Yeast Cell Wall Fractions Research Articles

Mitigation of AFB1-Related Toxic Damage to the Intestinal Epithelium in Broiler Chickens Consumed a Yeast Cell Wall Fraction.

In vivo experiments were conducted to evaluate the effectiveness of a yeast cell wall fraction (YCW) to reduce the negative impact of aflatoxin B1 (AFB1) to the intestinal epithelium in broiler chickens. Zeta potential (ζ-potential), point of zero charge (pHpzc), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM) techniques were used to characterize the YCW. Two hundred one-day-old male Ross 308 broiler chickens were randomly allocated into four treatments: (1) control, chickens fed an AFB1-free diet; (2) AF, chickens feed an AFB1-contaminated diet (500 ng AFB1/g); (3) YCW, chickens fed an AFB1-free diet + 0.05% YCW; and (4) AF + YCW, chickens fed an AFB1-contaminated diet (500 ng AFB1/g) + 0.05% YCW. At the end of the 21-day feeding period, fluorescein isothiocyanate dextran (FITC-d) was administered to chicks by oral gavage to evaluate gastrointestinal leakage. Blood and duodenum samples were collected to assess serum biochemistry and histomorphology, respectively. Compared to the control group, chicks of the AF group significantly diminished weight gain (WG) and average daily feed intake (ADFI), and increased feed conversion ratio (FCR), mortality rate (MR), and intestinal lesion scores (p < 0.05). Alterations in some serum biochemical parameters, and damage to the intestinal integrity were also evident in the AF-intoxicated birds. YCW supplementation improved WG and FCR and increased villus height, villus area, crypt depth, and the number of goblet cells in villi. The effects of YCW on growth performance were not significant in chicks of the AF + YCW group; however, the treatment decreased MR and significantly ameliorated some biochemical and histomorphological alterations. The beneficial effect of YCW was more evident in promoting gut health since chickens of the AF + YCW group presented a significant reduction in serum FITC-d concentration. This positive effect was mainly related to the changes in negative charges of YCW due to changes in pH, the net negative surface charge above the pHpzc, the higher quantities of negative charged functional groups on the YCW surface, and its ability to form large aggregates. From these results, it can be concluded that YCW at low supplementation level can partially protect broilers' intestinal health from chronic exposure to AFB1.

Supplementation of Yeast Cell Wall Fraction Tends to Improve Intestinal Health in Adult Dogs Undergoing an Abrupt Diet Transition.

When owners decide to change their pet's food, a rapid transition may cause gastrointestinal distress. Yeast products may help with digestive upset during diet transition due to the bioactive compounds they possess, which may lead to improved intestinal morphology and integrity, modified gut microbiota, and modulated immune responses. The objective of this study was to determine the effects of a yeast cell wall fraction supplement on measures of gut integrity and fecal characteristics of adult dogs undergoing an abrupt diet transition. Twelve adult female beagles (mean age: 5.16 ± 0.87 years; mean body weight: 13.37 ± 0.68 kg) were used in a replicated 4 × 4 Latin square design with four 28-day experimental periods. During days 1–14, dogs were fed a dry kibble diet and supplemented with a placebo (cellulose; 125 mg/d) or yeast product (365 mg/d; equivalent to 0.2% of diet). During days 15–28, dogs remained on their placebo or yeast treatments but were rapidly transitioned to a canned diet or high-fiber diet. Fresh fecal samples were collected on days 13, 16, 20, 24, and 27 for measurement of pH, dry matter, calprotectin, immunoglobulin A (IgA), Escherichia coli, and Clostridium perfringens. Blood samples were collected on days 14, 17, and 28 to measure serum lipopolysaccharide-binding protein concentrations. All data were analyzed using the Mixed Models procedure of SAS 9.4. Fecal pH, dry matter, calprotectin, IgA, and E. coli were not affected (P > 0.05) by treatment before diet transition. Dogs supplemented with yeast cell wall fraction tended to have higher (P = 0.06) fecal C. perfringens counts than the controls. After diet transition, most parameters were not altered (P > 0.05) by treatment except that yeast-supplemented dogs tended to have higher (P = 0.06) fecal IgA than controls. Our results suggest that the yeast product may modestly improve intestinal health after an abrupt diet transition in adult dogs by enhancing intestinal immunity.

Equilibrium Isotherm Approach to Measure the Capability of Yeast Cell Wall to Adsorb Clostridium perfringens.

Yeast cell wall (YCW) products are currently used as substitutes to antibiotic growth promoters, to improve animal performances, and to reduce the incidence of infectious diseases in livestock. They are claimed to bind enteropathogens, thus interfering with their colonization in the intestinal mucosa. Although the anti-infectious activity of YCW products on Gram-positive pathogens like Clostridium perfringens has been reported in vivo, in vitro evidences on the adsorption of C. perfringens by YCW fractions are not yet available. Preliminary results showed that purified YCW products exert antimicrobial activity toward C. perfringens. Using the adsorption isotherm approach, we measured the ability of YCW products in adsorbing C. perfringens, thus reducing its viability. Dosages of YCW products >1 mg/mL adsorbed 4 Log colony-forming unit (CFU)/mL of C. perfringens in buffered solution. The maximum adsorption of the bacterium was reached in 3 h, whereas only one product of four YCW products retained the adsorption up to 6 h. The analysis of equilibrium isotherms and adsorption kinetics revealed that all products adsorb C. perfringens in a dose- and time-dependent manner, with high affinity and capacity, sequestering up to 4 Log CFU/mg of product. The determination of adsorption parameters allows to differentiate among adsorbents and select the most efficient product. This approach discriminated among YCW products more efficiently than the antimicrobial assay. In conclusion, this study suggests that the ability of YCW products in reducing C. perfringens viability can be the result of an adsorption mechanism.

Yeast derived from lignocellulosic biomass as a sustainable feed resource for use in aquaculture.

The global expansion in aquaculture production implies an emerging need of suitable and sustainable protein sources. Currently, the fish feed industry is dependent on high-quality protein sources of marine and plant origin. Yeast derived from processing of low-value and non-food lignocellulosic biomass is a potential sustainable source of protein in fish diets. Following enzymatic hydrolysis, the hexose and pentose sugars of lignocellulosic substrates and supplementary nutrients can be converted into protein-rich yeast biomass by fermentation. Studies have shown that yeasts such as Saccharomyces cerevisiae, Candida utilis and Kluyveromyces marxianus have favourable amino acid composition and excellent properties as protein sources in diets for fish, including carnivorous species such as Atlantic salmon and rainbow trout. Suitable downstream processing of the biomass to disrupt cell walls is required to secure high nutrient digestibility. A number of studies have shown various immunological and health benefits from feeding fish low levels of yeast and yeast-derived cell wall fractions. This review summarises current literature on the potential of yeast from lignocellulosic biomass as an alternative protein source for the aquaculture industry. It is concluded that further research and development within yeast production can be important to secure the future sustainability and economic viability of intensive aquaculture. © 2016 Society of Chemical Industry.

Quantitative in vitro assay to evaluate the capability of yeast cell wall fractions from Trichosporon mycotoxinivorans to selectively bind gram negative pathogens

Yeast cell wall fractions have been proposed to bind enteropathogenic bacteria. The aim of this study was to develop a quantitative assay by measuring the optical density as growth parameter of adhering bacteria. The exponential growth phase of adhering bacteria was determined by optical density reading and compared with the colony count (CFU/mL). A linear regression was compiled and the bacterial number bound to the yeast cell wall product could be determined. Further focus was the investigation of a yeast cell wall from strain Trichosporon mycotoxinivorans (MTV) for its ability to bind gram negative Salmonella, E. coli and Campylobacter strains and gram positive probiotic bacteria of the genera lactobacilli and bifidobacteria as well as gram positive Clostridium perfringens quantitatively. The gram negative probiotic strain E. coli Nissle 1917 was also investigated. Seven out of 10 S. Typhimurium and S. Enteritidis strains adhered to the cell wall product with an amount between 10 3 and 10 4 CFU/10 μg. Four out of 7 E. coli strains showed an average binding capability (10 2 CFU/10 µg) whereas 4 × 10 3 E. coli F4 cells bound per 10 μg yeast cell wall. E. coli 0149 K91, E. coli 0147 K89, C. jejuni and C. perfringens as well the genera lactobacilli and bifidobacteria did not bind to the yeast cell wall. E. coli Nissle 1917 was bound with 2 × 10 2 CFU/10 μg. These results demonstrate that cell wall from MTV can be used to differentially bind E. coli spp. and Salmonella spp. up to 8 × 10 4 CFU/10 μg. Thus certain yeast cell walls may prevent enteric infections caused by selective bacteria. This methodical approach would be an accurate tool in the feed industry for quality control of yeast cell wall products.

Determination of Reduced Cysteine in Oenological Cell Wall Fractions of Saccharomyces cerevisiae

Compounds containing cysteine residues, such as glutathione, can affect the redox potential of must and wine by reduction of o-quinones and hydrogen peroxide. The oenological yeast cell wall fractions contain cysteine residues in their protein structure, and they could affect both oxidative and odor properties of wine. An analytical approach based on the derivatization of cysteinyl residues with p-benzoquinone followed by reversed-phase high-performance liquid chromatography separation was developed to quantify glutathione and free and protein cysteine in 16 Saccharomyces cerevisiae strains and 12 commercial samples of yeast mannoproteins, hulls, and lysates. The chemical modifications induced by the Maillard reaction following the industrial preparation of such fractions were evaluated as well. Lysates showed the highest protein cysteine content and high contents of glutathione and free cysteine. Mannoproteins showed an intense Maillard reaction (furosine >60 mg/100 g protein), and most of the samples were able to bind thiol compounds with a potentially detrimental effect toward the thiol-related odors in wine.

Influência de frações da parede celular de levedura (Saccharomyces cerevisiae) sobre os índices séricos de glicose e lipídios, microbiota intestinal e produção de ácidos graxos voláteis (AGV) de cadeias curtas de ratos em crescimento

Os índices séricos de glicose e lipídios, a microbiota intestinal e a produção de ácidos graxos voláteis de cadeias curtas (AGV) foram determinados em ratos Wistar submetidos às dietas: padrão (AIN-P), padrão modificada (AIN-M) e às dietas contendo frações de parede celular de levedura: glicana insolúvel (GI), manana (M) e glicana mais manana (G+M), como única fonte de fibra alimentar. O fracionamento da parede celular (PC) foi realizado por processos físicos e químicos de extração, centrifugação e secagem em "spray dryer". Os índices séricos foram dosados através de "kits" comerciais. A microbiota e a produção de AGV foram determinadas nos conteúdos intestinais, incluindo cólon, ceco e reto. Considerando os níveis de colesterol no tempo (T0) e no tempo 28 (T28), as dietas AIN-P, AIN-M e M apresentaram efeito hipocolesterolêmico, tendo em vista que a composição das dietas eram de natureza hipercolesterolêmica. Em relação à glicose sérica, no tempo (T0) observou-se uma elevação geral da glicemia, sugerindo um efeito hiperglicêmico das dietas estudadas. A dieta G+M foi a que apresentou valores significantemente mais elevados de lipídios séricos no tempo T14, e os níveis mais baixos foram observados na dieta M e na dieta GI no T14 e nas dietas AIN-M e AIN-P. A dieta AIN-P foi a que apresentou valor significantemente mais elevado de triacilgliceróis nos tempos T14 e T28. Os níveis mais baixos nos tempos T14 foram constatados para as dietas G+M e GI e no tempo T28 para as dietas AIN-M e M. De um modo geral, não houve modificações significativas na microbiota intestinal dos animais em nenhuma das dietas. Dentre os AGV, o ácido acético foi o predominante, seguido do propiônico e do butírico, em todas as dietas estudadas.

Isolation and Characterization of a Thermally Extracted Yeast Cell Wall Fraction Potentially Useful for Improving the Foaming Properties of Sparkling Wines

Two different yeast cell wall extracts were obtained using enzymatic digestion and thermal treatment. The effects of the extracts obtained on the foaming properties of a model wine and two sparkling wines were studied. The model wine and sparkling wines, supplemented with the thermal extract, presented better foaming properties than did the samples supplemented with the enzymatic extract. The fractioning (Con A chromatography) and characterization (SDS-PAGE, SEC, GC, and RP-HPLC) of both extracts showed that the fraction responsible for the foaming properties is constituted by mannoproteins with a relative molecular weight between 10 and 30 kDa, presenting an equilibrated composition of the hydrophobic and hydrophilic protein domains. This thermal extract did not modify the protein stability in both the model wine and the sparkling wines. These results demonstrate that the enrichment of a sparkling wine with mannoproteins extracted by mild heat procedures will contribute to improving its foaming properties.

Construction of a Library of Human Glycosyltransferases Immobilized in the Cell Wall ofSaccharomyces cerevisiae

Fifty-one human glycosyltransferases were expressed in Saccharomyces cerevisiae as immobilized enzymes and were assayed for enzymatic activities. The stem and catalytic regions of sialyl-, fucosyl-, galactosyl-, N-acetylgalactosaminyl-, and N-acetylglucosaminyltransferases were fused with yeast cell wall Pir proteins, which anchor glycosyltransferases at the yeast cell wall glucan. More than 75% of expressed recombinant glycosyltransferases retained their enzymatic activities in the yeast cell wall fraction and will be used as a human glycosyltransferase library. In increasing the enzymatic activities of immobilized glycosyltransferases, several approaches were found to be effective. Additional expression of yeast protein disulfide isomerase increased the expression levels and activities of polypeptide N-acetylgalactosaminyltransferases and other glycosyltransferases. PIR3 and/or PIR4 was more effective than PIR1 as a cell wall anchor when the Pir-glycosyltransferase fusions were expressed under the control of the constitutive glyceraldehyde-3-phosphate dehydrogenase promoter. Oligosaccharides such as Lewis x, Lewis y, and H antigen were successfully synthesized using this immobilized glycosyltransferase library, indicating that the Pir-fused glycosyltransferases are useful for the production of various human oligosaccharides.

In vitro biosorption of ochratoxin A on the yeast industry by-products: Comparison of isotherm models

Biosorption of ochratoxin A (OA) onto yeast biomass appears to be a reasonably low cost decontamination method. In vitro adsorption of OA onto three yeast industry by-products: a vinasse containing yeast cell walls (EX16), a purified yeast beta-glucan (BETA) and a yeast cell wall fraction (LEC) was examined at 25 °C. Seven classical adsorption models were tested to provide the best description of toxin adsorption. A comparison of these models was performed using the magnitude of the coefficient of determination R 2 for the linear models and the value of the sum of normalised errors (SNE) for linear and non-linear models. Based on the R 2 and the SNE values, Hill, Freundlich and Brunauer–Emmett–Teller equations produced the best models for OA biosorption onto respectively, EX16, BETA and LEC. For these best models, the values of isotherm constants were consistent when measured using both linear and non-linear calculations. The SNE calculation procedure presented in this paper in association with the linear equation analysis method is an appropriate approach for designing a better adsorption isothermal model.

Effect of temperature on in vitro ochratoxin A biosorption onto yeast cell wall derivatives

In vitro biosorption of ochratoxin A (OA) onto three yeast industry products: a vinasse containing yeast cell walls (EX16), a purified yeast beta glucan (BETA) and a yeast cell wall fraction (LEC), has been studied as a function of the temperature. Equilibrium binding assays were performed from 4 to 37 °C. The best models for OA biosorption onto EX16, BETA and LEC were identified as respectively Hill's, Freundlich and Brunauer-Emmett-Teller models. LEC was the most effective adsorbent for OA removal in aqueous solutions. All biosorption processes were spontaneous (negative Δ G 0) and exothermic (negative Δ H 0). For EX16 and BETA, the favourable enthalpic contributions (negative Δ H 0 values) to OA binding, were associated to unfavourable entropic contributions (negative Δ S 0 values). By contrast, the entropic contribution to OA binding was favourable (positive Δ S 0 values) in the case of LEC. The calculated values of the heat capacity (ΔCp) were close to zero, indicating the absence of a temperature dependency for Δ H and Δ S. The results suggest that the remarkable OA biosorption onto LEC involves both polar and non-polar non-covalent interactions and the concomitant reorganization of the water molecules of the solvent.

Preparation of spent brewer's yeast beta-glucans with a potential application as an immunostimulant for black tiger shrimp, Penaeus monodon.

Spent brewer's yeast was autolysed and the insoluble yeast cell wall fraction obtained was used as a raw material for the preparation of brewer's yeast beta-glucan (BYG). A simple alkaline extraction was applied and optimized. The BYG with significantly (P</=0.05) high carbohydrate and glucan contents of 92% and 51% (w/w), respectively and with a low protein content of 1.6% (w/w) was obtained by a single alkaline extraction using 5 volumes of 1.0 N NaOH at 90 degrees C for 1 h. In vitro, this glucan significantly (P</=0.05) enhanced phenoloxidase (PO) activity of black tiger shrimp hemolymph as compared to controls without added glucan. Also in vivo, an oral administration of 0.2% (w/w) in diets for 3 days significantly (P</=0.05) increased the PO-activity of the shrimp. The potential immunostimulating properties of a commercial and other yeast-derived products were also evaluated for comparative purposes.

Candidacidal Activity of Salivary Histatins: IDENTIFICATION OF A HISTATIN 5-BINDING PROTEIN ON Candida albicans

Candida albicans is the predominant species of yeast isolated from patients with oral candidiasis, which is frequently a symptom of human immunodeficiency virus infection and is a criterion for staging and progression of AIDS. Salivary histatins (Hsts) are potent in vitro antifungal agents and have great promise as therapeutic agents in humans with oral candidiasis. The molecular mechanisms by which Hsts kill yeast cells are not known. We report here, that unlike other antimicrobial proteins, Hsts do not display lytic activities to lipid membranes, measured by release and dequenching of the fluorescent dye calcein. Analysis of the magnitude and time course of Hst-induced calcein release from C. albicans cells further showed that loss of cell integrity was a secondary effect following cell death, rather than the result of primary disruption of the yeast cell membrane. 125I-Hst 5 binding studies indicated that C. albicans expressed a class of saturable binding sites (KD = 1 microM), numbering 8.6 x 10(5) sites/cell. Both Hst 3 and Hst 4 competed for these binding sites with similar affinities, which is consistent with the micromolar concentration of Hsts required for candidacidal activity. Specific 125I-Hst 5 binding was not detected to C. albicans spheroplasts, which were 14-fold less susceptible to Hst 5 killing, compared with intact cells in candidacidal assays. In overlay experiments, 125I-Hst 5 bound to a 67-kDa protein detected in C. albicans whole cell lysates and crude membrane fractions, but not in the yeast cell wall fraction. Consistent with the overlay data, cross-linking of 125I-Hst 5 to C. albicans resulted in the appearance of a specific 73-kDa 125I-Hst 5-containing complex that was not detected in the cell wall. 125I-Hst 5-binding protein of similar size was also observed in susceptible S. cerevisiae strain TI#20. This is the first description of Hst 5 binding sites on C. albicans which mediate cell killing and identification of a 67-kDa yeast Hst 5-binding protein. The binding characteristics of Hst 5 are in agreement with the observed potency of its biological effect and provide crucial information to the use of Hst 5 as a therapeutic agent. The presence of a specific C. albicans Hst 5-binding protein provides further insight into the potential mechanism of yeast killing and suggests a basis for differential activity between yeast killing and the nontoxic nature of Hsts to humans.

Yeast cell wall, membrane, and soluble marker polypeptides identified by comparative two-dimensional electrophoresis.

Yeast cell wall, plasma membrane, total spheroplast, and total soluble protein fractions were isolated from exponentially growing Saccharomyces cerevisiae batch cultures. The cell wall, plasma membrane, and soluble protein fractions were obtained by mechanical disruption of intact yeast cells under identical osmotic conditions. Electron micrographs of purified wall fractions appeared free of vesicular membrane contamination and micrographs of plasma membrane vesicles were free of cell wall contamination. Various stages of cell wall purification were monitored by electron microscopy and comparative two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This resulted in the identification of a glycopeptide designated 16w in the cell wall fraction, with an apparent isoelectric point of 5.0 and an apparent molecular weight of 25 000. Protein analyses of soluble and plasma membrane protein fractions failed to detect component 16w. Two-dimensional protein analyses of total cellular homogenates were capable of resolving the cell wall glycopeptide 16w. However, protein separations of spheroplasts formed by glusulase degradation of the cell wall complex did not detect 16w. These observations suggest that component 16w is unique to the cell wall fraction. In addition, comparison of two-dimensional gels of soluble and plasma membrane proteins, with a total cellular homogenate, tentatively identified several polypeptides unique to each of the soluble and plasma membrane fractions.