The Essential Nucleolar Yeast Protein Nop8p Controls the Exosome Function during 60S Ribosomal Subunit Maturation

The yeast nucleolar protein Nop8p has previously been shown to interact with Nip7p and to be required for 60S ribosomal subunit formation. Although depletion of Nop8p in yeast cells leads to premature degradation of rRNAs, the biochemical mechanism responsible for this phenotype is still not known. In this work, we show that the Nop8p amino-terminal region mediates interaction with the 5.8S rRNA, while its carboxyl-terminal portion interacts with Nip7p and can partially complement the growth defect of the conditional mutant strain Dnop8/GAL:NOP8. Interestingly, Nop8p mediates association of Nip7p to pre-ribosomal particles. Nop8p also interacts with the exosome subunit Rrp6p and inhibits the complex activity in vitro, suggesting that the decrease in 60S ribosomal subunit levels detected upon depletion of Nop8p may result from degradation of pre-rRNAs by the exosome. These results strongly indicate that Nop8p may control the exosome function during pre-rRNA processing.

Aging and calorie restriction modulate yeast redox state, oxidized protein removal, and the ubiquitin-proteasome system

The ubiquitin-proteasome system governs the half-life of most cellular proteins. Calorie restriction (CR) extends the maximum life span of a variety of species and prevents oxidized protein accumulation. We studied the effects of CR on the ubiquitin-proteasome system and protein turnover in aging Saccharomyces cerevisiae. CR increased chronological life span as well as proteasome activity compared to control cells. The levels of protein carbonyls, a marker of protein oxidation, and those of polyubiquitinated proteins were modulated by CR. Controls, but not CR cells, exhibited a significant increase in oxidized proteins. In keeping with decreased proteasome activity, polyubiquitinated proteins were increased in young control cells compared to time-matched CR cells, but were profoundly decreased in aged control cells despite decreased proteasomal activity. This finding is related to a decreased polyubiquitination ability due to the impairment of the ubiquitin-activating enzyme in aged control cells, probably related to a more oxidative microenvironment. CR preserves the ubiquitin-proteasome system activity. Overall, we found that aging and CR modulate many aspects of protein modification and turnover. (C) 2011 Elsevier Inc. All rights reserved.

Ethanol production by a new pentose-fermenting yeast strain, Scheffersomyces stipitis UFMG-IMH 43.2, isolated from the Brazilian forest

The ability of a recently isolated Scheffersomyces stipitis strain (UFMG-IMH 43.2) to produce ethanol from xylose was evaluated. For the assays, a hemicellulosic hydrolysate produced by dilute acid hydrolysis of sugarcane bagasse was used as the fermentation medium. Initially, the necessity of adding nutrients (MgSO(4).7H(2)O, yeast extract and/or urea) to this medium was verified, and the yeast extract supplementation favoured ethanol production by the yeast. Then, in a second stage, assays under different initial xylose and cell concentrations, supplemented or not with yeast extract, were performed. All these three variables showed significant (p < 0.05) influence on ethanol production. The best results (ethanol yield and productivity of 0.19 g/g and 0.13 g/l/h, respectively) were obtained using the hydrolysate containing an initial xylose concentration of 30 g/l, supplemented with 5.0 g/l yeast extract and inoculated with an initial cell concentration of 2.0 g/l. S. stipitis UFMG-IMH 43.2 was demonstrated to be a yeast strain with potential for use in xylose conversion to ethanol. The establishment of the best fermentation conditions was also proved to be of great importance to increasing the product formation by this yeast strain. These findings open up new perspectives for the establishment of a feasible technology for ethanol production from hemicellulosic hydrolysates. Copyright (C) 2011 John Wiley & Sons, Ltd.

Evaluation of hydrodynamic parameters of a fluidized-bed reactor with immobilized yeast

The fluidized bed reactor has successfully been used to perform biotechnological processes addressed to the production of high added value. The present work evaluates hydrodynamic parameters of a bench-scale fluidized bed reactor with cells of the yeast Candida guilliermondii immobilized either in calcium alginate beads or in polyvinyl alcohol (PVA). The effects of the following variables on cell immobilization were evaluated at 30 degrees C and feeding a synthetic medium containing 50 g L-1 xylose: total particle density (cells plus support), terminal velocity, particle drag force, minimum fluidization velocity and bed porosity. According to the results obtained, the reactor was shown to operate like a fixed-bed bioreactor at xi < 0.5 and a fluidized bed bioreactor at xi > 0.5. The maximum flow rate needed to obtain maximum bed fluidization in the reactor was equal to the terminal velocity of the immobilized cell particles. Particles of cells immobilized within these supports showed values of drag coefficient lower than those reported for other high-density supports. The evaluation of these hydrodynamic characteristics lead to an adequate bed fluidization inside the reactor, thus improving oxygen transference and availability in the fermentation medium, making the process more viable for future scale-up. (c) 2008 Society of Chemical Industry.

Biological detoxification of different hemicellulosic hydrolysates using Issatchenkia occidentalis CCTCC M 206097 yeast

This work had as its main objective to contribute to the development of a biological detoxification of hemicellulose hydrolysates obtained from different biomass plants using Issatchenkia occidentalis CCTCC M 206097 yeast. Tests with hemicellulosic hydrolysate of sugarcane bagasse in different concentrations were carried out to evaluate the influence of the hydrolysate concentration on the inhibitory compounds removal from the sugarcane bagasse hydrolysate, without reduction of sugar concentration. The highest reduction values of inhibitors concentration and less sugar losses were observed when the fivefold concentrated hydrolysate was treated by the evaluated yeast. In these experiments it was found that the high sugar concentrations favored lower sugar consumption by the yeast. The highest concentration reduction of syringaldehyde (66.67%), ferulic acid (73.33%), furfural (62%), and 5-HMF (85%) was observed when the concentrated hydrolysate was detoxified by using this yeast strain after 24 h of experimentation. The results obtained in this work showed the potential of the yeast Issatchenkia occidentalis CCTCC M 206097 as detoxification agent of hemicellulosic hydrolysate of different biomass plants.

Raspberry (Rubus idaeus L.) wine: Yeast selection, sensory evaluation and instrumental analysis of volatile and other compounds

To evaluate the potential for fermentation of raspberry pulp, sixteen yeast strains (S. cerevisiae and S. bayanus) were studied. Volatile compounds were determined by GC-MS, GC-FID, and GC-PFPD. Ethanol. glycerol and organic acids were determined by HPLC. HPLC-DAD was used to analyse phenolic acids. Sensory analysis was performed by trained panellists. After a screening step, CAT-1, UFLA FW 15 and S. bayanus CBS 1505 were previously selected based on their fermentative characteristics and profile of the metabolites identified. The beverage produced with CAT-1 showed the highest volatile fatty acid concentration (1542.6 mu g/L), whereas the beverage produced with UFLA FIN 15 showed the highest concentration of acetates (2211.1 mu g/L) and total volatile compounds (5835 mu g/L). For volatile sulphur compounds. 566.5 mu g/L were found in the beverage produced with S. bayanus CBS 1505. The lowest concentration of volatile sulphur compounds (151.9 mu g/L) was found for the beverage produced with UFLA FW 15. In the sensory analysis, the beverage produced with UFLA FW 15 was characterised by the descriptors raspberry, cherry, sweet, strawberry, floral and violet. In conclusion, strain UFLA FW 15 was the yeast that produced a raspberry wine with a good chemical and sensory quality. (C) 2010 Elsevier Ltd. All rights reserved.

A comparison of the critical impeller speed for solids suspension in a bench-scale and a pilot-scale mechanical flotation cell

This paper compares the critical impeller speed results for 6 L Denver and Wemco bench-scale flotation cells with findings from a study by Van der Westhuizen and Deglon [Van der Westhuizen, A.P., Deglon, D.A., 2007. Evaluation of solids suspension in a pilot-scale mechanical flotation cell: the critical impeller speed. Minerals Engineering 20,233-240; Van der Westhuizen, A.P., Deglon, D.A., 2008. Solids suspension in a pilot scale mechanical flotation cell: a critical impeller speed correlation. Minerals Engineering 21, 621-629] conducted in a 125 L Batequip flotation cell. Understanding solids suspension has become increasingly important due to dramatic increases in flotation cell sizes. The critical impeller speed is commonly used to indicate the effectiveness of solids suspension. The minerals used in this study were apatite, quartz and hematite. The critical impeller speed was found to be strongly dependent on particle size, solids density and air flow rate, with solids concentration having a lesser influence. Liquid viscosity was found to have a negligible effect. The general Zwietering-type critical impeller speed correlation developed by Van der Westhuizen and Deglon [Van der Westhuizen, A.P., Deglon, D.A., 2008. Solids suspension in a pilot scale mechanical flotation cell: a critical impeller speed correlation. Minerals Engineering 21, 621-629] was found to be applicable to all three flotation machines. The exponents for particle size, solids concentration and liquid viscosity were equivalent for all three cells. The exponent for solids density was found to be less significant than that obtained by the previous authors, and to be consistent with values reported in the general literature for stirred tanks. Finally, a new dimensionless critical impeller speed correlation is proposed where the particle size is divided by the impeller diameter. This modified equation generally predicts the experimental measurements well, with most predictions within 10% of the experimental. (C) 2009 Elsevier Ltd. All rights reserved.

Froth flotation of coal tailings

The processes involved in coal preparation produce harmful effects for the environment, mainly by the quantity and nature Of wastes that are generated, These tailings generally called ""pyritic"", even though not presenting a high pyrite content, are an acid material harmful to the environment. These tailings are usually disposed of in tailings dams. The tailings, studied in this work come from a process involving dense-medium cyclones and spirals. They have an ash content of 56% and a calorific value of 5,800 BTU/Lb, the sulfur content is 1.2%. In terms of size, the material is considered as ultrafine, as 63% of it is less than 0,014 mm. The coal matter content of these tailings is easily recovered by froth flotation, as evidenced in this work. It possible to recover 74% of the coal matter and to obtain a product with 7.3% ash and calorific value of 14,225 BTU/lb in dry basis.

Suspension of apatite particles in a self-aerated Denver laboratory flotation cell

Effective solids suspension is a necessary precondition for particle collection, and solids suspension is largely dependent on the hydrodynamics of the flotation cell. This study attempted to correlate the status of the suspension of apatite particles of different sizes in a Denver laboratory flotation cell versus the impeller rotational speed (N) adopted to operate the machine. The latter variable (N) influences the impeller capacity to lift the particles from the bottom of the tank and also to disperse them throughout the volume of the vessel. Such an impeller capacity can be characterized by the critical impeller speed for the accomplishment of solids off-bottom suspension (N(z)) and also by the velocity of the radial water flow discharged by the impeller (U) divided by the particle terminal settling velocity (U(s)). This way, the status of the suspension of apatite particles inside the flotation cell can be characterized by one of three categories: ""segregation"" (N/N(2) < 0.60 and U(s)/U > 0.08); ""suspension"" (0.60 <= N/N(2) < 1 and 0.06 < U(s)/U < 0.10); and ""dragging"" (N/N(2) >= 1 and U(s)/U <= 0.03). The range of impeller rotational speed (N), which was able to suspend the finest particles (D(p) = 90,mu m), was unable to suspend the coarsest particles (D(P) = 254 mu m). Conversely, the high value of N (N > 1,300 rpm), which is adequate to suspend the coarsest particles, may promote the entrainment of the finest particles to the froth layer.

Surface tension of flotation solution and its influence on the selectivity of the separation between apatite and gangue minerals

This work presents and discusses the influence of the surface tension (gamma(LV)) of methanol/water mixtures on the flotation response of apatite versus gangue minerals conditioned with flotation reagents (75 g/t cornstarch and 100 g/t Berol 867) at pH 10.6. Berol 867 is a collector composed of sodium alkyl sarcosinate plus nonionic surfactant. The highest Schulz efficiency of separation (recovery of apatite minus recovery of gangue) was achieved at approximate to 51.0 mN/m. The critical surface tension of wettability (gamma(C)) of apatite was found to occur at 34.7 mN/m when determined by means of gamma flotation experiments, , and it was 33.9 mN/m when determined by Zisman`s approach.

Separation of silica from bauxite via froth flotation

This paper reports an innovative development: concentrating gibbsite via reverse froth flotation in order to obtain a metallurgical-grade bauxite concentrate. Tailings from an industrial plant have undergone attrition scrubbing and desliming; the quartz silica contained in the tailings has undergone flotation. Starch was used as a depressant, and ether-amine as the cationic collector. Optimum pH is around 10.0. In pilot plant scale, a metallurgical-grade concentrate was obtained by assaying 42.3% available alumina with an alumina/insoluble silica mass ratio of 11.1. It contained the gibbsite and the iron and titanium bearing minerals. The concentrate was further upgraded by magnetic separation, leading to 54.0% available alumina, with an alumina/insoluble silica mass ratio of 12.6 at an overall available alumina recovery of 69.3% in the final concentrate (non-magnetic product). (C) 2008 Elsevier Ltd. All rights reserved.

Considerations on the kinetics of froth flotation of ultrafine coal contained in tailings

This article presents a kinetic evaluation of froth flotation of ultrafine coal contained in the tailings from a Colombian coal preparation plant. The plant utilizes a dense-medium cyclones and spirals circuit. The tailings contained material that was 63% finer than 14 mu m. Flotation tests were performed with and without coal ""promoters"" (diesel oil or kerosene) to evaluate the kinetics of flotation of coal. It was found that flotation rates were higher when no promoter was added. Different kinetic models were evaluated for the flotation of the coal from the tailings, and it was found that the best fitted model was the classical first-order model.

Mitochondrial function in the yeast form of the pathogenic fungus Paracoccidioides brasiliensis

Differences between the respiratory chain of the fungus Paracoccidioides brasiliensis and its mammalian host are reported. Respiration, membrane potential, and oxidative phosphorylation in mitochondria from P. brasiliensis spheroplasts were evaluated in situ, and the presence of a complete (Complex I-V) functional respiratory chain was demonstrated. In succinate-energized mitochondria, ADP induced a transition from resting to phosphorylating respiration. The presence of an alternative NADH-ubiquinone oxidoreductase was indicated by: (i) the ability to oxidize exogenous NADH and (ii) the lack of sensitivity to rotenone and presence of sensitivity to flavone. Malate/NAD(+)-supported respiration suggested the presence of either a mitochondrial pyridine transporter or a glyoxylate pathway contributing to NADH and/or succinate production. Partial sensitivity of NADH/succinate-supported respiration to antimycin A and cyanide, as well as sensitivity to benzohydroxamic acids, suggested the presence of an alternative oxidase in the yeast form of the fungus. An increase in activity and gene expression of the alternative NADH dehydrogenase throughout the yeast`s exponential growth phase was observed. This increase was coupled with a decrease in Complex I activity and gene expression of its subunit 6. These results support the existence of alternative respiratory chain pathways in addition to Complex I, as well as the utilization of NADH-linked substrates by P. brasiliensis. These specific components of the respiratory chain could be useful for further research and development of pharmacological agents against the fungus.

Involvement of an Alternative Oxidase in Oxidative Stress and Mycelium-to-Yeast Differentiation in Paracoccidioides brasiliensis

Paracoccidioides brasiliensis is a thermodimorphic human pathogenic fungus that causes paracoccidioidomycosis (PCM), which is the most prevalent systemic mycosis in Latin America. Differentiation from the mycelial to the yeast form (M-to-Y) is an essential step for the establishment of PCM. We evaluated the involvement of mitochondria and intracellular oxidative stress in M-to-Y differentiation. M-to-Y transition was delayed by the inhibition of mitochondrial complexes III and IV or alternative oxidase (AOX) and was blocked by the association of AOX with complex III or IV inhibitors. The expression of P. brasiliensis aox (Pbaox) was developmentally regulated through M-to-Y differentiation, wherein the highest levels were achieved in the first 24 h and during the yeast exponential growth phase; Pbaox was upregulated by oxidative stress. Pbaox was cloned, and its heterologous expression conferred cyanide-resistant respiration in Saccharomyces cerevisiae and Escherichia coli and reduced oxidative stress in S. cerevisiae cells. These results reinforce the role of PbAOX in intracellular redox balancing and demonstrate its involvement, as well as that of other components of the mitochondrial respiratory chain complexes, in the early stages of the M-to-Y differentiation of P. brasiliensis.

Spray Drying of Extracts from Red Yeast Fermentation Broth

Red yeast rice is a pigmented material that is traditionally used in Asia as a food colorant. In addition to food applications, red yeast rice is known in traditional Chinese medicine for its therapeutic actions. The aim of this work was to study the quality interactions during spray drying of extracts from the Monascus ruber van Tiegham fermentation broth. The quality indicators used for the dry powder properties were the levels of monacolin K, ratio of red to yellow pigments, as well as their antioxidant activity. The experiments followed a Box-Behnken design to study the effects of the adjuvant/drug ratio, adjuvant incorporation time, and oulet drying temperature on the pharmacotechnical, chemical, and biological properties of the dry extract. The influences of these factors on the characteristics of the dry powder were evaluated by the bulk density, tapped density, Carr index, Hausner factor, residual moisture content, water activity, antioxidant activity, monacolin K, yellow-to-red pigment ratio, and antioxidant activity. The analysis of variance (ANOVA) on experimental data revealed that an increase in drying temperature significantly increased the dry powder yield and caused an improvement in powder flow properties, which may be related to lower moisture contents. The drying temperature did not affect the monacolin K content in dry powder but showed a complex influence on its antioxidant activity. The increase in drying adjuvant-to-drug ratio affected the yield and also indicated a protective effect on the monacolin K content. The duration of drying adjuvant incorporation had little or negligible effect on powder properties. The dry extracts of red yeast rice showed adequate properties and the process proposed herein can be used to prepare nutraceutical products.