J Food Microbiol ; 370: 109652, 2022 Jun 02.
Artigo em Inglês | MEDLINE | ID: mdl-35390573

RESUMO

Yeast plays an important role in the formation of desirable aroma during soy sauce fermentation. In this study, the structure and diversity of yeast communities in seven different soy sauce residues were investigated by ITS sequencing analysis, and then the aroma characteristics of selected yeast species were examined by a combination of gas chromatography-mass spectrometry (GC-MS), headspace solid-phase microextraction (SPME) and liquid-liquid extraction (LLE). A total of 18 yeast genera were identified in seven soy sauce residues. Among them, Candid and Zygosaccharomyces were detected in all samples, followed by Millerozyma, Wickerhamiella, Meyerozyma, Trichosporon and Wickerhamomyces, which were found in more than two-thirds of the samples. Subsequently, eight representative species, isolated from soy sauce residues, were subjected to environmental stress tolerance tests and aroma production tests. Among them, three isolated species were regarded as potential aroma-enhancing microbes in soy sauce. Wickerhamiella versatilis could increase the contents of ethyl ester compounds and alcohols, thereby improving the fruity and alcoholic aroma of soy sauce. Candida sorbosivorans enhanced sweet and caramel-like aroma of soy sauce by producing 4-hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF) and 3-hydroxy-2-methyl-4h-pyran-4-one (maltol). Starmerella etchellsii could enhance the contents of 2,6-dimethylpyrazine, methyl pyrazine and benzeneacetaldehyde. This study is of great significance for the development and application of flavor functional yeasts in soy sauce fermentation.


Assuntos
Alimentos de Soja , Zygosaccharomyces , Fermentação , Odorantes/análise , Alimentos de Soja/análise , Leveduras
3.
Probiotics Antimicrob Proteins ; 14(3): 595-601, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35451745

RESUMO

Mycocins are substances that have the potential to affect other sensitive yeasts or microorganisms. Wickerhamomyces anomalus is a yeast that produces mycocins that have great biotechnological potential, being highly competitive in many habitats, as it is adaptable to a wide range of environmental conditions. Thus, they are targets for studies in different areas, including the environment, industry, and medical sciences. Yeasts of the genus Candida are of great importance due to the high frequency with which they colonize and infect the human host. Yeast infections are often difficult to treat due to the acquisition of resistance against antifungals, leading to studies focusing in new treatment alternatives. This work aims to verify the inhibition of Candida albicans isolated from vaginal secretion by mycocins produced by Wickerhamomyces anomalus. Tests were carried out in solid medium and microdilution tests, where mycocins proved to be efficient in inhibiting the growth of C. albicans, hemolysis, and irritation in an organotypic model, which showed that the mycocins produced by W. anomalus are safe and non-irritating. Thus, the results of this work can provide scientific evidence for the application of mycocins in the production of new antifungal alternatives.


Assuntos
Candida albicans , Saccharomycetales , Antifúngicos/farmacologia , Candida , Feminino , Humanos , Leveduras
4.
Arch Microbiol ; 204(5): 284, 2022 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-35476294

RESUMO

Yeasts isolated from the worker caste of the Colombian leaf-cutting ant, Atta cephalotes (Hymenoptera: Myrmicinae) were cultured and identified by molecular methods. Abundant, persistent, and omnipresent species were classified as "prevalent". Experimental data were compared with information gathered from published reports on the yeast species composition in other leaf-cutting ant species. Diversity analysis was conducted using diversity values (q0, q1, and q2) to compare the richness and abundance of yeasts present in different leaf-cutting ant species. Clustering analysis was carried out to assess the similarity of yeast community according to ant species. The yeast species composition was highly variable among the ant species. A. laevigata and A. capiguara showed the highest degree of similarity and differed from the group composed by A. cephalotes, A. sexdens, A. sexdens rubropilosa, and A. texana. The isolation of dominant yeasts in different ant castes within the different compartments of a colony strongly suggests that the identified microorganisms are not transient but are native to the soil surrounding ant colonies and the substrates used by the ants to grow their fungal cultivars. It is apparent that the ant-fungus mutualism does not operate in an environment devoid of other microbes, but rather that the association must be seen within the context of a background of other microorganisms, particularly the dominant yeasts.


Assuntos
Formigas , Animais , Formigas/microbiologia , Colômbia , Classe Social , Simbiose , Leveduras/genética
5.
Plant Sci ; 319: 111243, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35487651

RESUMO

Cyperus esculentus is considered one of the most promising oil crops due to its oil-rich tuber, wide adaptability and large biomass production. Preferable triacylglycerol (TAG) composition, especially high oleic acid content, makes tuber oil suitable for human consumption and biodiesel production. However, the mechanism underlying oleic acid enrichment in the tuber remains unknown. Plastidial stearoyl-ACP desaturase (SAD) catalyses the formation of monounsaturated fatty acids (MUFAs), which may function crucially for high accumulation of oleic acid in C. esculentus tubers. In this study, two full-length cDNAs encoding SAD were isolated from the developing tubers of C. esculentus, namely, CeSAD1 and CeSAD2, with ORFs of 1194 bp and 1161 bp, respectively. Quantitative RT-PCR analysis showed that CeSAD genes were highly expressed in tubers. The expression pattern during tuber formation was also significantly correlated with fatty acid and oil accumulation dynamics. Overexpression of each CeSAD gene could restore the normal growth of the defective yeast BY4389, indicating that both CeSADs had fatty acid desaturase activity to catalyse MUFA biosynthesis. A tobacco genetic transformation assay demonstrated that both CeSAD enzymes had high enzyme activity. Exogenous addition of exogenous fatty acids to feed yeast revealed that CeSAD1 has a more substantial substrate preference ratio for C18:0 than CeSAD2 did. Moreover, the overexpression of CeSAD1 significantly increased host tolerance against low-temperature stress. Our data add new insights into the deep elucidation of oleic acid-enriched oils in Cyperus esculentus tubers, showing CeSAD, especially CeSAD1, as the target gene in genetic modification to increase oil and oleic yields in oil crops as well as stress tolerance.


Assuntos
Cyperus , Ácidos Graxos Dessaturases , Cyperus/genética , Cyperus/metabolismo , Ácidos Graxos Dessaturases/genética , Ácidos Graxos Dessaturases/metabolismo , Ácidos Graxos/metabolismo , Óleos/metabolismo , Ácido Oleico/metabolismo , Leveduras/metabolismo
6.
World J Microbiol Biotechnol ; 38(6): 93, 2022 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-35441962

RESUMO

As essential structural molecules of fungal cell membrane, ergosterol is not only an important component of fungal growth and stress-resistance but also a key precursor for manufacturing steroid drugs of pharmaceutical or agricultural significance. So far, ergosterol biosynthesis in yeast has been elucidated elaborately, and efforts have been made to increase ergosterol production through regulation of ergosterol metabolism and storage. Furthermore, the same intermediates shared by yeasts and animals or plants make the construction of heterologous sterol pathways in yeast a promising approach to synthesize valuable steroids, such as phytosteroids and animal steroid hormones. During these challenging processes, several obstacles have arisen and been combated with great endeavors. This paper reviews recent research progress of yeast metabolic engineering for improving the production of ergosterol and heterologous steroids. The remaining tactics are also discussed.


Assuntos
Ergosterol , Saccharomyces cerevisiae , Ergosterol/metabolismo , Engenharia Metabólica , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Esteróis , Leveduras/genética , Leveduras/metabolismo
7.
Microb Cell Fact ; 21(1): 70, 2022 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-35468837

RESUMO

BACKGROUND: The yeast genus Komagataella currently consists of seven methylotrophic species isolated from tree environments. Well-characterized strains of K. phaffii and K. pastoris are important hosts for biotechnological applications, but the potential of other species from the genus remains largely unexplored. In this study, we characterized 25 natural isolates from all seven described Komagataella species to identify interesting traits and provide a comprehensive overview of the genotypic and phenotypic diversity available within this genus. RESULTS: Growth tests on different carbon sources and in the presence of stressors at two different temperatures allowed us to identify strains with differences in tolerance to high pH, high temperature, and growth on xylose. As Komagataella species are generally not considered xylose-utilizing yeasts, xylose assimilation was characterized in detail. Growth assays, enzyme activity measurements and 13C labeling confirmed the ability of K. phaffii to utilize D-xylose via the oxidoreductase pathway. In addition, we performed long-read whole-genome sequencing to generate genome assemblies of all Komagataella species type strains and additional K. phaffii and K. pastoris isolates for comparative analysis. All sequenced genomes have a similar size and share 83-99% average sequence identity. Genome structure analysis showed that K. pastoris and K. ulmi share the same rearrangements in difference to K. phaffii, while the genome structure of K. kurtzmanii is similar to K. phaffii. The genomes of the other, more distant species showed a larger number of structural differences. Moreover, we used the newly assembled genomes to identify putative orthologs of important xylose-related genes in the different Komagataella species. CONCLUSIONS: By characterizing the phenotypes of 25 natural Komagataella isolates, we could identify strains with improved growth on different relevant carbon sources and stress conditions. Our data on the phenotypic and genotypic diversity will provide the basis for the use of so-far neglected Komagataella strains with interesting characteristics and the elucidation of the genetic determinants of improved growth and stress tolerance for targeted strain improvement.


Assuntos
Saccharomycetales , Xilose , Carbono/metabolismo , Fenótipo , Pichia/metabolismo , Saccharomycetales/genética , Xilose/metabolismo , Leveduras
8.
Food Res Int ; 155: 111069, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35400447

RESUMO

Nyons table olives, named after the French city where they are processed, are naturally fermented black table olives. Their specificity relies on the use of the "Tanche" olive variety harvested at full maturity and their slow spontaneous fermentation in 10% salt brine driven by yeast populations. This study aimed at investigating the benefit of inoculating autochthonous consortia to produce Nyons table olives by fermentation in 10% salt brine and in reduced salt conditions (8%). Two strategies were evaluated: inoculation with a defined autochthonous consortium and inoculation by spent brine backslopping. To define the consortium, yeasts were selected among 48 autochthonous isolates and key features included high halotolerance, low pectinolytic and proteolytic activities, however none had ß-glucosidase activities. The consortium included eight yeast strains with distinct technological properties belonging to five dominant species, i.e. Citeromyces nyonsensis, Pichia membranifaciens, Wickerhamomyces anomalus, Zygotorulaspora mrakii and Candida atlantica. Fermentation trials were conducted over a year and compared by evaluating microbial community shifts (16S and ITS metagenetics) and volatile profiles (GC-MS). Regarding fermentations with the defined consortium, four out of five species implanted in early stages while one, Pichia membranifaciens, persisted and largely dominated by the end of the fermentation. Altogether, inoculation with the defined consortium did not disrupt microbial shifts compared to traditional fermentations although minor differences were observed in volatile profiles. The backslopping method yielded the highest impact on microbial populations and olive volatile profiles, with higher ester abundances at the end of fermentation. Finally, reduced salt in brine gave very promising results as no deleterious effects on microbial communities, volatile dynamics but also safety criteria of the olives were observed compared to traditional fermented olives.


Assuntos
Olea , Fermentação , Microbiologia de Alimentos , Pichia , Sais , Cloreto de Sódio , Leveduras
9.
An Acad Bras Cienc ; 94(suppl 1): e20210592, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35384975

RESUMO

Antarctic harsh conditions favor the development of microbial adaptations. In this study, a molecular approach was applied to identify/refine the taxonomy of five yeasts isolated from different Antarctic samples, which were tested against ranges of temperature, UV radiations, salinity, and pH. Based on sequencing and phylogenetic analysis, strain CRM 1839 was confirmed as Naganishia sp., and strains CRM 1874, CRM 1565, CRM 2571, and CRM 2576 were identified as Goffeauzyma gilvescens, Goffeauzyma gastrica, Candida atlantica, and Camptobasidium sp., respectively, being this last one possibly a new species. Growth at different temperatures indicates that these yeasts are psychrotolerant, with the exception of Camptobasidium sp., which presents psychrophilic characteristics. G. gastrica recovered from marine sediment showed the best results of resistance to UV radiation, being able to grow even after the exposure to UVB dose of 9144 J/m² and UVC dose of 6102 J/m². C. atlantica isolated from glacier soil showed high cellular growth from 3 to 10% NaCl. The majority of the strains produced higher biomass at pH 7; nevertheless, G. gilvescens showed higher biomass production at pH 9. The studied Antarctic-derived yeasts have adaptations to extreme conditions, which makes them useful for biotechnological applications and studies of extremophiles.


Assuntos
Sedimentos Geológicos , Leveduras , Regiões Antárticas , Filogenia
10.
Food Chem ; 385: 132719, 2022 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-35318172

RESUMO

Plant bioactive compounds have been studied mainly for their beneficial antioxidant properties. Kombucha is a fermented beverage traditionally obtained from fermentation of sweetened black or green tea by a characteristic consortium of yeasts and bacteria. The beverage naturally contains bioactive compounds from teas and their synthesis can be increased during fermentation. This review aims to explore the different bioactive compounds found in kombucha from different substrates, as well as the factors that influence on their synthesis and their amount in the final product. The results suggest phenolic compounds are the main bioactive compounds in kombucha. The substrate type contributes the most to increasing the content of bioactive compounds in the final product; fermentation time and type of sugar also increase the amount of these compounds. Further research suggestions include the combination of strategies to increase bioactive compounds in kombucha, quantification and characterization of the isolated compounds.


Assuntos
Bebidas , Chá , Antioxidantes/análise , Bebidas/análise , Fermentação , Chá/química , Leveduras
11.
Genes (Basel) ; 13(3)2022 02 28.
Artigo em Inglês | MEDLINE | ID: mdl-35327998

RESUMO

Yeasts play important roles in both the environment and in human welfare. While some environmental yeasts positively contribute to nutrient cycling and food production, a significant number of yeast species are opportunistic human pathogens, including several that are tolerant/resistant to commonly used antifungal drugs. At present, most of our understanding of environmental yeasts has come from a few terrestrial environments in selected geographic regions. Relatively little is known about yeast diversity in tropical environments and their potential impacts on human health. Here, we characterize culturable yeasts in 968 environmental samples from eight regions in tropical China. Among the 516 soil, 273 freshwater, and 179 seawater samples, 71.5%, 85.7%, and 43.6% contained yeasts, respectively. A total of 984 yeast isolates were analyzed for their DNA barcode sequences and their susceptibilities to fluconazole. DNA sequence comparisons revealed that the 984 yeast isolates likely belonged to 144 species, including 106 known species and 38 putative novel species. About 38% of the 984 isolates belonged to known human pathogens and the most common species was Candida tropicalis, accounting for 21% (207/984) of all isolates. Further analyses based on multi-locus sequence typing revealed that some of these environmental C. tropicalis shared identical genotypes with clinical isolates previously reported from tropical China and elsewhere. Importantly, 374 of the 984 (38%) yeast isolates showed intermediate susceptibility or resistance to fluconazole. Our results suggest that these environmental yeasts could have significant negative impacts on human health.


Assuntos
Farmacorresistência Fúngica , Fluconazol , Antifúngicos/farmacologia , Farmacorresistência Fúngica/genética , Fluconazol/farmacologia , Humanos , Tipagem de Sequências Multilocus , Leveduras/genética
12.
Am J Bot ; 109(3): 393-405, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35315515

RESUMO

PREMISE: Yeasts are often present in floral nectar and can influence plant fitness directly (independently of pollinators) or indirectly by influencing pollinator visitation and behavior. However, few studies have assessed the effect of nectar yeasts on plant reproductive success or compared effects across different plant species, limiting our understanding of the relative impact of direct vs. indirect effects. METHODS: We inoculated the nectar of six plant species in the field with the cosmopolitan yeast Metschnikowia reukaufii to analyze the direct and indirect effects on female reproductive success over 2 years. The pollinator assemblage for each species was recorded during both flowering years. RESULTS: Direct yeast effects on female fecundity were statistically nonsignificant for all plant species. There were significant indirect, pollinator-mediated effects on fruit production and seed mass for the two species pollinated almost exclusively by bumblebees or hawkmoths, with the direction of the effects differing for the quantity- and quality-related fitness components. There were no consistent effects of the yeast on maternal fecundity for any of the species with diverse pollinator assemblages. CONCLUSIONS: Effects of M. reukaufii on plant reproduction ranged from negative to neutral or positive depending on the plant species. The among-species variation in the indirect effects of nectar yeasts on plant pollination could reflect variation in the pollinator community, the specific microbes colonizing the nectar, and the order of microbial infection (priority effects), determining potential species interactions. Elucidating the nature of these multitrophic plant-pollinator-microbe interactions is important to understand complex processes underlying plant pollination.


Assuntos
Néctar de Plantas , Polinização , Flores , Plantas , Reprodução , Leveduras
13.
Antonie Van Leeuwenhoek ; 115(5): 609-633, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35322327

RESUMO

This report is the first investigation of yeast biodiversity from the oligotrophic hypersaline coastal waters of the Arabian Gulf surrounding Qatar. Yeasts and yeast-like fungi, were cultured from seawater sampled at 13 coastal areas surrounding Qatar over a period of 2 years (December 2013-September 2015). Eight hundred and forty-two isolates belonging to 82 species representing two phyla viz., Ascomycota (23 genera) and Basidiomycota (16 genera) were identified by molecular sequencing. The results indicated that the coastal waters of the Qatari oligotrophic marine environment harbor a diverse pool of yeast species, most of which have been reported from terrestrial, clinical and aquatic sources in various parts of the world. Five species, i.e., Candida albicans, C. parapsilosis, C. tropicalis, Pichia kudriavzevii and Meyerozyma guilliermondii (n = 252/842; 30% isolates) are known as major opportunistic human pathogens. Fifteen species belonging to nine genera (n = 498/842; 59%) and 12 species belonging to seven genera (n = 459/842; 55%) are hydrocarbon degrading yeast and pollution indicator yeast species, respectively. Ascomycetous yeasts were predominant (66.38%; 559/842) as compared to their basidiomycetous counterparts (33.6%; 283/842). The most isolated yeast genera were Candida (28%; 236/842) (e.g., C. aaseri, C. boidinii, C. glabrata, C. intermedia, C. oleophila, C. orthopsilosis, C. palmioleophila, C. parapsilosis, C. pseudointermedia, C. rugopelliculosa, C. sake, C. tropicalis and C. zeylanoides), Rhodotorula (12.7%; 107/842), Naganishia (8.4%; 71/842), Aureobasidium (7.4%; 62/842), Pichia (7.3%; 62/842), and Debaryomyces (6.4%; 54/842). A total of eleven yeast species ( n = 38) isolated in this study are reported for the first time from the marine environment. Chemical testing demonstrated that seven out of the 13 sites had levels of total petroleum hydrocarbons (TPH) ranging from 200 to 900 µg/L, whereas 6 sites showed higher TPH levels (> 1000-21000 µg/L). The results suggest that the yeast community structure and density are impacted by various physico-chemical factors, namely total organic carbon, dissolved organic carbon and sulphur.


Assuntos
Ascomicetos , Basidiomycota , Biodiversidade , Humanos , Catar , Água do Mar/microbiologia , Leveduras
14.
Int J Food Microbiol ; 369: 109627, 2022 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-35305516

RESUMO

Wet coffee fermentation is widely used in coffee-producing regions such as Colombia and Hawaii, but it is not widespread in Brazil. This study aimed to evaluate inoculating the lactic acid bacteria Leuconostoc mesenteroides CCMA1105 and Lactiplantibacillus plantarum CCMA 1065 and the yeasts Saccharomyces cerevisiae CCMA0543 and Torulaspora delbrueckii CCMA0684 as starter cultures on wet coffee fermentation using the SIAF method (self-induced anaerobiosis fermentation). The microbial activity resulted in high consumption of the carbohydrates glucose (98.6%), fructose (97.6%), and sucrose (100%), in addition to the production of lactic and acetic acids, impacting the final quality of the beverage. A total of 108 volatile compounds belonging to 17 classes were identified in the green and roasted coffee samples, including 2,3-butanediol produced by lactic acid bacteria, contributing to coffee's aromatic profile. The final scores for the coffees from the different fermentations ranged from 79.0 to 83.25. The inoculated fermentations were classified as specialty according to the Specialty Coffee Association. Therefore, whole coffee fruit processed via wet using SIAF method and yeast and lactic acid bacteria starter is an alternative for improving wet fermented coffee quality and obtaining coffee beverages with a different sensory profile.


Assuntos
Coffea , Lactobacillales , Torulaspora , Coffea/microbiologia , Café/microbiologia , Fermentação , Leveduras
15.
An Acad Bras Cienc ; 94(suppl 1): e20210540, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35293947

RESUMO

In the Antarctic environment, yeasts are versatile eukaryotes that have shown wide dispersion in different substrates, producing active enzymes in extreme conditions, but their relevance in biotechnological applications is largely unknown. The aim of this study was to evaluate the production of extracellular hydrolases by yeasts isolated from Antarctic lichens and molecularly identify these isolates. From a total of 144 isolates on the screening, 109 (76%) produced at least one of the hydrolases tested, with most activities for proteases 59 (41%), cellulases 58 (40%), esterases 57 (39%), lipases 29 (20%), amylases 23 (16%) and pectinases 20 (14%). Among these isolates, 76 were identified, most belonged to the phylum Basidiomycota (n=73) with the dominance of Vishniacozyma victoriae (n=27), Cystobasidium alpinum (n=3), Mrakia niccombsii (n=3), Cystobasidium laryngis (n=2), Bannozyma yamatoana (n=2), Holtermanniella nyarrowii (n=2), and Glaciozyma martinii (n=2). This study is the first one reporting extracellular enzyme production by yeasts isolated from thallus of the species of Antarctic lichens Lecania brialmontii, Polycauliona candelaria, Usnea capillacea, Cladonia metacorallifera, and Polycauliona regalis. With these data, it's possible to confirm lichens as a source of hydrolase-producing yeasts, reinforcing the potential of these microorganisms in bioprospecting studies of catalytic molecules from polar regions that may be useful in promising biotechnological applications.


Assuntos
Líquens , Hidrólise , Lipase , Peptídeo Hidrolases , Leveduras
16.
Arch. latinoam. nutr ; 72(1): 50-59, mar. 2022. ilus, tab
Artigo em Inglês | LILACS, LIVECS | ID: biblio-1368371

RESUMO

Crohn's disease (CD) is an inflammatory condition that can affect the entire gastrointestinal tract due to an exacerbated and inadequate immune system response. Objective. This study aimed to conduct a systematic review, through clinical trials, about the use of probiotics in humans with CD. Materials and methods. Research was carried out in the PubMed, Scopus and Science Direct databases using the keywords "Crohn's disease" and "probiotics". We conducted the review by searching clinical trials published from 2000 to December 2019. Results. Of 2,164 articles found, only nine were considered eligible for this review. The studies investigated patients with CD at different stages of the pathology, and in three studies the potential effect of probiotics in the active phase was observed; in two, in the remission phase; and in four, after intestinal surgery. The sample size of the studies ranged from 11 to 165 individuals and the age of the participants between 5 and 71 years. Gram-positive bacteria were used in six clinical interventions and in two studies yeasts were used. As for the significant results obtained with the treatment with probiotics, in one study there was beneficial clinical effects in patients and, in another, there was an improvement in intestinal permeability. Conclusion. Currently, it is not possible to establish a recommendation for probiotic therapy to control CD due to the few clinical trials with significant results. There is a need for more research on clinical intervention with probiotics in CD to clarify the action, define doses and time of use(AU)


La enfermedad de Crohn (EC) es una afección inflamatoria que puede afectar todo el tracto gastrointestinal debido a una respuesta del sistema inmunitario exacerbada e inadecuada. Objetivo. Realizar una revisión sistemática, a través de ensayos clínicos, sobre el uso de probióticos en humanos con EC. Materiales y métodos. La investigación se realizó en las bases de datos PubMed, Scopus y Science Direct utilizando las palabras clave "enfermedad de Crohn" y "probióticos". La revisión se hizo en ensayos clínicos publicados desde 2000 hasta diciembre 2019. Resultados. De 2164 artículos encontrados, solo nueve fueron considerados elegibles. Los estudios investigaron pacientes con EC en diferentes etapas de la patología, y en tres estudios se observó el efecto potencial de los probióticos en la fase activa; en dos, en remisión; y en cuatro, tras cirugía intestinal. El tamaño de la muestra fue entre 11 y 165 individuos y la edad entre 5 y 71 años. Se utilizaron bacterias grampositivas en seis intervenciones clínicas y en dos estudios se utilizaron levaduras. En cuanto a los resultados significativos obtenidos con el tratamiento con probióticos, en un estudio hubo efectos clínicos beneficiosos en los pacientes y, en otro, hubo una mejora en la permeabilidad intestinal. Conclusión. Actualmente, no es posible establecer una recomendación de terapia con probióticos para el control de la EC debido a los pocos ensayos clínicos con resultados significativos. Existe la necesidad de más investigación sobre la intervención clínica con probióticos en EC para aclarar la acción, definir dosis y tiempo de uso(AU)


Assuntos
Humanos , Masculino , Feminino , Pré-Escolar , Criança , Adolescente , Adulto , Pessoa de Meia-Idade , Idoso , Doença de Crohn , Probióticos , Trato Gastrointestinal , Bactérias Gram-Positivas , Permeabilidade , Leveduras , Doenças Inflamatórias Intestinais , PubMed , Sistema Imunitário
17.
Nutrients ; 14(3)2022 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-35277029

RESUMO

Kombucha is an increasingly popular functional beverage that has gained attention for its unique combination of phytochemicals, metabolites, and microbes. Previous chemical and microbial composition analyses of kombucha have mainly focused on understanding their changes during fermentation. Very limited information is available regarding nutrient profiles of final kombucha products in the market. In this study, we compared the major chemicals (tea polyphenols, caffeine), antioxidant properties, microbial and metabolomic profiles of nine commercial kombucha products using shotgun metagenomics, internal transcribed spacer sequencing, untargeted metabolomics, and targeted chemical assays. All of the nine kombucha products showed similar acidity but great differences in chemicals, metabolites, microbes, and antioxidant activities. Most kombucha products are dominated by the probiotic Bacillus coagulans or bacteria capable of fermentation including Lactobacillus nagelii, Gluconacetobacter, Gluconobacter, and Komagataeibacter species. We found that all nine kombuchas also contained varying levels of enteric bacteria including Bacteroides thetaiotamicron, Escherischia coli, Enterococcus faecalis, Bacteroides fragilis, Enterobacter cloacae complex, and Akkermansia muciniphila. The fungal composition of kombucha products was characterized by predominance of fermenting yeast including Brettanomyces species and Cyberlindnera jadinii. Kombucha varied widely in chemical content assessed by global untargeted metabolomics, with metabolomic variation being significantly associated with metagenomic profiles. Variation in tea bases, bacteria/yeast starter cultures, and duration of fermentation may all contribute to the observed large differences in the microbial and chemical profiles of final kombucha products.


Assuntos
Polifenóis , Leveduras , Bactérias/metabolismo , Bebidas/análise , Fermentação , Polifenóis/análise , Leveduras/metabolismo
18.
ACS Synth Biol ; 11(4): 1497-1509, 2022 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-35294186

RESUMO

Gene editing technology plays an extremely significant role in synthetic biology and metabolic engineering. Traditional genetic manipulation methods, such as homologous recombination, however, are inefficient, time-consuming, and barely feasible when disrupting multiple genes simultaneously. Starmerella bombicola, a nonconventional yeast that overproduces sophorolipids, lacks convenient genetic tools for engineering strains. Here, we developed an efficient CRISPR-Cas9 genome editing technology by combining molecular element mining and expression system optimization for S. bombicola. This CRISPR-Cas9 system improved the efficiency of gene-integration/target gene-introducing disruption by homology-directed repair and realized the multi-gene simultaneous disruptions. Based on this CRISPR-Cas9 system, we also further constructed an engineered strain via the in vivo assembly of multiple DNA fragments (10 kb) that can produce acid-type sophorolipids. These results showed that the CRISPR-Cas9 system may be an efficient and convenient strategy to perform genetic manipulation in S. bombicola.


Assuntos
Sistemas CRISPR-Cas , Saccharomycetales , Sistemas CRISPR-Cas/genética , Edição de Genes/métodos , Saccharomycetales/genética , Leveduras/genética
19.
FEMS Yeast Res ; 22(1)2022 03 09.
Artigo em Inglês | MEDLINE | ID: mdl-35157047

RESUMO

The use of non-Saccharomyces yeasts in the winemaking process may have several positive outcomes. Kluyveromyces marxianus has recently been revealed as a promising species for this industry. While the majority of studies mention the use of K. marxianus in various industries including food production (e.g. dairy and cocoa), recent studies have also shown that its aroma and pectinase production make it a suitable yeast for the wine industry. Nevertheless, only particular strain, IWBT Y885, was investigated. In this study, five different K. marxianus strains as well as one protoplast fusant (BF2020) were compared to strain Y885. These comparisons focused on various oenological traits such as fermentation performance, fermentation metabolites, hydrogen sulfide, and pectinase production. Throughout the study, variations were found between the K. marxianus strains investigated. Indeed, although common traits such as high pectinase activity appeared conserved among K. marxianus strains, a fairly large phenotypic diversity was also evident. Using cluster analysis, strain groupings emerged with strains L01, L05, Y885, and BF2020 grouping together. Similarly, strains L02 and L04 grouped together while strain L03 appearing to show the most variation between the strains investigated. Variation between strains was observed regardless of the original source of isolation.


Assuntos
Kluyveromyces , Poligalacturonase , Fermentação , Kluyveromyces/genética , Kluyveromyces/metabolismo , Poligalacturonase/metabolismo , Leveduras/metabolismo
20.
Commun Biol ; 5(1): 144, 2022 02 17.
Artigo em Inglês | MEDLINE | ID: mdl-35177796

RESUMO

Genomic rearrangements often generate phenotypic diversification. We previously reported the TAQing system where genomic rearrangements are induced via conditional activation of a restriction endonuclease in yeast and plant cells to produce mutants with marked phenotypic changes. Here we developed the TAQing2.0 system based on the direct delivery of endonucleases into the cell nucleus by cell-penetrating peptides. Using the optimized procedure, we introduce a heat-reactivatable endonuclease TaqI into an asexual industrial yeast (torula yeast), followed by a transient heat activation of TaqI. TAQing2.0 leads to generation of mutants with altered flocculation and morphological phenotypes, which exhibit changes in chromosomal size. Genome resequencing suggested that torula yeast is triploid with six chromosomes and the mutants have multiple rearrangements including translocations having the TaqI recognition sequence at the break points. Thus, TAQing2.0 is expected as a useful method to obtain various mutants with altered phenotypes without introducing foreign DNA into asexual industrial microorganisms.


Assuntos
Genoma Fúngico , Transfecção/métodos , Leveduras/genética , DNA Fúngico/genética , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Edição de Genes , Regulação Fúngica da Expressão Gênica , Mutagênese
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