Ethanol Production by Fermentation Using Immobilized Cells of Saccharomyces cerevisiae in Cashew Apple Bagasse

In this work, cashew apple bagasse (CAB) was used for Saccharomyces cerevisiae immobilization. The support was prepared through a treatment with a solution of 3% HCl, and delignification with 2% NaOH was also conducted. Optical micrographs showed that high populations of yeast cells adhered to pre-treated CAB surface. Ten consecutive fermentations of cashew apple juice for ethanol production were carried out using immobilized yeasts. High ethanol productivity was observed from the third fermentation assay until the tenth fermentation. Ethanol concentrations (about 19.82–37.83 g L^?1 in average value) and ethanol productivities (about 3.30–6.31 g L^?1 h^?1) were high and stable, and residual sugar concentrations were low in almost all fermentations (around 3.00 g L^?1) with conversions ranging from 44.80% to 96.50%, showing efficiency (85.30–98.52%) and operational stability of the biocatalyst for ethanol fermentation. Results showed that cashew apple bagasse is an efficient support for cell immobilization aiming at ethanol production.     

Dry Red Wine Making Using Yeast Immobilized on Cork Pieces

The commercial Saccharomyces cerevisiae Uvaferme 299 wine yeast was immobilized on cork pieces to produce a biocatalyst for use in dry red wine making. The immobilized biocatalyst was suitable for clarified and non-clarified grape must fermentation at ambient and low temperatures (0–25 °C). Fermentation times were low and very low amounts of residual sugar were detected, showing suitability for dry wine production. The presence of suspended solids in the non-clarified must did not affect the activity of the immobilized cells. Complete fermentation of sugars at 0 °C was possible with immobilized Uvaferme 299, although not a cryotolerant strain, indicating a cryoprotective effect of cork. The presence of cork did not affect the composition of major volatiles with methanol and acetaldehyde kept at low levels. Reduction of amyl alcohols on total volatiles was also observed in wines fermented at low temperatures. Differences in the headspace aroma profile in wines produced by immobilized and free cells were found by GC–MS analysis, but no cork taint compounds were detected.     

Interfacial Engineering for Efficient Low-Temperature Flexible Perovskite Solar Cells

Photovoltaic technology with low weight, high specific power in cold environments, and compatibility with flexible fabrication is highly desired for near-space vehicles and polar region applications. Herein, we demonstrate efficient low-temperature flexible perovskite solar cells by improving the interfacial contact between electron-transport layer (ETL) and perovskite layer. We find that the adsorbed oxygen active sites and oxygen vacancies of flexible tin oxide (SnO₂) ETL layer can be effectively decreased by incorporating a trace amount of titanium tetrachloride (TiCl₄). The effective defects elimination at the interfacial increases the electron mobility of flexible SnO₂ layer, regulates band alignment at the perovskite/SnO₂ interface, induces larger perovskite crystal growth, and improves charge collection efficiency in a complete solar cell. Correspondingly, the improved interfacial contact transforms into high-performance solar cells under one-sun illumination (AM 1.5G) with efficiencies up to 23.7 % at 218 K, which might open up a new era of application of this emerging flexible photovoltaic technology to low-temperature environments such as near-space and polar regions.     

Highly Dispersed and Nano-sized Pt-based Electrocatalysts for Low-Temperature Fuel Cells

Among metals, Pt is so far the best material to be used as anode and cathode in low-temperature fuel cells. However, Pt has the drawback of being expensive and easily CO-poisoned. Thus, to produce useful electrocatalysts, significant efforts have been made worldwide on developing Pt-based catalysts with low Pt contents as well as searching for alternative materials with high catalytic activity for anodic and cathodic reactions in low-temperature fuel cells. This article presents the development of highly dispersed and nano-sized Pt-based electrocatalysts synthesized by several new methods based on our experimental results. In the case of anode materials, our proposed new method consists of the synthesis of Pt-based nanoparticles in order to maximize their surface availability, combined with the use of secondary metals that promote the oxidations of methanol and CO. On the other hand, for the cathode materials, the use of the Pt catalysts mixed with metal oxides enhances their oxygen reduction reaction (ORR) activity. We anticipate that the highly dispersed Pt-based nanoparticles introduced in this article will improve the performance of anode and cathode for low-temperature fuel cells.     

Adsorption isotherm for uranyl biosorption by Saccharomyces cerevisiae biomass

Biosorption of uranyl ions from aqueous solution by Saccharomyces cerevisiae was studied in a batch system. The influence of contact time, initial pH, temperature and initial concentration was investigated. The optimal conditions were found to be 3.5?h of contact time and pH?=?4.5. Temperature had no significant effect on adsorption. The uptake of uranyl ions was relatively fast and 85?% of the sorption was completed within 10?min. The experimental data were well fitted with Langmuir isotherm model and pseudo-second order kinetic model. According to this kinetic model, the sorption capacity and the rate constant were 0.455?mmol UO₂ ²⁺/g dry biomass and 1.89?g?mmol^?1?min^?1, respectively. The Langmuir isotherm indicated high affinity and capacity of the adsorbent for uranyl biosorption with the maximum loading of 0.477?mmol UO₂ ²⁺/g dry weight.     

Impact of High Temperature on Ethanol Fermentation by Kluyveromyces marxianus Immobilized on Banana Leaf Sheath Pieces

Ethanol fermentation was carried out with Kluyveromyces marxianus cells at various temperatures (30, 35, 40, and 45 °C). Fermentation performance of the immobilized yeast on banana leaf sheath pieces and the free yeast were evaluated and compared. Generally, ethanol production of the immobilized and free yeast was stable in a temperature range of 30–40 °C. Temperature of 45 °C restricted yeast growth and lengthened the fermentation. The immobilized yeast demonstrated faster sugar assimilation and higher ethanol level in the fermentation broth in comparison with the free yeast at all fermentation temperatures. Change in fatty acid level in cellular membrane was determined to clarify the response of the free and immobilized yeast to thermal stress. The free cells of K. marxianus responded to temperature increase by increasing saturated fatty acid (C16:0 and C18:0) level and by decreasing unsaturated fatty acid (C18:1 and C18:2) level in cellular membrane. For fermentation at 40 °C with immobilized cells of K. marxianus, however, the changes were not observed in both saturated fatty acid (C16:0) and unsaturated fatty acid (C18:1 and C18:2) level.     

Bioaccumulation of thorium and uranyl ions on Saccharomyces cerevisiae

The efficiency of the yeast suspension ofSaccharomyces cerevisiae was studied in the decontamination of radioactive residual waste waters and in the biological behavior of the levuric mass. The effect of reaction time on the amino acid content in the supernatant and on the concentration of radioactive ions was also investigated. The reaction of yeast with the toxic agent showed that the living matter behaves differently from the chemical components of the cell. The yeast had the tendency to aggregate in the presence of radioactive ions. This revised version was published online in July 2006 with corrections to the Cover Date.     

Immobilized Cells in Anaerobic Digestion for Methane from Poultry Manure

The two-stage immobilized microbe waste processor designed for sewage treatment by Messing has been modified to process poultry manure. The Messing reactor of 120-mL volume was modified and scaled up to a 4-L volume. Three different carrier materials have been investigated. Temperatures for each of the two stages were examined, and residence time as well as feed concentration were explored. Analytical data has been computer analyzed using multiple variable correlations and the results of this analysis have indicated directions for optimization.     

The influence of inert solids on ethanol production by Saccharomyces cerevisiae

The catalytic role of various inert solid supports on acceleration of alcohol fermentation by Saccharomyces cerevisiae was investigated. The enhanced rate of alcohol production was dependent on the nature of the support as well as on the amount used. Among all the tested supports, chitosan flakes showed the maximum yield of alcohol (93% of theoretical yield). This higher rate of alcohol production was associated with the twofold increase in the activity of alcohol dehydrogenase over control. Our results suggest that the addition of a small fraction of solids in submerged fermentations to facilitate cell anchorage for enhanced metabolic activity is easier and more economical compared to cell immobilization processes. IICT Communication No. 4266. Some of the results in this article are covered under a patent.     

Photodynamic activities of food additive dyes on the yeast Saccharomyces cerevisiae

Photodynamic cell-inactivating activities of food additive dyes on the yeast Saccharomyces cerevisiae were investigated. Activities of dyes not permitted as food additives were also examined. Red No. 105 (rose bengal), Red No. 3 (erythrosine) and Red No. 104 (phloxine), which are permitted as food additives, markedly inactivated yeast cells by photodynamic action. Eosine, matius yellow and guinia green B, which are not permitted, also exhibited moderate cell-inactivating activity by photodynamic action. None of the dyes used in this experiment exhibited petite induction by photodynamic action.     

Surface Display of Malolactic Enzyme from Oenococcus oeni on Saccharomyces cerevisiae

In order to display malolactic enzyme (MLE) on the cell surface of Saccharomyces cerevisiae, a yeast cell surface display plasmid pADH1-AGG was constructed by fusing the α-factor signal encoding sequence (267 bp) and the C-terminal half of α-agglutinin encoding sequence (1,645 bp) into the plasmid pADH1. The pADH1-AGG could successfully express and anchor the enhanced green fluorescent protein (EGFP) onto the yeast cell surface when the EGFP was used to verify its function. Then the pADH1-MLE was constructed by inserting the MLE encoding sequence (1,600 bp) into the pADH1-AGG and introduced into S. cerevisiae cells. The positive strain carrying pADH1-MLE was confirmed by use of the 6× His monoclonal antibody and fluorescein isothiocyanate-conjugated goat anti-mouse IgG. All results indicated that the MLE was displayed successfully on the cell surface of positive transformant. The MLE activity of genetically engineered yeast strain could turn 21.11 % l-malate into lactic acid after 12 h reaction with l-malate. The constructed yeast strain might be used to conduct malolactic fermentation (MLF) in wine to solve the important issues of sluggish MLF, microbial spoilage, and adverse metabolic substances produced by the lactic acid bacteria.     

A proteomic approach for the study of Saccharomyces cerevisiae cell wall biogenesis

In fungi, cell shape is determined by the presence of a rigid cell wall which separates the cell from the extracellular medium. This highly dynamic structure is essential for the maintenance of cell integrity and is involved in several phenomena such as flocculation, adherence and pathogenicity. The composition of the fungal cell wall is well known, but issues such as the assembly and remodeling of its components remain poorly understood. In an attempt to study the de novo construction of the yeast cell wall, we have undertaken a large-scale proteomic approach to analyze the proteins secreted by regenerating protoplasts. Upon incubation of protoplasts in regenerating conditions, numerous proteins are secreted into the culture medium. These presumably include proteins destined for the cell wall, comprising both structural proteins as well as enzymes involved in cell wall biogenesis. This work reports the establishment of a reference map of proteins secreted by regenerating protoplasts by means of two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and their identification by mass spectrometry. Thirty-two different proteins have been identified, including known cell wall proteins, glycolytic enzymes, heat shock proteins, and proteins involved in several other processes. Using this approach, novel proteins possibly involved in cell wall construction have also been identified. This reference map will allow comparative analyses to be carried out on a selected collection of mutants affected in the cell wall.     

Effect of agitation and aeration on production of hexokinase by Saccharomyces cerevisiae

A batch culture of Saccharomyces cerevisiae for the production of hexokinase was carried out in a 5-L fermentor containing 3 L of culture medium, which was in oculated with cell suspension (about 0.7 g/L), and left ferm entingat 35°C and pH 4.0. The aeration and agitation were adjusted to attain k _La values of 15, 60, 135, and 230 h⁻¹. The highest hexokinase productivity (754.6 U/[L h]) and substrate-cell conversion yield (0.21 g/g) occurred for a k _La of 60 h⁻¹. Moreover, the formation of hexokinase and cell growth are coupled events, which is in accordance with the constitutive character of this enzyme. Hexokinase formation for k _La>60 h⁻¹ was not enhanced probably owing to saturation of the respiratory pathway by oxygen.     

Effects of low-frequency magnetic fields on the viability of yeast Saccharomyces cerevisiae

A 50 Hz magnetic field effect on the growth of yeasts Saccharomyces cerevisae was studied. The cylindrical coil induced magnetic fields with inductions up to 10 mT. Duration of exposure varied up to 24 min. Exposure took place at laboratory temperature (24-26 degrees C) and the air ventilator maintained the temperature at the place of the sample. We measured the growth curves of yeasts in broth and we calculated the number of CFU (colony forming units) on solid soil. We found that magnetic field decreases the number of yeasts, and slowed down their growth. The result is similar to the experiments with bacteria E. coli, S. aureus and L. adecarboxylata. It seems that the magnetic fields kill a part of yeasts and the bigger part of them survives and continues in their growth.     

Physicochemical property based computational scheme for classifying DNA sequence elements of Saccharomyces cerevisiae

GenerationE of huge “omics” data necessitates the development and application of computational methods to annotate the data in terms of biological features. In the context of DNA sequence, it is important to unravel the hidden physicochemical signatures. For this purpose, we have considered various sequence elements such as promoter, ACS, LTRs, telomere, and retrotransposon of the model organism Saccharomyces cerevisiae. Contributions due to di-nucleotides play a major role in studying the DNA conformation profile. The physicochemical parameters used are hydrogen bonding energy, stacking energy and solvation energy per base pair. Our computational study shows that all sequence elements in this study have distinctive physicochemical signatures and the same can be exploited for prediction experiments. The order that we see in a DNA sequence is dictated by biological regions and hence, there exists role of dependency in the sequence makeup, keeping this in mind we are proposing two computational schemes (a) using a windowing block size procedure and (b) using di-nucleotide transitions. We obtained better discriminating profile when we analyzed the sequence data in windowing manner. In the second novel approach, we introduced the di-nucleotide transition probability matrix (DTPM) to study the hidden layer of information embedded in the sequences. DTPM has been used as weights for scanning and predictions. This proposed computational scheme incorporates the memory property which is more realistic to study the physicochemical properties embedded in DNA sequences. Our analysis shows that the DTPM scheme performs better than the existing method in this applied region. Characterization of these elements will be a key to genome editing applications and advanced machine learning approaches may also require such distinctive profiles as useful input features.     

培养基碳源对固定化酵母细胞催化乙酰基三甲基硅烷不对称还原反应的影响

 尽管含芳香基等较大取代基团的底物能被生物转化为高对映体纯手性有机硅醇,但生物催化其他底物还原反应的产物收率及对映体选择性均较低. 作者筛选到一株能高效催化羰基/羟基不对称转化的酵母菌株,探讨了用固定化酵母细胞高效催化乙酰基三甲基硅烷不对称还原制1-三甲基硅乙醇的可能性,系统研究了培养基碳源及其浓度对该反应的影响. 发现在水/正己烷两相体系中,固定化酵母细胞能催化乙酰基三甲基硅烷不对称还原成1-三甲基硅乙醇. 可通过调节培养基碳源的种类及其浓度在一定程度上控制反应的产物收率及对映体选择性. 以最适碳源(3.0%麦芽汁)培养的酵母细胞催化该反应的产物收率和对映体过量值分别为91.3%和72.8%,远高于文献报道值.     

Mediated amperometric biosensors for lactic acid based on carbon paste electrodes modified with baker's yeast Saccharomyces cerevisiae

Carbon paste electrodes modified with baker's yeast Saccharomyces cerevisiae (a source of flavocytochrome b(2)) were investigated as amperometric biosensors for lactic acid. Phenazine methosulphate was used as a mediator. The optimal operational conditions of the electrodes were: an operating potential 0.0 V, solution pH 7.2, concentration of phenazine methosulphate in solution 0.2 mM. A linear range in the dependence of the current responses on the concentration of lactic acid was up to 1 mM. The suitability of the electrodes for determination of lactic acid in milk and dairy products such as kefir and yoghurt was tested. The yeast cells in the paste remained viable at least for 1 month.     

Adaptive Evolution of Saccharomyces cerevisiae with Enhanced Ethanol Tolerance for Chinese Rice Wine Fermentation

High tolerance towards ethanol is a desirable property for the Saccharomyces cerevisiae strains used in the alcoholic beverage industry. To improve the ethanol tolerance of an industrial Chinese rice wine yeast, a sequential batch fermentation strategy was used to adaptively evolve a chemically mutagenized Chinese rice wine G85 strain. The high level of ethanol produced under Chinese rice wine-like fermentation conditions was used as the selective pressure. After adaptive evolution of approximately 200 generations, mutant G85X-8 was isolated and shown to have markedly increased ethanol tolerance. The evolved strain also showed higher osmotic and temperature tolerances than the parental strain. Laboratory Chinese rice wine fermentation showed that the evolved G85X-8 strain was able to catabolize sugars more completely than the parental G85 strain. A higher level of yeast cell activity was found in the fermentation mash produced by the evolved strain, but the aroma profiles were similar between the evolved and parental strains. The improved ethanol tolerance in the evolved strain might be ascribed to the altered fatty acids composition of the cell membrane and higher intracellular trehalose concentrations. These results suggest that adaptive evolution is an efficient approach for the non-recombinant modification of industrial yeast strains.