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1.
J Agric Food Chem ; 67(37): 10423-10431, 2019 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-31487168

RESUMO

Plants often produce antifungal peptides and proteins in response to infection. Also wheat, which is the main ingredient of bread dough, contains such components. Here, we show that while some industrial strains of the baker's yeast Saccharomyces cerevisiae can efficiently ferment dough, some other strains show much lower fermentation capacities because they are sensitive to a specific wheat protein. We purified and identified what turned out to be a thaumatin-like protein through a combination of activity-guided fractionation, cation exchange chromatography, reversed-phase HPLC, and LC-MS/MS. Recombinant expression of the corresponding gene and testing the activity confirmed the inhibitory activity of the protein.


Assuntos
Proteínas de Plantas/química , Proteínas de Plantas/farmacologia , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/crescimento & desenvolvimento , Triticum/química , Cromatografia Líquida , Fermentação , Proteínas de Plantas/isolamento & purificação , Proteínas de Plantas/metabolismo , Saccharomyces cerevisiae/metabolismo , Espectrometria de Massas em Tandem , Triticum/genética , Triticum/metabolismo , Triticum/microbiologia
2.
Microb Cell Fact ; 18(1): 160, 2019 Sep 23.
Artigo em Inglês | MEDLINE | ID: mdl-31547812

RESUMO

BACKGROUND: Alpha-Terpineol (α-Terpineol), a C10 monoterpenoid alcohol, is widely used in the cosmetic and pharmaceutical industries. Construction Saccharomyces cerevisiae cell factories for producing monoterpenes offers a promising means to substitute chemical synthesis or phytoextraction. RESULTS: α-Terpineol was produced by expressing the truncated α-Terpineol synthase (tVvTS) from Vitis vinifera in S. cerevisiae. The α-Terpineol titer was increased to 0.83 mg/L with overexpression of the rate-limiting genes tHMG1, IDI1 and ERG20F96W-N127W. A GSGSGSGSGS linker was applied to fuse ERG20F96W-N127W with tVvTS, and expressing the fusion protein increased the α-Terpineol production by 2.87-fold to 2.39 mg/L when compared with the parental strain. In addition, we found that farnesyl diphosphate (FPP) accumulation by down-regulation of ERG9 expression and deletion of LPP1 and DPP1 did not improve α-Terpineol production. Therefore, ERG9 was overexpressed and the α-Terpineol titer was further increased to 3.32 mg/L. The best α-Terpineol producing strain LCB08 was then used for batch and fed-batch fermentation in a 5 L bioreactor, and the production of α-Terpineol was ultimately improved to 21.88 mg/L. CONCLUSIONS: An efficient α-Terpineol production cell factory was constructed by engineering the S. cerevisiae mevalonate pathway, and the metabolic engineering strategies could also be applied to produce other valuable monoterpene compounds in yeast.


Assuntos
Cicloexenos/metabolismo , Engenharia Metabólica , Monoterpenos/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Fosfatos de Poli-Isoprenil/metabolismo , Sesquiterpenos/metabolismo , Vitis/enzimologia , Vitis/genética
3.
Shokuhin Eiseigaku Zasshi ; 60(4): 88-95, 2019.
Artigo em Japonês | MEDLINE | ID: mdl-31474656

RESUMO

Microbial colony counts of concern of food products are one of the most important items in microbiological examinations. The distributions of colony counts per agar plate of food samples are considered to be reflected with microbial cell distributions in food homogenates. However, (i) the probabilistic distributions of the colony counts per agar plate at the dilution of counting and (ii) the relationship between the colony counts per plate and the number of agar plates for food samples have not been intensively studied so far. In this study, therefore, these two points were studied with raw food samples of raw minced beef and chicken and raw milk and microbial culture samples of Escherichia coli, Staphylococcus aureus, and Saccharomyces cerevisiae. Among four major probabilistic distributions, it was found that aerobic plate counts per plate of the foods were well described with negative binomial, Poisson, and normal distributions and that the colony counts per plate of microbial cultures were described well with binomial, Poisson, and normal distributions. The effect of the number of agar plates on the estimation of the mean of colony counts per plate of a sample was then studied with the data randomly resampled from the experimental data. The resampled data showed that with more number of plates the mean of counts fluctuated less and the coefficients of variation of colony counts per plate decreased further, which were coincident to the estimated by the central limit theory. Our study would provide useful information on the characteristics of colony counts per plate of food samples which are routinely examined.


Assuntos
Contagem de Colônia Microbiana , Contaminação de Alimentos/análise , Microbiologia de Alimentos , Ágar , Animais , Bovinos , Escherichia coli/isolamento & purificação , Carne/microbiologia , Leite/microbiologia , Saccharomyces cerevisiae/isolamento & purificação , Staphylococcus aureus/isolamento & purificação
4.
Bioresour Technol ; 291: 121844, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31400704

RESUMO

A low-temperature sodium hydroxide (NaOH) pretreatment for sugarcane bagasse (SCB) was obtained via the surface response design in this study. However, a large quantity of water consumption and wastewater generation which have been the common problems for alkaline pretreatment of lignocellulose still exists in this pretreatment. In order to reduce water consumption and wastewater generation, this study attempted to perform enzymatic hydrolysis and fermentation of NaOH-treated SCB without washing process. It showed that after pretreatment and solid-liquid separation, NaOH-treated SCB could be directly hydrolysed by cellulase via pH and solid-liquid adjustment without washing steps, and the maximum enzymatic hydrolysis efficiency could reach to 70.2%. A domesticated Saccharomyces cerevisiae Y2034 which can endure 6-times diluted BL was obtained, and realized 67.5% ethanol yield from the enzymatic hydrolysate of unwashed NaOH-treated SCB. It provided a clue for converting NaOH-treated lignocellulose to ethanol at low water consumption and wastewater generation.


Assuntos
Celulose/química , Etanol/química , Saccharum/química , Hidróxido de Sódio/química , Celulose/metabolismo , Temperatura Baixa , Etanol/metabolismo , Fermentação , Hidrólise , Lignina/química , Lignina/metabolismo , Saccharomyces cerevisiae/metabolismo , Saccharum/metabolismo , Águas Residuárias/química
5.
Gene ; 717: 144046, 2019 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-31434006

RESUMO

Flavonoids are major polyphenol compounds in plant secondary metabolism. The hydroxylation pattern of the B-ring of flavonoids is determined by the flavonoid 3'-hydroxylase (F3'H) and flavonoid 3',5'-hydroxylase (F3'5'H). In this paper, one CsF3'H and two CsF3'5'Hs (CsF3'5'Ha and CsF3'5'Hb) were isolated. The phylogenetic tree results showed that F3'H and F3'5'Hs belong to the CYP75B and CYP75A, respectively. The Expression pattern analysis showed that the expression of CsF3'5'Ha and CsF3'5'Hb in the bud and 1st leaf were higher than other tissues. However, the CsF3'H had the highest expression in the 4th and mature leaf. The correlation analysis showed that the expression of CsF3'5'Hs is positively associated with the concentration of B-trihydroxylated catechins, and the expression of CsF3'H is positively associated with the Q contentration. Heterologous expression of these genes in yeast showed that CsF3'H and CsF3'5'Ha can catalyze flavanones, flavonols and flavanonols to the corresponding 3', 4' or 3', 4', 5'-hydroxylated compounds, for which the optimum substrate is naringenin. The enzyme of CsF3'5'Hb can only catalyze flavonols (including K and Q) and flavanonols (DHK and DHQ), of which the highest activities in catalyzing are DHK. Interestingly, The experiment of site-directed mutagenesis suggested that two novel sites near the C-terminal were discovered impacting on the activity of the CsF3'5'H. These results provide a significantly molecular basis on the accumulation B-ring hydroxylation of flavonoids in tea plant.


Assuntos
Camellia sinensis/genética , Sistema Enzimático do Citocromo P-450/genética , Flavonoides/metabolismo , Camellia sinensis/metabolismo , Clonagem Molecular , Sistema Enzimático do Citocromo P-450/metabolismo , Flavonoides/química , Regulação da Expressão Gênica de Plantas , Hidroxilação , Mutagênese Sítio-Dirigida , Filogenia , Folhas de Planta/genética , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Saccharomyces cerevisiae/genética
6.
World J Microbiol Biotechnol ; 35(9): 136, 2019 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-31432249

RESUMO

Volatile phenols such as 4-ethylphenol are produced from hydroxycinnamic acids by Dekkera bruxellensis, an important yeast contaminating alcoholic fermentations. 4-ethylphenol results from the decarboxylation and reduction of p-coumaric acid, a compound found in sugarcane musts. In wine, volatile phenols are responsible by sensorial alterations whereas in the context of bioethanol fermentation, little is known about their effects on the main yeast, Saccharomyces cerevisiae. Here we evaluated the interaction of 4-ethylphenol and pH, sucrose and ethanol on the growth and fermentation capacity of the industrial strain of S. cerevisiae PE-2. A central compound rotational design was utilized to evaluate the effect of 4-ethylphenol, pH, ethanol and sucrose concentration on the yeast maximum specific growth rate (µmax) in microplate experiments in YPS medium (Yeast extract-Peptone-Sucrose), at 30 °C. Following, single-cycle fermentations in YPS medium, pH 4.5, 17% sucrose, at 30 °C, with 4-ethylphenol in concentrations of 10 and 20 mg L-1 being added at the start or after 4 h of fermentation, were carried out. 4-ethylphenol affected µmax of S. cerevisiae in situations that resemble the conditions of industrial bioethanol production, especially the low pH of the fermentation medium and the high ethanol concentration because of the anaerobic sucrose uptake. The addition of 4-ethylphenol on fermentation resulted in significant effect on the cell yeast concentration, pH and alcohol production, with significant decrease from 86% to the range of 65-74% in the fermentative efficiency. The industrial yeast S. cerevisiae PE-2 growth and fermentative capacity were affected by the presence of 4-ethylphenol, a metabolite produced by D. bruxellensis, which may contribute to explain the impact of this yeast on bioethanol industrial production.


Assuntos
Etanol/metabolismo , Fermentação , Microbiologia Industrial , Fenóis/metabolismo , Saccharomyces cerevisiae/crescimento & desenvolvimento , Saccharomyces cerevisiae/metabolismo , Sacarose/metabolismo , Meios de Cultura/química , Inibidores do Crescimento/metabolismo , Concentração de Íons de Hidrogênio , Saccharomyces cerevisiae/efeitos dos fármacos , Temperatura Ambiente
7.
Microbiol Res ; 227: 126298, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31421716

RESUMO

An increasing number of infections originating from probiotic use are reported worldwide, with the majority of such cases caused by the yeast Saccharomyces 'boulardii', a subtype of S. cerevisiae. Reliably linking infectious cases to probiotic products requires unequivocal genotyping data, however, these techniques are often time-consuming and difficult to implement in routine diagnostics. This leads to a widespread lack of genetic data regarding the origin of Saccharomyces infections. We propose a quick and reliable PCR-based protocol for the identification of S. 'boulardii' based on a combined analysis of interdelta fingerprinting and microsatellite typing. By applying various typing methods and our proposed method to the clinical yeast collection of a Hungarian hospital we show that probiotic origin is common among clinical Saccharomyces, and that the new multiplex method enables rapid and unequivocal identification of probiotic yeast infections. This method can be applied for the identification of yeast infection sources, helping decisions on probiotic use.


Assuntos
Reação em Cadeia da Polimerase Multiplex/métodos , Técnicas de Tipagem Micológica/métodos , Probióticos , Saccharomyces/genética , Saccharomyces/isolamento & purificação , DNA Fúngico/isolamento & purificação , Fungemia/microbiologia , Técnicas de Genotipagem , Humanos , Repetições de Microssatélites , Micoses/microbiologia , Saccharomyces/classificação , Saccharomyces/patogenicidade , Saccharomyces cerevisiae/classificação , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/isolamento & purificação
8.
J Agric Food Chem ; 67(37): 10521-10533, 2019 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-31461284

RESUMO

This work was designed to comparatively investigate 27 dietary flavonoids that act as α-glucosidase inhibitors and insulin sensitizers. On the basis of the results of an in vitro experiment of α-glucosidase inhibition, myricetin (IC50 = 11.63 ± 0.36 µM) possessed the strongest inhibitory effect, followed by apigenin-7-O-glucoside (IC50 = 22.80 ± 0.24 µM) and fisetin (IC50 = 46.39 ± 0.34 µM). A three-dimensional quantitative structure-activity relationship model of α-glucosidase inhibitors with good predictive capability [comparative molecular field analysis, q2 = 0.529, optimum number of components (ONC) = 10, R2 = 0.996, F = 250.843, standard error of estimation (SEE) = 0.064, and two descriptors; comparative similarity index analysis, q2 = 0.515, ONC = 10, R2 = 0.997, F = 348.301, SEE = 0.054, and four descriptors] was established and indicated that meta positions of ring B favored bulky and minor, electron-withdrawing, and hydrogen bond donor groups. The presence of electron-donating and hydrogen bond acceptor groups at position 4' of ring B could improve α-glucosidase activity. Position 3 of ring C favored minor, electron-donating, and hydrogen bond donor groups, whereas position 7 of ring A favored bulky and hydrogen bond acceptor groups. Molecular docking screened five flavonoids (baicalein, isorhamnetin-3-O-rutinoside, apigenin-7-O-glucoside, kaempferol-7-O-ß-glucoside, and cyanidin-3-O-glucoside) that can act as insulin sensitizers and form strong combinations with four key protein targets involved in the insulin signaling pathway. Apigenin-7-O-glucoside (60 µM) can effectively improve insulin resistance, and glucose uptake increased by approximately 73.06% relative to the model group of insulin-resistant HepG2 cells. Therefore, apigenin-7-O-glucoside might serve as the most effective α-glucosidase inhibitor and insulin sensitizer. This work may guide diabetes patients to improve their condition through dietary therapy.


Assuntos
Flavonoides/química , Inibidores de Glicosídeo Hidrolases/química , Insulina/metabolismo , Flavonoides/metabolismo , Flavonoides/farmacologia , Inibidores de Glicosídeo Hidrolases/metabolismo , Inibidores de Glicosídeo Hidrolases/farmacologia , Células Hep G2 , Humanos , Simulação de Acoplamento Molecular , Saccharomyces cerevisiae/enzimologia , Proteínas de Saccharomyces cerevisiae/química , Proteínas de Saccharomyces cerevisiae/metabolismo , alfa-Glucosidases/química , alfa-Glucosidases/metabolismo
9.
Bioresour Technol ; 292: 121922, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31398543

RESUMO

The limited tolerance of Saccharomyces cerevisiae to the inhibitors present in lignocellulosic hydrolysates is a major challenge in second-generation bioethanol production. Short-term adaptation of the yeast to lignocellulosic hydrolysates during cell propagation has been shown to improve its tolerance, and thus its performance in lignocellulose fermentation. The aim of this study was to investigate the short-term adaptation effects in yeast strains with different genetic backgrounds. Fed-batch propagation cultures were supplemented with 40% wheat straw hydrolysate during the feed phase to adapt two different pentose-fermenting strains, CR01 and KE6-12. The harvested cells were used to inoculate fermentation media containing 80% or 90% wheat straw hydrolysate. The specific ethanol productivity during fermentation was up to 3.6 times higher for CR01 and 1.6 times higher for KE6-12 following adaptation. The influence of physiological parameters such as viability, storage carbohydrate content, and metabolite yields following short-term adaptation demonstrated that short-term adaptation was strain dependent.


Assuntos
Saccharomyces cerevisiae , Xilose , Etanol , Fermentação , Pentoses
10.
Chem Biol Interact ; 311: 108776, 2019 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-31369745

RESUMO

Omeprazole (OM), a prototype proton pump inhibitor, oxidizes thiol groups and induces DNA damage. The aim of this study was to evaluate the oxidative effects of omeprazole and its interactions with ascorbic acid (AA, 50 µM) and retinol palmitate (RP) in proficient and deficient Saccharomyces cerevisiae strains, as well as levels of cytogenetic damage in Sarcoma 180 (S180) cells. Omeprazole was tested at concentrations of 10, 20 and 40 µg/mL, whereas H2O2 (10 mM), cyclophosphamide (20 mg/mL), and saline (0.9% NaCl solution) were employed as stressor, positive control, and negative control, respectively. Results revealed that omeprazole concentration-dependently induces oxidative effects in S. cerevisiae strains. However, omeprazole co-treated with ascorbic acid (50 µM) and retinol palmitate (100 IU) significantly modulated the oxidative damage inflected on the S. cerevisiae strains. Furthermore, omeprazole did not produce micronucleus formation and chromosomal bridges in S180 cells, but induced shoots. Significant increase in karyolysis and karyorrhexis were also observed with the omeprazole treated groups, which was modulated by co-treatment with ascorbic acid and retinol palmitate. Taken all together, it is suggested that ascorbic acid and retinol palmitate can substantially modulate the oxidative damage caused by omeprazole on the S. cerevisiae strains, however, much precaution is recommended with omeprazole and antioxidant co-treatment.


Assuntos
Ácido Ascórbico/farmacologia , Aberrações Cromossômicas/efeitos dos fármacos , Omeprazol/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Saccharomyces cerevisiae/efeitos dos fármacos , Vitamina A/análogos & derivados , Animais , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Ciclofosfamida/toxicidade , Peróxido de Hidrogênio/toxicidade , Camundongos , Testes para Micronúcleos , Vitamina A/farmacologia
11.
J Environ Manage ; 248: 109295, 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31376612

RESUMO

Microbial contamination of vegetables due to irrigation with wastewater-polluted streams is a common problem around most cities in developing countries because wastewater is an available source of water and nutrients but wastewater treatment is often inadequate. On-farm treatment of polluted water is a feasible option to manage microbial risks in a multi-barrier approach. Current evidence indicates good suitability of biochar filters for microbe removal from wastewater using the hydraulic loading rate (HLR) designed for sand filters, but their suitability has not been tested under on-farm conditions. This study evaluated the combined effect of several variables on removal of microbial indicators from diluted wastewater by biochar filtration on-farm and the correlations between removal efficiency and HLR. Columns of biochar with three different effective particle diameters (d10) were fed with diluted wastewater at 1x, 6x, and 12x the design HLR and two levels of water salinity (electrical conductivity, EC). Influent and effluent samples were collected from the columns and analyzed for bacteriophages (ɸX174 and MS2), Escherichia coli, Enterococcus spp., and Saccharomyces cerevisiae. Microbe removal decreased with increasing HLR, from 2 to 4 to 1 log10 for bacteria and from 2 to 0.8 log10 for viruses, while S. cerevisiae removal was unaffected. Effective particle diameter (d10) was the main variable explaining microbe removal at 6x and 12x, while EC had no effect. Correlation analysis showed removal of 2 log10 bacteria and 1 log10 virus at 3x HLR. Thus biochar filters on-farm would not remove significant amounts of bacteria and viruses. However, the design HLR was found to be conservative. These results, and some technical and management considerations identified, can assist in the development of a scientific method for designing biochar filters for on-farm and conventional wastewater treatment.


Assuntos
Eliminação de Resíduos Líquidos , Águas Residuárias , Carvão Vegetal , Fazendas , Filtração , Saccharomyces cerevisiae
12.
World J Microbiol Biotechnol ; 35(7): 111, 2019 Jul 06.
Artigo em Inglês | MEDLINE | ID: mdl-31280424

RESUMO

Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) immune systems in bacteria have been used as tools for genome engineering. Thus far, the CRISPR-Cas system has been used in various yeast, bacterial, and mammalian cells. Saccharomyces cerevisiae is a nonpathogenic yeast, classified under "generally recognized as safe", and has long been used to produce consumables such as alcohol or bread. Additionally, recombinant cells of S. cerevisiae have been constructed and used to produce various bio-based chemicals. Some types of CRISPR-Cas system for genetic manipulation have been constructed during the early developmental stages of the CRISPR-Cas system and have been mainly used for gene knock-in and knock-out manipulations. Thereafter, these systems have been used for various novel purposes such as metabolic engineering and tolerance engineering. In this review, we have summarized different aspects of the CRISPR-Cas in the yeast S. cerevisiae, from its basic principles to various applications. This review describes the CRISPR system in S. cerevisiae based on the differences in its origin and efficiency followed by its basic applications; for example, its involvement in gene knock-in and knock-out has been outlined. Finally, advanced applications of the CRISPR system in the bioproduction of useful chemicals have been summarized.


Assuntos
Sistemas CRISPR-Cas , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas , Engenharia Metabólica/métodos , Saccharomyces cerevisiae/metabolismo , Edição de Genes/métodos , Regulação Fúngica da Expressão Gênica , Técnicas de Introdução de Genes/métodos , Técnicas de Inativação de Genes/métodos , Saccharomyces cerevisiae/genética
13.
World J Microbiol Biotechnol ; 35(7): 112, 2019 Jul 08.
Artigo em Inglês | MEDLINE | ID: mdl-31286266

RESUMO

Microorganisms have evolved permeases to incorporate various essential nutrients and exclude harmful products, which assists in adaptation to different environmental conditions for survival. As permeases are directly involved in the utilization of and regulatory response to nutrient sources, metabolic engineering of microbial permeases can predictably influence nutrient metabolism and regulation. In this mini-review, we have summarized the mechanisms underlying the general regulation of permeases, and the current advancements and future prospects of metabolic engineering strategies targeting the permeases in Saccharomyces cerevisiae. The different types of permeases and their regulatory mechanisms have been discussed. Furthermore, methods for metabolic engineering of permeases have been highlighted. Understanding the mechanisms via which permeases are meticulously regulated and engineered will not only facilitate research on regulation of global nutrition and yeast metabolic engineering, but can also provide important insights for future studies on the synthesis of valuable products and elimination of harmful substances in S. cerevisiae.


Assuntos
Proteínas de Membrana Transportadoras/metabolismo , Engenharia Metabólica/métodos , Saccharomyces cerevisiae/metabolismo , Transporte Biológico , Carbono/metabolismo , Glucose/metabolismo , Proteínas de Membrana Transportadoras/genética , Nitrogênio/metabolismo , Saccharomyces cerevisiae/genética
14.
J Agric Food Chem ; 67(32): 8986-8993, 2019 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-31347835

RESUMO

Trehalose plays a crucial role in response to freezing stress in baker's yeast. MAL62, a gene involved in the adenosine diphosphoglucose-dependent trehalose synthesis pathway, can increase trehalose content. However, the difference between MAL62-related trehalose synthesis and traditional uridine diphosphoglucose-dependent trehalose synthesis is not well-understood. MAL62 overexpression showed less effect in enhancing intracellular trehalose compared to TPS1 overexpression. However, MAL62 overexpression elicited trehalose synthesis before fermentation with enhanced maltose metabolism and had a similar effect on cell viability after freezing. Furthermore, MAL62 and TPS1 overexpression in the NTH1 deletion background further strengthened freezing tolerance and improved leavening ability. Our results suggest that the enhancement in freezing tolerance by MAL62 overexpression may involve multiple pathways rather than simply enhancing trehalose synthesis. The results reveal valuable insights into the relationship between maltose metabolism and freezing tolerance and may help to develop better yeast strains for enhancing fermentation characteristics of frozen dough.


Assuntos
Glucosiltransferases/metabolismo , Maltose/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/enzimologia , Saccharomyces cerevisiae/genética , alfa-Glucosidases/metabolismo , Farinha/análise , Farinha/microbiologia , Congelamento , Regulação Fúngica da Expressão Gênica , Glucosiltransferases/genética , Saccharomyces cerevisiae/química , Proteínas de Saccharomyces cerevisiae/genética , Trealase/genética , Trealase/metabolismo , Trealose/metabolismo , alfa-Glucosidases/genética
15.
Bioengineered ; 10(1): 335-344, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31322471

RESUMO

Selenium-enriched yeast can transform toxic inorganic selenium into absorbable organic selenium, which is of great significance for human health and pharmaceutical industry. A yeast Rhodotorula glutinis X-20 we obtained before has good selenium-enriched ability, but its selenium content is still low for industrial application. In this study, strategies of process optimization and transport regulation of selenium were thus employed to further improve the cell growth and selenium enrichment. Through engineering phosphate transporters from Saccharomyces cerevisiae into R. glutinis X-20, the selenium content was increased by 21.1%. Through using mixed carbon culture (20 g L-1, glycerol: glucose 3:7), both biomass and selenium content were finally increased to 5.3 g L-1 and 5349.6 µg g-1 (cell dry weight, DWC), which were 1.14 folds and 6.77 folds compared to their original values, respectively. Our results indicate that high selenium-enrichment ability and biomass production can be achieved through combining process optimization and regulation of selenium transport.


Assuntos
Engenharia Metabólica/métodos , Fosfatos/metabolismo , Rhodotorula/genética , Saccharomyces cerevisiae/genética , Selênio/metabolismo , Transgenes , Transporte Biológico , Biomassa , Meios de Cultura/química , Meios de Cultura/farmacologia , Fermentação , Expressão Gênica , Glucose/química , Glucose/metabolismo , Glicerol/química , Glicerol/metabolismo , Proteínas de Transporte de Fosfato/genética , Proteínas de Transporte de Fosfato/metabolismo , Plasmídeos/química , Plasmídeos/metabolismo , Simportadores de Próton-Fosfato/genética , Simportadores de Próton-Fosfato/metabolismo , Rhodotorula/metabolismo , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Proteínas Cotransportadoras de Sódio-Fosfato Tipo III/genética , Proteínas Cotransportadoras de Sódio-Fosfato Tipo III/metabolismo
16.
J Agric Food Chem ; 67(31): 8590-8598, 2019 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-31287301

RESUMO

Patchoulol, a natural sesquiterpene compound, is widely used in perfumes and cosmetics. Several strategies were adopted to enhance patchoulol production in Saccharomyces cerevisiae: (i) farnesyl pyrophosphate (FPP) synthase and patchoulol synthase were fused to increase the utilization of FPP precursor; (ii) expression of the limiting genes of the mevalonate pathway was enhanced; (iii) squalene synthase was weakened by a glucose-inducible promoter of HXT1 (promoter for hexose transporter) to reduce metabolic flux from FPP to ergosterol; and (iv) farnesol biosynthesis was inhibited to decrease the consumption of FPP. Glucose was used to balance the trade-off between the competitive squalene and patchoulol pathways. The patchoulol production was 59.2 ± 0.7 mg/L in a shaken flask with a final production of 466.8 ± 12.3 mg/L (20.5 ± 0.5 mg/g dry cell weight) combined with fermentation optimization, which was 7.8-fold higher than the reported maximum production. The work significantly promoted the industrialization process of patchoulol production using biobased microbial platforms.


Assuntos
Engenharia Metabólica , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Sesquiterpenos/metabolismo , Fermentação , Ácido Mevalônico/metabolismo , Fosfatos de Poli-Isoprenil/metabolismo , Regiões Promotoras Genéticas , Saccharomyces cerevisiae/enzimologia , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Esqualeno/metabolismo
17.
Water Sci Technol ; 79(11): 2203-2210, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31318358

RESUMO

The present study assessed the carbohydrate and sugar production from Chlorella spp. biomass harvested from a field scale reactor simulating phycoremediation of swine wastewater. The microalgae biomass was mainly composed by (%): carbohydrates (41 ± 0.4), proteins (50 ± 0.4), and lipids (1.3 ± 0.5). The residual sugar present in the biomass was extracted via acid hydrolysis. Among different concentrations of sulfuric acid tested (i.e., 47, 94, 188, 281 and 563 mM), significantly higher sugar content was obtained with 188 mM (0.496 g-sugar g-1 microalgae-DW). The concentration of sugar present in the microalgae did not differ significantly between the biomasses harvested by either centrifugation or coagulation-flocculation. Two commercially available strains of yeast (i.e., Saccharomyces cerevisiae and S. cerevisiae chardonnay) were tested for their capability to ferment sugar from lyophilized microalgae biomass. S. cerevisiae chardonnay showed a significantly faster consumption of sugar during the exponential growth phase. Both strains of yeast were capable of consuming most of the sugar added ≅ 8 g L-1 within 24 h. Overall, the results suggest that carbohydrate-rich microalgae biomass obtained from the phycoremediation of swine wastewaters can play an important role in green design for industries seeking alternative sources of feedstock rich in sugar.


Assuntos
Chlorella , Microalgas , Eliminação de Resíduos Líquidos , Animais , Biomassa , Carboidratos , Saccharomyces cerevisiae , Açúcares , Suínos , Águas Residuárias
18.
Sheng Wu Gong Cheng Xue Bao ; 35(7): 1214-1221, 2019 Jul 25.
Artigo em Chinês | MEDLINE | ID: mdl-31328478

RESUMO

Yeast cell wall plays an important role in the establishment and maintenance of cell morphology upon the cell wall stress. The cell wall of yeast consists of ß-glucans, mannoproteins and chitin. The composition and structure remodel due to cell wall stress. Brewer's yeast cell wall exhibits stress response during long-term acclimation in order to adapt to environmental changes. This paper reviews the composition and structure of yeast cell wall and the molecular mechanisms of cell wall remodeling and signal pathway regulation.


Assuntos
Parede Celular , Saccharomyces cerevisiae , Quitina
19.
Sheng Wu Gong Cheng Xue Bao ; 35(7): 1256-1265, 2019 Jul 25.
Artigo em Chinês | MEDLINE | ID: mdl-31328482

RESUMO

Naringenin is a natural flavonoid compound with anti-inflammatory, anti-oxidation, anti-viral, anti-atherosclerosis and other pharmacological activities. It is also an important precursor of other flavonoid synthesis and with great value of application. At present, the production of flavonoids such as naringenin by microbial methods has a low yield due to imbalance of metabolic pathways, which greatly limits its industrial application. In this study, a naringenin-producing strain of Saccharomyces cerevisiae Y-01 was used in the research object. The expression levels of 4-coumaric acid: CoA ligase (4CL), chalcone synthase (CHS) and chalcone isomerase (CHI) were controlled by promoter and copy numbers to investigate the quantitative effect of key enzyme expression level on the accumulation level of target products. The results showed that the correlation between naringenin production and 4CL or CHI expression was not significant while there was a positive correlation with the expression level of CHS. Strain Y-04 with high yield of naringenin was obtained by regulating the expression level of chs gene, and the yield was increased by 4.1-folds compared with the original strain Y-01. This study indicated that CHS is a key regulatory target of naringenin synthesis. Rational regulation of CHS expression can significantly promote the accumulation of naringenin. The related results provide an important theoretical reference for the use of metabolic engineering to strengthen microbial synthesis of important flavonoids such as naringenin.


Assuntos
Flavanonas/metabolismo , Engenharia Metabólica , Saccharomyces cerevisiae
20.
J Agric Food Chem ; 67(28): 7942-7953, 2019 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-31264861

RESUMO

Tryptophan, phenylalanine, and tyrosine play an important role as nitrogen sources in yeast metabolism. They regulate biomass production and fermentation rate, and their catabolites contribute to wine health benefits and sensorial character through the yeast biotransformation of grape juice constitutes into biologically active and flavor-impacting components. A UHPLC-MS/MS method was applied to monitor 37 tryptophan/phenylalanine/tyrosine yeast metabolites both in extra- and intracellular extracts produced by the fermentation of two Saccharomyces cerevisiae strains and one Torulaspora delbrueckii. The results shed light on the intra- and extra-cellular metabolomic dynamics, by combining metabolic needs, stimuli, and signals. Among others, the results indicated (a) the production of 2-aminoacetophenone by yeasts, mainly by the two Saccharomyces cerevisiae; (b) the deactivation and/or detoxification of tryptophol via sulfonation reaction; and (c) the deacetylation of N-acetyl tryptophan ethyl ester and N-acetyl tyrosine ethyl ester by producing the corresponding ethyl esters.


Assuntos
Aminoácidos Aromáticos/metabolismo , Saccharomyces cerevisiae/metabolismo , Torulaspora/metabolismo , Aminoácidos Aromáticos/química , Cromatografia Líquida de Alta Pressão , Nitrogênio/metabolismo , Saccharomyces cerevisiae/química , Espectrometria de Massas em Tandem , Torulaspora/química
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