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1.
Front Microbiol ; 14: 1108961, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36846771

RESUMO

Demand for low- or non-alcoholic beers has been growing in recent years. Thus, research is increasingly focusing on non-Saccharomyces species that typically are only able to consume the simple sugars in wort, and therefore have a limited production of alcohol. In this project, new species and strains of non-conventional yeasts were sampled and identified from Finnish forest environments. From this wild yeast collection, a number of Mrakia gelida strains were selected for small-scale fermentation tests and compared with a reference strain, the low-alcohol brewing yeast Saccharomycodes ludwigii. All the M. gelida strains were able to produce beer with an average of 0.7% alcohol, similar to the control strain. One M. gelida strain showing the most promising combination of good fermentation profile and production of desirable flavor active compounds was selected for pilot-scale (40 L) fermentation. The beers produced were matured, filtered, carbonated, and bottled. The bottled beers were then directed for in-house evaluation, and further analyzed for sensory profiles. The beers produced contained 0.6% Alcohol by volume (ABV). According to the sensory analysis, the beers were comparable to those produced by S. ludwigii, and contained detectable fruit notes (banana and plum). No distinct off-flavors were noted. A comprehensive analysis of M. gelida's resistance to temperature extremes, disinfectant, common preservatives, and antifungal agents would suggest that the strains pose little risk to either process hygiene or occupational safety.

2.
Front Microbiol ; 13: 1011155, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36274745

RESUMO

There is a growing trend for beers with novel flavor profiles, as consumers demand a more diversified product range. Such beers can be produced by using non-Saccharomyces yeasts. The yeast species Saccharomycopsis fibuligera is known to produce exceptionally pleasant plum and berry flavors during brewer's wort fermentation while its mycelia growth is most likely a technological challenge in industrial-scale brewing. To better understand and optimize the physiological properties of this yeast species during the brewing process, maltose and maltotriose uptake activity trials were performed. These revealed the existence of active transmembrane transporters for maltose in addition to the known extracellular amylase system. Furthermore, a single cell isolate of S. fibuligera was cultured, which showed significantly less mycelial growth during propagation and fermentation compared to the mother culture and would therefore be much more suitable for application on an industrial scale due to its better flocculation and clarification properties. Genetic differences between the two cultures could not be detected in a (GTG)5 rep-PCR fingerprint and there was hardly any difference in the fermentation process, sugar utilization and flavor profiles of the beers. Accordingly, the characteristic plum and berry flavor could also be perceived by using the culture from the single cell isolate, which was complemented by a dried fruit flavor. A fermentation temperature of 20°C at an original gravity of 10 °P proved to be optimal for producing a low-alcohol beer at around 0.8% (v/v) by applying the S. fibuligera yeast culture from the single cell isolate.

3.
Front Psychiatry ; 12: 735523, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34744825

RESUMO

Background: Anorexia nervosa (AN) is a serious and life-threatening psychiatric condition. With a paucity of approved treatments, there is a desperate need for novel treatment avenues to be explored. Here, we present (1) an overview of the ways through which Public Patient Involvement (PPI) has informed a trial of psilocybin-assisted therapy for AN and (2) a protocol for a pilot study of psilocybin-assisted therapy in AN currently underway at Imperial College London. The study aims to assess the feasibility, brain mechanisms and preliminary outcomes of treating anorexia nervosa with psilocybin. Methods: (1) PPI: Across two online focus groups, eleven individuals with lived experience of AN were presented with an overview of the protocol. Their feedback not only identified solutions to possible barriers for future participants, but also helped the research team to better understand the concept of "recovery" from the perspective of those with lived experience. (2) Protocol: Twenty female participants [21-65 years old, body mass index (BMI) 15 kg/m2 or above] will receive three oral doses of psilocybin (up to 25 mg) over a 6-week period delivered in a therapeutic environment and enveloped by psychological preparation and integration. We will work with participant support networks (care teams and an identified support person) throughout and there will be an extended remote follow-up period of 12 months. Our two-fold primary outcomes are (1) psychopathology (Eating Disorder Examination) across the 6-month follow-up and (2) readiness and motivation to engage in recovery (Readiness and Motivation Questionnaire) across the 6-week trial period. Neurophysiological outcome measures will be: (1) functional magnetic resonance imaging (fMRI) brain changes from baseline to 6-week endpoint and (2) post-acute changes in electroencephalography (EEG) activity, including an electrophysiological marker of neuronal plasticity. Discussion: The results of this pilot study will not only shed light on the acceptability, brain mechanisms, and impression of the potential efficacy of psilocybin as an adjunct treatment for AN but will be essential in shaping a subsequent Randomised Control Trial (RCT) that would test this treatment against a suitable control condition. Clinical Trial Registration: identifier: NCT04505189.

4.
Front Microbiol ; 12: 645271, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33868204

RESUMO

The successful application of Saccharomyces eubayanus and Saccharomyces paradoxus in brewery fermentations has highlighted the potential of wild Saccharomyes yeasts for brewing, and prompted investigation into the application potential of other members of the genus. Here, we evaluate, for the first time, the brewing potential of Saccharomyces jurei. The newly isolated strain from an ash tree (Fraxinus excelsior) in Upper Bavaria, Germany, close to the river Isar, was used to ferment a 12°P wort at 15°C. Performance was compared directly with that of a reference lager strain (TUM 34/70) and the S. eubayanus type strain. Both wild yeast rapidly depleted simple sugars and thereafter exhibited a lag phase before maltose utilization. This phase lasted for 4 and 10 days for S. eubayanus and S. jurei, respectively. S. eubayanus utilized fully the available maltose but, consistent with previous reports, did not use maltotriose. S. jurei, in contrast, utilized approximately 50% of the maltotriose available, making this the first report of maltotriose utilization in a wild Saccharomyces species. Maltotriose use was directly related to alcohol yield with 5.5, 4.9, and 4.5% ABV produced by Saccharomyces pastorianus, S. jurei, and S. eubayanus. Beers also differed with respect to aroma volatiles, with a high level (0.4 mg/L) of the apple/aniseed aroma ethyl hexanoate in S. jurei beers, while S. eubayanus beers had a high level of phenylethanol (100 mg/L). A trained panel rated all beers as being of high quality, but noted clear differences. A phenolic spice/clove note was prominent in S. jurei beer. This was less pronounced in the S. eubayanus beers, despite analytical levels of 4-vinylguaiacol being similar. Tropical fruit notes were pronounced in S. jurei beers, possibly resulting from the high level of ethyl hexanoate. Herein, we present results from the first intentional application of S. jurei as a yeast for beer fermentation (at the time of submission) and compare its fermentation performance to other species of the genus. Results indicate considerable potential for S. jurei application in brewing, with clear advantages compared to other wild Saccharomyces species.

5.
Microorganisms ; 9(3)2021 Mar 02.
Artigo em Inglês | MEDLINE | ID: mdl-33801403

RESUMO

The search for novel brewing strains from non-brewing environments represents an emerging trend to increase genetic and phenotypic diversities in brewing yeast culture collections. Another valuable tool is hybridization, where beneficial traits of individual strains are combined in a single organism. This has been used successfully to create de novo hybrids from parental brewing strains by mimicking natural Saccharomycescerevisiae ale × Saccharomyceseubayanus lager yeast hybrids. Here, we integrated both these approaches to create synthetic hybrids for lager fermentation using parental strains from niches other than beer. Using a phenotype-centered strategy, S. cerevisiae sourdough strains and the S. eubayanus × Saccharomyces uvarum strain NBRC1948 (also referred to as Saccharomyces bayanus) were chosen for their brewing aptitudes. We demonstrated that, in contrast to S. cerevisiae × S. uvarum crosses, hybridization yield was positively affected by time of exposure to starvation, but not by staggered mating. In laboratory-scale fermentation trials at 20 °C, one triple S. cerevisiae × S. eubayanus × S. uvarum hybrid showed a heterotic phenotype compared with the parents. In 2 L wort fermentation trials at 12 °C, this hybrid inherited the ability to consume efficiently maltotriose from NBRC1948 and, like the sourdough S. cerevisiae parent, produced appreciable levels of the positive aroma compounds 3-methylbutyl acetate (banana/pear), ethyl acetate (general fruit aroma) and ethyl hexanoate (green apple, aniseed, and cherry aroma). Based on these evidences, the phenotype-centered approach appears promising for designing de novo lager beer hybrids and may help to diversify aroma profiles in lager beer.

6.
Front Fungal Biol ; 2: 733655, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-37744092

RESUMO

Yeasts in the lager brewing group are closely related and consequently do not exhibit significant genetic variability. Here, an artificial Saccharomyces cerevisiae × Saccharomyces eubayanus tetraploid interspecies hybrid was created by rare mating, and its ability to sporulate and produce viable gametes was exploited to generate phenotypic diversity. Four spore clones obtained from a single ascus were isolated, and their brewing-relevant phenotypes were assessed. These F1 spore clones were found to differ with respect to fermentation performance under lager brewing conditions (15°C, 15 °Plato), production of volatile aroma compounds, flocculation potential and temperature tolerance. One spore clone, selected for its rapid fermentation and acetate ester production was sporulated to produce an F2 generation, again comprised of four spore clones from a single ascus. Again, phenotypic diversity was introduced. In two of these F2 clones, the fermentation performance was maintained and acetate ester production was improved relative to the F1 parent and the original hybrid strain. Strains also performed well in comparison to a commercial lager yeast strain. Spore clones varied in ploidy and chromosome copy numbers, and faster wort fermentation was observed in strains with a higher ploidy. An F2 spore clone was also subjected to 10 consecutive wort fermentations, and single cells were isolated from the resulting yeast slurry. These isolates also exhibited variable fermentation performance and chromosome copy numbers, highlighting the instability of polyploid interspecific hybrids. These results demonstrate the value of this natural approach to increase the phenotypic diversity of lager brewing yeast strains.

7.
Food Microbiol ; 94: 103629, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33279061

RESUMO

De novo sourdough cultures were here assessed for their potential as sources of yeast strains for low-alcohol beer brewing. NGS analysis revealed an abundance of ascomycete yeasts, with some influence of grain type on fungal community composition. Ten different ascomycete yeast species were isolated from different sourdough types (including wheat, rye, and barley) and seven of these were screened for a number of brewing-relevant phenotypes. All seven were maltose-negative and produced less than 1% (v/v) alcohol from a 12 °Plato wort in initial fermentation trials. Strains were further screened for their bioflavouring potential (production of volatile aromas and phenolic notes, reduction of wort aldehydes), stress tolerance (temperature extremes, osmotic stress and ethanol tolerance) and flocculence. Based on these criteria, two species (Kazachstania servazzii and Pichia fermentans) were selected for 10 L-scale fermentation trials and sensory analysis of beers. The latter species was considered particularly suitable for production of low-alcohol wheat beers due to its production of the spice/clove aroma 4-vinylguaiacol, while the former showed potential for lager-style beers due to its clean flavour profile and tolerance to low temperature conditions.


Assuntos
Álcoois/análise , Cerveja/microbiologia , Pão/microbiologia , Maltose/metabolismo , Pichia/metabolismo , Saccharomycetales/metabolismo , Álcoois/metabolismo , Cerveja/análise , Fermentação , Aromatizantes/análise , Aromatizantes/metabolismo , Hordeum/metabolismo , Hordeum/microbiologia , Odorantes , Secale/metabolismo , Secale/microbiologia , Triticum/metabolismo , Triticum/microbiologia
8.
Food Microbiol ; 94: 103640, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33279066

RESUMO

Despite Saccharomyces cerevisiae being a synonym for baker's yeast, the species does not perform well in all baking-related conditions. In particular, dough fermentation, or proofing, is compromised by the species' sensitivity to the low and freezing temperatures that are often used in modern bakeries. Here, screening trials that included representatives of all known Saccharomyces species, showed that S. cerevisiae was generally the most sensitive member of the genus with respect to cold and freezing conditions. We hypothesized therefore that the superior cold tolerance of the non-S. cerevisiae yeast would enable their use as frozen-dough baking strains. To test this, the different yeast species were incorporated into doughs, flash frozen and kept in a frozen state for 14 days. During the proofing stage, dough development was lower in doughs that had been frozen, relative to fresh doughs. This reduction in fermentation performance was however most pronounced with S. cerevisiae. The psychrotolerant yeasts S. eubayanus, S. jurei and S. arboricola showed a strong capacity for post-freeze proofing in terms of dough development and duration of lag phase prior to fermentation. The superior proofing power of these species resulted in breads that were significantly softer and less dense than those prepared with S. cerevisiae. A sensory panel could distinguish the S. cerevisiae and non-S. cerevisiae breads based on their physical properties, but aroma and taste were unaffected by the species employed. To further improve frozen dough baking properties, S. eubayanus, S. jurei and S. arboricola were crossed with baker's yeast through rare mating, and hybrids with improved proofing capacities in both fresh and frozen doughs relative to the parents were created. The use of S. jurei and S. arboricola in baking represents the first potential technological application of these species.


Assuntos
Saccharomyces/química , Saccharomyces/metabolismo , Pão/microbiologia , Fermentação , Microbiologia de Alimentos , Congelamento , Saccharomyces/crescimento & desenvolvimento , Saccharomyces cerevisiae/química , Saccharomyces cerevisiae/crescimento & desenvolvimento , Saccharomyces cerevisiae/metabolismo , Triticum/metabolismo , Triticum/microbiologia
9.
Bioengineering (Basel) ; 7(4)2020 Nov 08.
Artigo em Inglês | MEDLINE | ID: mdl-33171637

RESUMO

Frequently, the oral cavity area can be affected by different diseases, so the patient needs to be submitted to surgery to remove a specific region of the mandibular. A complete or partial discontinuity of the mandibular bone can cause direct or indirect forces variations during the mastication. The dental prosthesis is an alternative to generate an aesthetic or functional solution for oral cavity lesions. However, they can be wrongly designed, or they can lose the adjustment during their useful life, deteriorating the patient's condition. In this work, the influence of the fixation components position for a dental prosthesis will be studied based on the finite element method. By means, it is possible to determine the area of the highest stress concentration generated on the mandibular structure. The temporomandibular image obtained by computational tomography was used as a 3D graphic whole model because in the medical area the morphological factors are extremely important. Vertical loads of 50, 100, 150 and 200 N were applied in three different regions: in the whole buccal cavity, simultaneously in the left and right laterals and only in the right lateral, to determine the values of von Mises stress in the mandible. These results were compared between three finite element software packages (Ansys®, SolidWorks® and Inventor®) and a meshless software (SimSolid®). They showed similar behaviors in the highest mechanical stress concentration in the same regions. Regarding the stress values, the percentage error between each software package was less than 10%. The use of SimSolid® software (meshless) proved to be better at identifying the higher stress generated by the dental prosthesis in the facial skeleton, so its computational efficiency, due to its geometric complexity, was highlighted.

10.
Appl Microbiol Biotechnol ; 103(18): 7597-7615, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31346683

RESUMO

Diastatic strains of Saccharomyces cerevisiae are common contaminants in beer fermentations and are capable of producing an extracellular STA1-encoded glucoamylase. Recent studies have revealed variable diastatic ability in strains tested positive for STA1, and here, we elucidate genetic determinants behind this variation. We show that poorly diastatic strains have a 1162-bp deletion in the promoter of STA1. With CRISPR/Cas9-aided reverse engineering, we show that this deletion greatly decreases the ability to grow in beer and consume dextrin, and the expression of STA1. New PCR primers were designed for differentiation of highly and poorly diastatic strains based on the presence of the deletion in the STA1 promoter. In addition, using publically available whole genome sequence data, we show that the STA1 gene is prevalent among the 'Beer 2'/'Mosaic Beer' brewing strains. These strains utilize maltotriose efficiently, but the mechanisms for this have been unknown. By deleting STA1 from a number of highly diastatic strains, we show here that extracellular hydrolysis of maltotriose through STA1 appears to be the dominant mechanism enabling maltotriose use during wort fermentation in STA1+ strains. The formation and retention of STA1 seems to be an alternative evolutionary strategy for efficient utilization of sugars present in brewer's wort. The results of this study allow for the improved reliability of molecular detection methods for diastatic contaminants in beer and can be exploited for strain development where maltotriose use is desired.


Assuntos
Proteínas Fúngicas/genética , Regiões Promotoras Genéticas , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Amido/metabolismo , Trissacarídeos/metabolismo , Cerveja/microbiologia , Sistemas CRISPR-Cas , Dextrinas/metabolismo , Fermentação , Reprodutibilidade dos Testes , Genética Reversa , Proteínas de Saccharomyces cerevisiae/genética , Deleção de Sequência
11.
Front Microbiol ; 8: 1690, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28943864

RESUMO

The lager brewing yeast Saccharomyces pastorianus, an interspecies hybrid of S. eubayanus and S. cerevisiae, ferments maltotriose, maltose, sucrose, glucose and fructose in wort to ethanol and carbon dioxide. Complete and timely conversion ("attenuation") of maltotriose by industrial S. pastorianus strains is a key requirement for process intensification. This study explores a new evolutionary engineering strategy for improving maltotriose fermentation kinetics. Prolonged carbon-limited, anaerobic chemostat cultivation of the reference strain S. pastorianus CBS1483 on a maltotriose-enriched sugar mixture was used to select for spontaneous mutants with improved affinity for maltotriose. Evolved populations exhibited an up to 5-fold lower residual maltotriose concentration and a higher ethanol concentration than the parental strain. Uptake studies with 14C-labeled sugars revealed an up to 4.75-fold higher transport capacity for maltotriose in evolved strains. In laboratory batch cultures on wort, evolved strains showed improved attenuation and higher ethanol concentrations. These improvements were also observed in pilot fermentations at 1,000-L scale with high-gravity wort. Although the evolved strain exhibited multiple chromosomal copy number changes, analysis of beer made from pilot fermentations showed no negative effects on flavor compound profiles. These results demonstrate the potential of evolutionary engineering for strain improvement of hybrid, alloploid brewing strains.

12.
FEMS Yeast Res ; 17(5)2017 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-28810703

RESUMO

Yeast cryotolerance brings some advantages for wine fermentations, including the improved aromatic complexity of white wines. Naturally cold-tolerant strains are generally less adept at wine fermentation but fermentative fitness can potentially be improved through hybridization. Here we studied the potential of using hybrids involving Saccharomyces eubayanus and a S. cerevisiae wine strain for low-temperature winemaking. Through screening the performance in response to variable concentrations of sugar, nitrogen and temperature, we isolated one hybrid strain that exhibited the superior performance. This hybrid strain was propagated and dried in pilot scale and tested for the fermentation of Macabeu and Sauvignon blanc grape musts. We obtained highly viable active dry yeast, which was able to efficiently ferment the grape musts with superior production of aroma active volatiles, in particular, 2-phenylethanol. The genome sequences of the hybrid strains revealed variable chromosome inheritance among hybrids, particularly within the S. cerevisiae subgenome. With the present paper, we expand the knowledge on the potentialities of using S. eubayanus hybrids in industrial fermentation at beverages other than lager beer.


Assuntos
Quimera/genética , Quimera/metabolismo , Redes e Vias Metabólicas/genética , Saccharomyces/genética , Saccharomyces/metabolismo , Vinho/microbiologia , Metabolismo dos Carboidratos , Temperatura Baixa , Dessecação , Fermentação , Viabilidade Microbiana , Nitrogênio/metabolismo , Saccharomyces/efeitos da radiação , Compostos Orgânicos Voláteis/metabolismo
13.
J Ind Microbiol Biotechnol ; 44(8): 1203-1213, 2017 08.
Artigo em Inglês | MEDLINE | ID: mdl-28451838

RESUMO

Yeast cryotolerance may be advantageous for cider making, where low temperatures are usually employed. Here, we crossed the cryotolerant S. eubayanus with a S. cerevisiae wine strain and assessed the suitability of the hybrids for low-temperature cider fermentation. All strains fermented the juice to 5% ABV, but at different rates; hybrid strains outperformed S. cerevisiae, which was sensitive to low temperatures. The best hybrid fermented similarly to S. eubayanus. S. eubayanus produced sulphurous off flavours which masked a high concentration of fruity ester notes. This phenotype was absent in the hybrid strains, resulting in distinctly fruitier ciders. Aroma was assessed by an independent consumer panel, which rated the hybrid ciders as identical to the wine strain cider. Both were significantly more pleasant than the S. eubayanus cider. Interspecific hybridization can apparently be used effectively to improve low-temperature fermentation performance without compromising product quality.


Assuntos
Bebidas Alcoólicas/microbiologia , Fermentação , Microbiologia de Alimentos , Saccharomyces cerevisiae/metabolismo , Saccharomyces/metabolismo , Adolescente , Adulto , Idoso , Temperatura Baixa , Comportamento do Consumidor , Feminino , Manipulação de Alimentos , Qualidade dos Alimentos , Sucos de Frutas e Vegetais/microbiologia , Humanos , Hibridização Genética , Masculino , Pessoa de Meia-Idade , Mitocôndrias/metabolismo , Odorantes/análise , Saccharomyces/classificação , Saccharomyces/genética , Saccharomyces cerevisiae/genética , Paladar , Compostos Orgânicos Voláteis/análise , Adulto Jovem
14.
Appl Microbiol Biotechnol ; 101(1): 65-78, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27885413

RESUMO

The natural interspecies Saccharomyces cerevisiae × Saccharomyces eubayanus hybrid yeast is responsible for global lager beer production and is one of the most important industrial microorganisms. Its success in the lager brewing environment is due to a combination of traits not commonly found in pure yeast species, principally low-temperature tolerance, and maltotriose utilization. Parental transgression is typical of hybrid organisms and has been exploited previously for, e.g., the production of wine yeast with beneficial properties. The parental strain S. eubayanus has only been discovered recently and newly created lager yeast strains have not yet been applied industrially. A number of reports attest to the feasibility of this approach and artificially created hybrids are likely to have a significant impact on the future of lager brewing. De novo S. cerevisiae × S. eubayanus hybrids outperform their parent strains in a number of respects, including, but not restricted to, fermentation rate, sugar utilization, stress tolerance, and aroma formation. Hybrid genome function and stability, as well as different techniques for generating hybrids and their relative merits are discussed. Hybridization not only offers the possibility of generating novel non-GM brewing yeast strains with unique properties, but is expected to aid in unraveling the complex evolutionary history of industrial lager yeast.


Assuntos
Bebidas Alcoólicas/microbiologia , Cruzamentos Genéticos , Saccharomyces/genética , Saccharomyces/metabolismo , Fermentação , Microbiologia Industrial , Engenharia Metabólica
15.
FEMS Yeast Res ; 16(5)2016 08.
Artigo em Inglês | MEDLINE | ID: mdl-27364826

RESUMO

Brewer's wort is a challenging environment for yeast as it contains predominantly α-glucoside sugars. There exist two subgroups of the lager yeast Saccharomyces pastorianus which differ in sugar utilisation. We performed wort fermentations and compared representative strains from both groups with respect to their ability to transport and ferment maltose and maltotriose. Additionally, we mapped the transporters MALx1, AGT1, MPHx and MTT1 by Southern blotting. Contrary to previous observations, group I comprises a diverse set of strains, with varying ability to transport and ferment maltotriose. Of the eight group I strains, three efficiently utilised maltotriose, a property enabled by the presence of transmembrane transporters SeAGT1 and MTT1 A58, a variant of the group I type strain (CBS1513) performed particularly well, taking up maltotriose at a higher rate than maltose and retaining significant transport activity at temperatures as low as 0°C. Analysis of transporter distribution in this strain revealed an increased copy number of the MTT1 gene, which encodes the only permease known with higher affinity for maltotriose than maltose and low temperature dependence for transport. We propose that much of the variation in lager yeast fermentation behaviour is determined by the presence or absence of specific transmembrane transporters.


Assuntos
Maltose/metabolismo , Saccharomyces/metabolismo , Trissacarídeos/metabolismo , Southern Blotting , Mapeamento Cromossômico , Fermentação , Dosagem de Genes , Proteínas de Membrana Transportadoras/genética , Saccharomyces/genética , Temperatura
16.
Appl Microbiol Biotechnol ; 100(16): 7203-22, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27183995

RESUMO

The genomes of hybrid organisms, such as lager yeast (Saccharomyces cerevisiae × Saccharomyces eubayanus), contain orthologous genes, the functionality and effect of which may differ depending on their origin and copy number. How the parental subgenomes in lager yeast contribute to important phenotypic traits such as fermentation performance, aroma production, and stress tolerance remains poorly understood. Here, three de novo lager yeast hybrids with different ploidy levels (allodiploid, allotriploid, and allotetraploid) were generated through hybridization techniques without genetic modification. The hybrids were characterized in fermentations of both high gravity wort (15 °P) and very high gravity wort (25 °P), which were monitored for aroma compound and sugar concentrations. The hybrid strains with higher DNA content performed better during fermentation and produced higher concentrations of flavor-active esters in both worts. The hybrid strains also outperformed both the parent strains. Genome sequencing revealed that several genes related to the formation of flavor-active esters (ATF1, ATF2¸ EHT1, EEB1, and BAT1) were present in higher copy numbers in the higher ploidy hybrid strains. A direct relationship between gene copy number and transcript level was also observed. The measured ester concentrations and transcript levels also suggest that the functionality of the S. cerevisiae- and S. eubayanus-derived gene products differs. The results contribute to our understanding of the complex molecular mechanisms that determine phenotypes in lager yeast hybrids and are expected to facilitate targeted strain development through interspecific hybridization.


Assuntos
Cerveja/microbiologia , Quimera/genética , Etanol/metabolismo , Fermentação/genética , Saccharomyces cerevisiae/genética , Quimera/crescimento & desenvolvimento , DNA Fúngico/genética , Ésteres/análise , Hibridização Genética , Compostos Orgânicos/análise , Ploidias , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Saccharomyces cerevisiae/classificação , Saccharomyces cerevisiae/metabolismo , Transcrição Gênica/genética
17.
Acta Med Port ; 29(9): 507-513, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28060687

RESUMO

INTRODUCTION: Though typically found in adolescents, acne frequently affects the adult population. In Portugal, no representative studies about acne prevalence in greater populational areas are known. This study had the following objectives: calculate acne prevalence in primary healthcare patients; its characteristics, including risk factors for onset and worsening; collect information concerning selfmedication and/or prescribed treatment. MATERIAL AND METHODS: Application of a questionnaire and physical examination into a convenience sample, composed by adults from20 - 60 years old that visited, in random days, five primary healthcare centers of the Greater Porto area. RESULTS: From the 1,055 evaluated adults, acne prevalence was estimated at 61.5%. Only 36.8% of acne suffering adults was awareof their condition. More than half of tobacco users (62.3%) had acne. Just 25% of acne sufferers sought treatment. The most affected areas were the malars, differing from the younger disease form. DISCUSSION: Acne prevalence was relatively higher than other studies. The results of this study have confirmed that acne seems to be related with increased calorie intake, tobacco smoking, whole milk and fat-reduced milk consumption, hirsutism, alopecia and seborrhea. CONCLUSION: In the primary healthcare set, more than half of adults were acne sufferers. These numbers show how important primary healthcare services can be in preventing and intervening when managing acne-related risk factors and psychosocial consequences.


Introdução: A acne, apesar de caraterística no adolescente, afeta frequentemente o adulto. Em Portugal, não são conhecidos estudos sobre a prevalência da acne no adulto, nomeadamente observada nos grandes centros populacionais. Os objetivos deste estudo foram estimar a prevalência da acne nos utentes dos serviços de cuidados de saúde primários, as suas características, incluindo fatores de risco para início e/ou agravamento, além de coletar informação relacionada com a automedicação e/ou tratamento prescrito. Material e Métodos: Aplicação de questionário e exame físico a amostra composta por adultos entre os 20 - 60 anos de idade que visitaram, em dias aleatórios, cinco centros de saúde da área do Grande Porto. Resultados: Dos 1 055 adultos avaliados, a prevalência estimada de acne foi de 61,5%. Apenas 36,8% dos portadores estavam cientes da sua condição e somente 25% afirmou ter procurado tratamento. Mais de metade dos fumadores (62,3%) apresentavam acne. A zona mais afetada foi a região malar, diferindo da forma adolescente da doença. Discussão: A prevalência encontrada de acne foi mais elevada do que em outros estudos. A acne aparentou estar relacionada com a ingestão calórica aumentada, tabagismo, consumo de leite gordo ou meio gordo, hirsutismo, alopécia e seborreia. Conclusão: No contexto dos cuidados de saúde primários, mais de metade dos adultos eram portadores de acne. Estes números demonstram o quão importante pode ser a prevenção e intervenção pelos profissionais dos cuidados de saúde primários, tanto para a redução de eventuais factores de risco como para a intervenção terapêutica e consequências psicossociais relacionadas com a acne. Palavras-chave: Acne Vulgaris/epidemiologia; Adulto; Cuidados de Saúde Primários; Portugal; Prevalência.


Assuntos
Acne Vulgar/epidemiologia , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Portugal/epidemiologia , Prevalência , Atenção Primária à Saúde , Fatores de Risco , Adulto Jovem
18.
Yeast ; 32(10): 615-28, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26129747

RESUMO

Genes encoding L-arabinose transporters in Kluyveromyces marxianus and Pichia guilliermondii were identified by functional complementation of Saccharomyces cerevisiae whose growth on L-arabinose was dependent on a functioning L-arabinose transporter, or by screening a differential display library, respectively. These transporters also transport D-xylose and were designated KmAXT1 (arabinose-xylose transporter) and PgAXT1, respectively. Transport assays using L-arabinose showed that KmAxt1p has K(m) 263 mM and V(max) 57 nM/mg/min, and PgAxt1p has K(m) 0.13 mM and V(max) 18 nM/mg/min. Glucose, galactose and xylose significantly inhibit L-arabinose transport by both transporters. Transport assays using D-xylose showed that KmAxt1p has K(m) 27 mM and V(max) 3.8 nM/mg/min, and PgAxt1p has K(m) 65 mM and V(max) 8.7 nM/mg/min. Neither transporter is capable of recovering growth on glucose or galactose in a S. cerevisiae strain deleted for hexose and galactose transporters. Transport kinetics of S. cerevisiae Gal2p showed K(m) 371 mM and V(max) 341 nM/mg/min for L-arabinose, and K(m) 25 mM and V(max) 76 nM/mg/min for galactose. Due to the ability of Gal2p and these two newly characterized transporters to transport both L-arabinose and D-xylose, one scenario for the complete usage of biomass-derived pentose sugars would require only the low-affinity, high-throughput transporter Gal2p and one additional high-affinity general pentose transporter, rather than dedicated D-xylose or L-arabinose transporters. Additionally, alignment of these transporters with other characterized pentose transporters provides potential targets for substrate recognition engineering.


Assuntos
Arabinose/metabolismo , Proteínas Fúngicas/genética , Kluyveromyces/genética , Proteínas de Membrana Transportadoras/genética , Pichia/genética , Saccharomyces cerevisiae/crescimento & desenvolvimento , Xilose/metabolismo , Arabinose/química , Transporte Biológico , Proteínas Fúngicas/química , Proteínas Fúngicas/metabolismo , Expressão Gênica , Engenharia Genética , Cinética , Proteínas de Membrana Transportadoras/química , Proteínas de Membrana Transportadoras/metabolismo , Saccharomyces cerevisiae/química , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Xilose/química
20.
J Ind Microbiol Biotechnol ; 42(5): 769-78, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25682107

RESUMO

The interspecific hybrid Saccharomyces pastorianus is the most commonly used yeast in brewery fermentations worldwide. Here, we generated de novo lager yeast hybrids by mating a domesticated and strongly flocculent Saccharomyces cerevisiae ale strain with the Saccharomyces eubayanus type strain. The hybrids were characterized with respect to the parent strains in a wort fermentation performed at temperatures typical for lager brewing (12 °C). The resulting beers were analysed for sugar and aroma compounds, while the yeasts were tested for their flocculation ability and α-glucoside transport capability. These hybrids inherited beneficial properties from both parent strains (cryotolerance, maltotriose utilization and strong flocculation) and showed apparent hybrid vigour, fermenting faster and producing beer with higher alcohol content (5.6 vs 4.5 % ABV) than the parents. Results suggest that interspecific hybridization is suitable for production of novel non-GM lager yeast strains with unique properties and will help in elucidating the evolutionary history of industrial lager yeast.


Assuntos
Cerveja/microbiologia , Hibridização Genética , Saccharomyces/genética , Cerveja/análise , Evolução Biológica , Transporte Biológico , Etanol/análise , Fermentação , Floculação , Indústria Alimentícia , Glucosídeos/metabolismo , Vigor Híbrido/genética , Saccharomyces/metabolismo , Especificidade da Espécie , Trissacarídeos/metabolismo
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