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Sample records for cerevisiae microbial cell

  1. Microbial Cells as Biosorbents for Heavy Metals: Accumulation of Uranium by Saccharomyces cerevisiae and Pseudomonas aeruginosa

    PubMed Central

    Strandberg, Gerald W.; Shumate, Starling E.; Parrott, John R.

    1981-01-01

    Uranium accumulated extracellularly on the surfaces of Saccharomyces cerevisiae cells. The rate and extent of accumulation were subject to environmental parameters, such as pH, temperature, and interference by certain anions and cations. Uranium accumulation by Pseudomonas aeruginosa occurred intracellularly and was extremely rapid (<10 s), and no response to environmental parameters could be detected. Metabolism was not required for metal uptake by either organism. Cell-bound uranium reached a concentration of 10 to 15% of the dry cell weight, but only 32% of the S. cerevisiae cells and 44% of the P. aeruginosa cells within a given population possessed visible uranium deposits when examined by electron microscopy. Rates of uranium uptake by S. cerevisiae were increased by chemical pretreatment of the cells. Uranium could be removed chemically from S. cerevisiae cells, and the cells could then be reused as a biosorbent. Images PMID:16345691

  2. Sensitive determination of L-lysine with a new amperometric microbial biosensor based on Saccharomyces cerevisiae yeast cells.

    PubMed

    Akyilmaz, Erol; Erdoğan, Ali; Oztürk, Ramazan; Yaşa, Ihsan

    2007-01-15

    A new amperometric microbial biosensor based on Saccharomyces cerevisiae NRRL-12632 cells, which had been induced for lysine oxidase enzyme and immobilized in gelatin by a cross-linking agent was developed for the sensitive determination of L-lysine amino acid. To construct the microbial biosensor S. cerevisiae cells were activated and cultured in a suitable culture medium. By using gelatine (8.43 mg cm(-2)) and glutaraldehyde (0.25%), cells obtained in the logarithmic phase of the growth curve at the end of a 14 h period were immobilized and fixed on a pretreated oxygen sensitive Teflon membrane of a dissolved oxygen probe. The assay procedure of the microbial biosensor is based on the determination of the differences of the respiration activity of the cells on the oxygenmeter in the absence and the presence of L-lysine. According to the end point measurement technique used in the experiments it was determined that the microbial biosensor response depended linearly on L-lysine concentrations between 1.0 and 10.0 microM with a 1 min response time. In optimization studies of the microbial biosensor, the most suitable microorganism quantities were found to be 0.97x10(5)CFU cm(-2). In addition phosphate buffer (pH 7.5; 50 mM) and 30 degrees C were obtained as the optimum working conditions. In characterization studies of the microbial biosensor some parameters such as substrate specificity, interference effects of some substances on the microbial biosensor responses, reproducibility of the biosensor and operational and storage stability were investigated. PMID:16759846

  3. Dominance of Saccharomyces cerevisiae in alcoholic fermentation processes: role of physiological fitness and microbial interactions.

    PubMed

    Albergaria, Helena; Arneborg, Nils

    2016-03-01

    Winemaking, brewing and baking are some of the oldest biotechnological processes. In all of them, alcoholic fermentation is the main biotransformation and Saccharomyces cerevisiae the primary microorganism. Although a wide variety of microbial species may participate in alcoholic fermentation and contribute to the sensory properties of end-products, the yeast S. cerevisiae invariably dominates the final stages of fermentation. The ability of S. cerevisiae to outcompete other microbial species during alcoholic fermentation processes, such as winemaking, has traditionally been ascribed to its high fermentative power and capacity to withstand the harsh environmental conditions, i.e. high levels of ethanol and organic acids, low pH values, scarce oxygen availability and depletion of certain nutrients. However, in recent years, several studies have raised evidence that S. cerevisiae, beyond its remarkable fitness for alcoholic fermentation, also uses defensive strategies mediated by different mechanisms, such as cell-to-cell contact and secretion of antimicrobial peptides, to combat other microorganisms. In this paper, we review the main physiological features underlying the special aptitude of S. cerevisiae for alcoholic fermentation and discuss the role of microbial interactions in its dominance during alcoholic fermentation, as well as its relevance for winemaking. PMID:26728020

  4. Microbial Cell Imaging

    SciTech Connect

    Doktycz, Mitchel John; Sullivan, Claretta; Mortensen, Ninell P; Allison, David P

    2011-01-01

    the maximum scan size (roughly 100 x 100 {mu}m) and the restricted movement of the cantilever in the Z (or height) direction. In most commercial AFMs, the Z range is restricted to roughly 10 {mu}m such that the height of cells to be imaged must be seriously considered. Nevertheless, AFM can provide structural-functional information at nanometer resolution and do so in physiologically relevant environments. Further, instrumentation for scanning probe microscopy continues to advance. Systems for high-speed imaging are becoming available, and techniques for looking inside the cells are being demonstrated. The ability to combine AFM with other imaging modalities is likely to have an even greater impact on microbiological studies. AFM studies of intact microbial cells started to appear in the literature in the 1990s. For example, AFM studies of Saccharomyces cerevisiae examined buddings cars after cell division and detailed changes related to cell growth processes. Also, the first AFM studies of bacterial biofilms appeared. In the late 1990s, AFM studies of intact fungal spores described clear changes in spore surfaces upon germination, and studies of individual bacterial cells were also described. These early bacterial imaging studies examined changes in bacterial morphology due to antimicrobial peptides exposure and bacterial adhesion properties. The majority of these early studies were carried out on dried samples and took advantage of the resolving power of AFM. The lack of cell mounting procedures presented an impediment for cell imaging studies. Subsequently, several approaches to mounting microbial cells have been developed, and these techniques are described later. Also highlighted are general considerations for microbial imaging and a description of some of the various applications of AFM to microbiology.

  5. Metabolic Differences in Microbial Cell Populations Revealed by Nanophotonic Ionization

    SciTech Connect

    Walker, Bennett; Antonakos, Cory; Retterer, Scott T; Vertes, Akos

    2013-01-01

    ellular differences are linked to cell differentiation, the proliferation of cancer and to the development of drug resistance in microbial infections. Due to sensitivity limitations, however, large- scale metabolic analysis at the single cell level is only available for cells significantly larger in volume than Saccharomyces cerevisiae (~30 fL). Here we demonstrate that by a nanophotonic ionization platform and mass spectrometry, over one hundred up to 108 metabolites, or up to 18% of the known S. cerevisiae metabolome, can be identified in very small cell populations (n < 100). Under ideal conditions, r Relative quantitation of up to 4% of the metabolites is achieved at the single cell level.

  6. Exploration and comparison of inborn capacity of aerobic and anaerobic metabolisms of Saccharomyces cerevisiae for microbial electrical current production

    PubMed Central

    Mao, Longfei; Verwoerd, Wynand S

    2013-01-01

    Saccharomyces cerevisiae possesses numerous advantageous biological features, such as being robust, easily handled, mostly non-pathogenic and having high catabolic rates, etc., which can be considered as merits for being used as a promising biocatalyst in microbial fuel cells (MFCs) for electricity generation. Previous studies have developed efficient MFC configurations to convert metabolic electron shuttles, such as cytoplasmic NADH, into usable electric current. However, no studies have elucidated the maximum potential of S. cerevisiae for current output and the underlying metabolic pathways, resulting from the interaction of thousands of reactions inside the cell during MFC operation. To address these two key issues, this study used in silico metabolic engineering techniques, flux balance analysis (FBA), and flux variability analysis with target flux minimization (FATMIN), to model the metabolic perturbation of S. cerevisiae under the MFC-energy extraction. The FBA results showed that, in the cytoplasmic NADH-dependent mediated electron transfer (MET) mode, S. cerevisiae had a potential to produce currents at up to 5.781 A/gDW for the anaerobic and 6.193 A/gDW for the aerobic environments. The FATMIN results showed that the aerobic and anaerobic metabolisms are resilient, relying on six and five contributing reactions respectively for high current production. Two reactions, catalyzed by glutamate dehydrogenase (NAD) (EC 1.4.1.3) and methylene tetrahydrofolate dehydrogenase (NAD) (EC 1.5.1.5), were shared in both current-production modes and contributed to over 80% of the identified maximum current outputs. It is also shown that the NADH regeneration was much less energy costly than biomass production rate. Taken together, our finding suggests that S. cerevisiae should receive more research effort for MFC electricity production. PMID:23969939

  7. Effects of spaceflight on polysaccharides of Saccharomyces cerevisiae cell wall.

    PubMed

    Liu, Hong-Zhi; Wang, Qiang; Liu, Xiao-Yong; Tan, Sze-Sze

    2008-12-01

    Freeze-dried samples of four Saccharomyces cerevisiae strains, namely, FL01, FL03, 2.0016, and 2.1424, were subjected to spaceflight. After the satellite's landing on Earth, the samples were recovered and changes in yeast cell wall were analyzed. Spaceflight strains of all S. cerevisiae strains showed significant changes in cell wall thickness (P < 0.05). One mutant of S. cerevisiae 2.0016 with increased biomass, cell wall thickness, and cell wall glucan was isolated (P < 0.05). The spaceflight mutant of S. cerevisiae 2.0016 showed 46.7%, 62.6%, and 146.0% increment in biomass, cell wall thickness and beta-glucan content, respectively, when compared to the ground strain. Moreover, growth curve analysis showed spaceflight S. cerevisiae 2.0016 had a faster growth rate, shorter lag phase periods, higher final biomass, and higher content of beta-glucan. Genetic stability analysis showed that prolonged subculturing of spaceflight strain S. cerevisiae 2.0016 did not lead to the appearance of variants, indicating that the genetic stability of S. cerevisiae 2.0016 mutant could be sufficient for its exploitation of beta-glucan production. PMID:18797865

  8. Glucose-based microbial production of the hormone melatonin in yeast Saccharomyces cerevisiae.

    PubMed

    Germann, Susanne M; Baallal Jacobsen, Simo A; Schneider, Konstantin; Harrison, Scott J; Jensen, Niels B; Chen, Xiao; Stahlhut, Steen G; Borodina, Irina; Luo, Hao; Zhu, Jiangfeng; Maury, Jérôme; Forster, Jochen

    2016-05-01

    Melatonin is a natural mammalian hormone that plays an important role in regulating the circadian cycle in humans. It is a clinically effective drug exhibiting positive effects as a sleep aid and a powerful antioxidant used as a dietary supplement. Commercial melatonin production is predominantly performed by complex chemical synthesis. In this study, we demonstrate microbial production of melatonin and related compounds, such as serotonin and N-acetylserotonin. We generated Saccharomyces cerevisiae strains that comprise heterologous genes encoding one or more variants of an L-tryptophan hydroxylase, a 5-hydroxy-L-tryptophan decarboxylase, a serotonin acetyltransferase, an acetylserotonin O-methyltransferase, and means for providing the cofactor tetrahydrobiopterin via heterologous biosynthesis and recycling pathways. We thereby achieved de novo melatonin biosynthesis from glucose. We furthermore accomplished increased product titers by altering expression levels of selected pathway enzymes and boosting co-factor supply. The final yeast strain produced melatonin at a titer of 14.50 ± 0.57 mg L(-1) in a 76h fermentation using simulated fed-batch medium with glucose as sole carbon source. Our study lays the basis for further developing a yeast cell factory for biological production of melatonin. PMID:26710256

  9. Applications of Microbial Cell Sensors

    NASA Astrophysics Data System (ADS)

    Shimomura-Shimizu, Mifumi; Karube, Isao

    Since the first microbial cell sensor was studied by Karube et al. in 1977, many types of microbial cell sensors have been developed as analytical tools. The microbial cell sensor utilizes microbes as a sensing element and a transducer. The characteristics of microbial cell sensors as sensing devices are a complete contrast to those of enzyme sensors or immunosensors, which are highly specific for the substrates of interest, although the specificity of the microbial cell sensor has been improved by genetic modification of the microbe used as the sensing element. Microbial cell sensors have the advantages of tolerance to measuring conditions, a long lifetime, and good cost performance, and have the disadvantage of a long response time. In this review, applications of microbial cell sensors are summarized.

  10. Microbial sensor cell arrays.

    PubMed

    Melamed, Sahar; Elad, Tal; Belkin, Shimshon

    2012-02-01

    Motivated by the advantages endowed by high-throughput analysis, researchers have succeeded in incorporating multiple reporter cells into a single platform; the technology now allows the simultaneous scrutiny of a large collection of sensor strains. We review current aspects in cell array technology with emphasis on microbial sensor arrays. We consider various techniques for patterning live cells on solid surfaces, describe different array-based applications and devices, and highlight recent efforts for live cell storage. We review mathematical approaches for deciphering the data emanating from bioreporter collections, and discuss the future of single cell arrays. Innovative technologies for cell patterning, preservation and interpretation are continuously being developed; when they all mature, cell arrays may become an efficient analytical tool, in a scope resembling that of DNA microarray biochips. PMID:22176747

  11. Metabolic engineering of Saccharomyces cerevisiae: a key cell factory platform for future biorefineries.

    PubMed

    Hong, Kuk-Ki; Nielsen, Jens

    2012-08-01

    Metabolic engineering is the enabling science of development of efficient cell factories for the production of fuels, chemicals, pharmaceuticals, and food ingredients through microbial fermentations. The yeast Saccharomyces cerevisiae is a key cell factory already used for the production of a wide range of industrial products, and here we review ongoing work, particularly in industry, on using this organism for the production of butanol, which can be used as biofuel, and isoprenoids, which can find a wide range of applications including as pharmaceuticals and as biodiesel. We also look into how engineering of yeast can lead to improved uptake of sugars that are present in biomass hydrolyzates, and hereby allow for utilization of biomass as feedstock in the production of fuels and chemicals employing S. cerevisiae. Finally, we discuss the perspectives of how technologies from systems biology and synthetic biology can be used to advance metabolic engineering of yeast. PMID:22388689

  12. Microbial fuel cells

    SciTech Connect

    Nealson, Kenneth H; Pirbazari, Massoud; Hsu, Lewis

    2013-04-09

    A microbial fuel cell includes an anode compartment with an anode and an anode biocatalyst and a cathode compartment with a cathode and a cathode biocatalyst, with a membrane positioned between the anode compartment and the cathode compartment, and an electrical pathway between the anode and the cathode. The anode biocatalyst is capable of catalyzing oxidation of an organic substance, and the cathode biocatalyst is capable of catalyzing reduction of an inorganic substance. The reduced organic substance can form a precipitate, thereby removing the inorganic substance from solution. In some cases, the anode biocatalyst is capable of catalyzing oxidation of an inorganic substance, and the cathode biocatalyst is capable of catalyzing reduction of an organic or inorganic substance.

  13. Microbial Fuel Cells and Microbial Electrolyzers

    SciTech Connect

    Borole, Abhijeet P

    2015-01-01

    Microbial Fuel Cells and microbial electrolyzers represent an upcoming technology for production of electricity and hydrogen using a hybrid electrocatalytic-biocatalytic approach. The combined catalytic efficiency of these processes has potential to make this technology highly efficient among the various renewable energy production alternatives. This field has attracted electrochemists, biologists and many other disciplines due to its potential to contribute to the energy, water and environment sectors. A brief introduction to the technology is provided followed by current research needs from a bioelectrochemical perspective. Insights into the operation and limitations of these systems achieved via cyclic voltammetry and impedance spectroscopy are discussed along with the power management needs to develop the application aspects. Besides energy production, other potential applications in bioenergy, bioelectronics, chemical production and remediation are also highlighted.

  14. Evaluation of microbial qPCR workflows using engineered Saccharomyces cerevisiae

    PubMed Central

    Da Silva, S.M.; Vang, L.K.; Olson, N.D.; Lund, S.P.; Downey, A.S.; Kelman, Z.; Salit, M.L.; Lin, N.J.; Morrow, J.B.

    2016-01-01

    Aims We describe the development and interlaboratory study of modified Saccharomyces cerevisiae as a candidate material to evaluate a full detection workflow including DNA extraction and quantitative polymerase chain reaction (qPCR). Methods and results S. cerevisiae NE095 was prepared by stable insertion of DNA sequence External RNA Control Consortium-00095 into S. cerevisiae BY4739 to convey selectivity. For the interlaboratory study, a binomial regression model was used to select three cell concentrations, high (4 × 107 cells ml−1), intermediate (4 × 105 cells ml−1) and low (4 × 103 cells ml−1), and the number of samples per concentration. Seven participants, including potential end users, had combined rates of positive qPCR detection (quantification cycle <37) of 100%, 40%, and 0% for high, intermediate, and low concentrations, respectively. Conclusions The NE095 strain was successfully detected by all participants, with the high concentration indicating a potential target concentration for a reference material. Significance and impact of the study The engineered yeast has potential to support measurement assurance for the analytical process of qPCR, encompassing the method, equipment, and operator, to increase confidence in results and better inform decision-making in areas of applied microbiology. This material can also support process assessment for other DNA-based detection technologies. PMID:27077050

  15. Micromachined microbial and photosynthetic fuel cells

    NASA Astrophysics Data System (ADS)

    Chiao, Mu; Lam, Kien B.; Lin, Liwei

    2006-12-01

    This paper presents two types of fuel cells: a miniature microbial fuel cell (µMFC) and a miniature photosynthetic electrochemical cell (µPEC). A bulk micromachining process is used to fabricate the fuel cells, and the prototype has an active proton exchange membrane area of 1 cm2. Two different micro-organisms are used as biocatalysts in the anode: (1) Saccharomyces cerevisiae (baker's yeast) is used to catalyze glucose and (2) Phylum Cyanophyta (blue-green algae) is used to produce electrons by a photosynthetic reaction under light. In the dark, the µPEC continues to generate power using the glucose produced under light. In the cathode, potassium ferricyanide is used to accept electrons and electric power is produced by the overall redox reactions. The bio-electrical responses of µMFCs and µPECs are characterized with the open-circuit potential measured at an average value of 300-500 mV. Under a 10 ohm load, the power density is measured as 2.3 nW cm-2 and 0.04 nW cm-2 for µMFCs and µPECs, respectively.

  16. Directed Evolution of Metabolic Pathways in Microbial Populations. I. Modification of the Acid Phosphatase Ph Optimum in S. CEREVISIAE

    PubMed Central

    Francis, J. C.; Hansche, P. E.

    1972-01-01

    An experimental system for directing the evolution of enzymes and metabolic pathways in microbial populations is proposed and an initial test of its power is provided.—The test involved an attempt to genetically enhance certain functional properties of the enzyme acid phosphatase in S. cerevisiae by constructing an environment in which the functional changes desired would be "adaptive". Naturally occurring mutations in a population of 109 cells were automatically and continuously screened, over 1,000 generations, for their effect on the efficiency (Km) and activity of acid phosphatase at pH 6, and for their effect on the efficiency of orthophosphate metabolism.—The first adaptation observed, M1, was due to a single mutational event that effected a 30% increase in the efficiency of orthophosphate metabolism. The second, M2, effected an adaptive shift in the pH optimum of acid phosphatase and an increase in its activity over a wide range of pH values (an increment of 60% at pH 6). M2 was shown to result from a single mutational event in the region of the acid phosphatase structural gene. The third, M3, effected cell clumping, an adaptation to the culture apparatus that had no effect on phosphate metabolism.—The power of this system for directing the evolution of enzymes and of metabolic pathways is discussed in terms of the kinetic properties of the experimental system and in terms of the results obtained. PMID:4552227

  17. Effects of Saccharomyces cerevisiae fermentation products on dairy calves: Ruminal fermentation, gastrointestinal morphology, and microbial community.

    PubMed

    Xiao, J X; Alugongo, G M; Chung, R; Dong, S Z; Li, S L; Yoon, I; Wu, Z H; Cao, Z J

    2016-07-01

    The aim of this study was to evaluate the effects of Saccharomyces cerevisiae fermentation products (SCFP) in the calf starter and milk on ruminal fermentation, gastrointestinal morphology, and microbial community in the first 56 d of life. Thirty Holstein bull calves were randomly assigned to 1 of 3 groups: a texturized calf starter containing 0 (CON), 0.5, or 1% SCFP (XPC, Diamond V, Cedar Rapids, IA) of dry matter from d 4 to 56. In addition, the XPC-supplemented calves were fed with 1 g/d SCFP (SmartCare, Diamond V, Cedar Rapids, IA) in milk from d 2 to 30. All calves were fed 4 L of colostrum within 1 h of birth and were subsequently fed milk twice daily until weaned on d 56. Rumen fluid was collected by an esophageal tube 4 h after the morning feeding on d 28 and 56 to determine ruminal pH, ammonia-N, and volatile fatty acids concentrations. On d 56, 15 (5 per treatment) calves were harvested and slaughter weight, gastrointestinal morphology parameters, and bacteria community were recorded. Papilla length, width, and surface area were measured from 5 locations within the rumen. Villus height, width, surface area, crypt depth, and villus height-to-crypt depth ratio were measured in the duodenum, jejunum, and ileum. Next-generation sequencing technology was used to test the microbial community of the rumen and duodenum samples on d 28 and 56. Data were analyzed by MIXED procedure in SAS (SAS Institute Inc., Cary, NC) with contrast statements to declare CON versus all SCFP and 0.5 versus 1% SCFP in starter grains. Ruminal pH, ammonia-N, and total volatile fatty acids were not altered by SCFP. However, the supplemented groups exhibited higher ruminal butyrate concentrations coinciding with higher Butyrivibrio and lower Prevotella richness than CON group. Supplementation of SCFP increased papilla length in the rumen. In the small intestine, SCFP reduced crypt depth of jejunum, and increased villus height-to-crypt depth ratio in all segments of the small intestine

  18. Catalytic activity of baker's yeast in a mediatorless microbial fuel cell.

    PubMed

    Sayed, Enas Taha; Tsujiguchi, Takuya; Nakagawa, Nobuyoshi

    2012-08-01

    The catalytic activity of baker's yeast, Saccharomyces cerevisiae, as a biocatalyst was investigated in a mediatorless microbial fuel cell. The yeast cells that adhered on the anode surface were the active biocatalyst for glucose oxidation in a mediatorless biofuel cell, suggesting that the electron transfer took place through the surface confined species. The species in the anolyte solution including the dispersed yeast cells did not take a part in the electron transfer and thus in the power generation. PMID:22357359

  19. Symmetric cell division in pseudohyphae of the yeast Saccharomyces cerevisiae.

    PubMed Central

    Kron, S J; Styles, C A; Fink, G R

    1994-01-01

    Laboratory strains of Saccharomyces cerevisiae are dimorphic; in response to nitrogen starvation they switch from a yeast form (YF) to a filamentous pseudohyphal (PH) form. Time-lapse video microscopy of dividing cells reveals that YF and PH cells differ in their cell cycles and budding polarity. The YF cell cycle is controlled at the G1/S transition by the cell-size checkpoint Start. YF cells divide asymmetrically, producing small daughters from full-sized mothers. As a result, mothers and daughters bud asynchronously. Mothers bud immediately but daughters grow in G1 until they achieve a critical cell size. By contrast, PH cells divide symmetrically, restricting mitosis until the bud grows to the size of the mother. Thus, mother and daughter bud synchronously in the next cycle, without a G1 delay before Start. YF and PH cells also exhibit distinct bud-site selection patterns. YF cells are bipolar, producing their second and subsequent buds at either pole. PH cells are unipolar, producing their second and subsequent buds only from the end opposite the junction with their mother. We propose that in PH cells a G2 cell-size checkpoint delays mitosis until bud size reaches that of the mother cell. We conclude that yeast and PH forms are distinct cell types each with a unique cell cycle, budding pattern, and cell shape. Images PMID:7841518

  20. Immobilized cell cross-flow reactor. [Saccharomyces cerevisiae

    SciTech Connect

    Chotani, G.K.; Constantinides, A.

    1984-01-01

    A cross-current flow reactor was operated using sodium alginate gel entrapped yeast cells (Saccharomyces cerevisiae) under growth conditions. Micron-sized silica, incorporated into the biocatalyst particles (1 mm mean diameter) improved mechanical strength and internal surface adhesion. The process showed decreased productivity and stability at 35/sup 0/C compared to the normal study done at 30/sup 0/C. The increased number of cross flows diminish the product inhibition effect. The residence time distribution shows that the cross-flow bioreactor system can be approximated to either a train of backmixed fermentors in series or a plug flow fermentor with moderate axial dispersion.

  1. Directed Evolution of Metabolic Pathways in Microbial Populations II. a Repeatable Adaptation in SACCHAROMYCES CEREVISIAE

    PubMed Central

    Francis, J. C.; Hansche, P. E.

    1973-01-01

    A selection experiment was conducted for approximately 1,000 generations in a chemostat population of 109 cells of the haploid yeast, S. cerevisiae. The experiment was designed to enhance genetically the rate at which the external enzyme acid phosphatase catalyzed the hydrolysis of very low concentrations of β-glycerophosphate at an unfavorably high pH. The observed genetic adaptation in this experiment consisted of a mutation (ACP-2) in the acid phosphatase structural gene which effected a shift in the pH optimum of the enzyme and incremented its activity. The effects of ACP-2 and a similar mutation, ACP-1, on acid phosphatase substrate specificity are also reported. PMID:17248616

  2. Microbial transformations of ferulic acid by Saccharomyces cerevisiae and Pseudomonas fluorescens.

    PubMed Central

    Huang, Z; Dostal, L; Rosazza, J P

    1993-01-01

    Saccharomyces cerevisiae (dry baker's yeast) and Pseudomonas fluorescens were used to convert trans-ferulic acid into 4-hydroxy-3-methoxystyrene in 96 and 89% yields, respectively. The metabolites were isolated by solid-phase extraction and analyzed by thin-layer chromatography and high-performance liquid chromatography. The identities of the metabolites were determined by 1H- and 13C-nuclear magnetic resonance spectroscopy and by mass spectrometry. The mechanism of the decarboxylation of ferulic acid was investigated by measuring the degree and position of deuterium incorporated into the styrene derivative from D2O by mass spectrometry and by both proton and deuterium nuclear magnetic resonance spectroscopies. Resting cells of baker's yeast reduced ferulic acid to 4-hydroxy-3-methoxyphenylpropionic acid in 54% yield when incubations were under an argon atmosphere. PMID:8395165

  3. Local Nanomechanical Motion of the Cell Wall of Saccharomyces cerevisiae

    NASA Astrophysics Data System (ADS)

    Pelling, Andrew E.; Sehati, Sadaf; Gralla, Edith B.; Valentine, Joan S.; Gimzewski, James K.

    2004-08-01

    We demonstrate that the cell wall of living Saccharomyces cerevisiae (baker's yeast) exhibits local temperature-dependent nanomechanical motion at characteristic frequencies. The periodic motions in the range of 0.8 to 1.6 kHz with amplitudes of ~3 nm were measured using the cantilever of an atomic force microscope (AFM). Exposure of the cells to a metabolic inhibitor causes the periodic motion to cease. From the strong frequency dependence on temperature, we derive an activation energy of 58 kJ/mol, which is consistent with the cell's metabolism involving molecular motors such as kinesin, dynein, and myosin. The magnitude of the forces observed (~10 nN) suggests concerted nanomechanical activity is operative in the cell.

  4. Towards a microbial thermoelectric cell.

    PubMed

    Rodríguez-Barreiro, Raúl; Abendroth, Christian; Vilanova, Cristina; Moya, Andrés; Porcar, Manuel

    2013-01-01

    Microbial growth is an exothermic process. Biotechnological industries produce large amounts of heat, usually considered an undesirable by-product. In this work, we report the construction and characterization of the first microbial thermoelectric cell (MTC), in which the metabolic heat produced by a thermally insulated microbial culture is partially converted into electricity through a thermoelectric device optimized for low ΔT values. A temperature of 41°C and net electric voltage of around 250-600 mV was achieved with 1.7 L baker's yeast culture. This is the first time microbial metabolic energy has been converted into electricity with an ad hoc thermoelectric device. These results might contribute towards developing a novel strategy to harvest excess heat in the biotechnology industry, in processes such as ethanol fermentation, auto thermal aerobic digestion (ATAD) or bioremediation, which could be coupled with MTCs in a single unit to produce electricity as a valuable by-product of the primary biotechnological product. Additionally, we propose that small portable MTCs could be conceived and inoculated with suitable thermophilic of hyperthermophilic starter cultures and used for powering small electric devices. PMID:23468862

  5. Towards a Microbial Thermoelectric Cell

    PubMed Central

    Rodríguez-Barreiro, Raúl; Abendroth, Christian; Vilanova, Cristina; Moya, Andrés; Porcar, Manuel

    2013-01-01

    Microbial growth is an exothermic process. Biotechnological industries produce large amounts of heat, usually considered an undesirable by-product. In this work, we report the construction and characterization of the first microbial thermoelectric cell (MTC), in which the metabolic heat produced by a thermally insulated microbial culture is partially converted into electricity through a thermoelectric device optimized for low ΔT values. A temperature of 41°C and net electric voltage of around 250–600 mV was achieved with 1.7 L baker’s yeast culture. This is the first time microbial metabolic energy has been converted into electricity with an ad hoc thermoelectric device. These results might contribute towards developing a novel strategy to harvest excess heat in the biotechnology industry, in processes such as ethanol fermentation, auto thermal aerobic digestion (ATAD) or bioremediation, which could be coupled with MTCs in a single unit to produce electricity as a valuable by-product of the primary biotechnological product. Additionally, we propose that small portable MTCs could be conceived and inoculated with suitable thermophilic of hyperthermophilic starter cultures and used for powering small electric devices. PMID:23468862

  6. CRISPR-Cas9 Genome Engineering in Saccharomyces cerevisiae Cells.

    PubMed

    Ryan, Owen W; Poddar, Snigdha; Cate, Jamie H D

    2016-01-01

    This protocol describes a method for CRISPR-Cas9-mediated genome editing that results in scarless and marker-free integrations of DNA into Saccharomyces cerevisiae genomes. DNA integration results from cotransforming (1) a single plasmid (pCAS) that coexpresses the Cas9 endonuclease and a uniquely engineered single guide RNA (sgRNA) expression cassette and (2) a linear DNA molecule that is used to repair the chromosomal DNA damage by homology-directed repair. For target specificity, the pCAS plasmid requires only a single cloning modification: replacing the 20-bp guide RNA sequence within the sgRNA cassette. This CRISPR-Cas9 protocol includes methods for (1) cloning the unique target sequence into pCAS, (2) assembly of the double-stranded DNA repair oligonucleotides, and (3) cotransformation of pCAS and linear repair DNA into yeast cells. The protocol is technically facile and requires no special equipment. It can be used in any S. cerevisiae strain, including industrial polyploid isolates. Therefore, this CRISPR-Cas9-based DNA integration protocol is achievable by virtually any yeast genetics and molecular biology laboratory. PMID:27250940

  7. Characterization of Encapsulated Berberine in Yeast Cells of Saccharomyces cerevisiae

    PubMed Central

    Salari, Roshanak; Rajabi, Omid; Khashyarmanesh, Zahra; Fathi Najafi, Mohsen; Fazly Bazzaz, BiBi Sedigheh

    2015-01-01

    Berberine was loaded in yeast cells of Saccharomyces cerevisiaeas a novel pharmaceutical carrier to improve the treatment ofmany diseases. The yeast-encapsulated active materialsshowedhigh stability and bioavailability due to the enhanced solubility and sustained releasing. In this study, different characteristics of prepared berberine loaded yeast cells (loading capacity, release kinetic order, MIC and stability) were evaluatedby different analytical methods (fluorescence spectroscopy, HPLC and SEM).The loading capacity was about 78% ± 0.6%.Berberine release patterns of microcapsules happened in two different stages and followed by zero and first-order kinetic,respectively. About 99% of all active material released during 34 h. MIC was improved by berberine loaded microcapsules in comparison withberberine powder. The microcapsules were completely stable. Berberine loaded Sac. Cerevisiae could be considered as a favorite sustained release drug delivery system. The yeast would be applied as an efficient carrier to improve various properties of different active materials. PMID:26664393

  8. Industrial Systems Biology of Saccharomyces cerevisiae Enables Novel Succinic Acid Cell Factory

    PubMed Central

    Otero, José Manuel; Cimini, Donatella; Patil, Kiran R.; Poulsen, Simon G.; Olsson, Lisbeth; Nielsen, Jens

    2013-01-01

    Saccharomyces cerevisiae is the most well characterized eukaryote, the preferred microbial cell factory for the largest industrial biotechnology product (bioethanol), and a robust commerically compatible scaffold to be exploitted for diverse chemical production. Succinic acid is a highly sought after added-value chemical for which there is no native pre-disposition for production and accmulation in S. cerevisiae. The genome-scale metabolic network reconstruction of S. cerevisiae enabled in silico gene deletion predictions using an evolutionary programming method to couple biomass and succinate production. Glycine and serine, both essential amino acids required for biomass formation, are formed from both glycolytic and TCA cycle intermediates. Succinate formation results from the isocitrate lyase catalyzed conversion of isocitrate, and from the α-keto-glutarate dehydrogenase catalyzed conversion of α-keto-glutarate. Succinate is subsequently depleted by the succinate dehydrogenase complex. The metabolic engineering strategy identified included deletion of the primary succinate consuming reaction, Sdh3p, and interruption of glycolysis derived serine by deletion of 3-phosphoglycerate dehydrogenase, Ser3p/Ser33p. Pursuing these targets, a multi-gene deletion strain was constructed, and directed evolution with selection used to identify a succinate producing mutant. Physiological characterization coupled with integrated data analysis of transcriptome data in the metabolically engineered strain were used to identify 2nd-round metabolic engineering targets. The resulting strain represents a 30-fold improvement in succinate titer, and a 43-fold improvement in succinate yield on biomass, with only a 2.8-fold decrease in the specific growth rate compared to the reference strain. Intuitive genetic targets for either over-expression or interruption of succinate producing or consuming pathways, respectively, do not lead to increased succinate. Rather, we demonstrate how

  9. Architecture and Biosynthesis of the Saccharomyces cerevisiae Cell Wall

    PubMed Central

    Orlean, Peter

    2012-01-01

    The wall gives a Saccharomyces cerevisiae cell its osmotic integrity; defines cell shape during budding growth, mating, sporulation, and pseudohypha formation; and presents adhesive glycoproteins to other yeast cells. The wall consists of β1,3- and β1,6-glucans, a small amount of chitin, and many different proteins that may bear N- and O-linked glycans and a glycolipid anchor. These components become cross-linked in various ways to form higher-order complexes. Wall composition and degree of cross-linking vary during growth and development and change in response to cell wall stress. This article reviews wall biogenesis in vegetative cells, covering the structure of wall components and how they are cross-linked; the biosynthesis of N- and O-linked glycans, glycosylphosphatidylinositol membrane anchors, β1,3- and β1,6-linked glucans, and chitin; the reactions that cross-link wall components; and the possible functions of enzymatic and nonenzymatic cell wall proteins. PMID:23135325

  10. Molecular basis of cell integrity and morphogenesis in Saccharomyces cerevisiae.

    PubMed Central

    Cid, V J; Durán, A; del Rey, F; Snyder, M P; Nombela, C; Sánchez, M

    1995-01-01

    In fungi and many other organisms, a thick outer cell wall is responsible for determining the shape of the cell and for maintaining its integrity. The budding yeast Saccharomyces cerevisiae has been a useful model organism for the study of cell wall synthesis, and over the past few decades, many aspects of the composition, structure, and enzymology of the cell wall have been elucidated. The cell wall of budding yeasts is a complex and dynamic structure; its arrangement alters as the cell grows, and its composition changes in response to different environmental conditions and at different times during the yeast life cycle. In the past few years, we have witnessed a profilic genetic and molecular characterization of some key aspects of cell wall polymer synthesis and hydrolysis in the budding yeast. Furthermore, this organism has been the target of numerous recent studies on the topic of morphogenesis, which have had an enormous impact on our understanding of the intracellular events that participate in directed cell wall synthesis. A number of components that direct polarized secretion, including those involved in assembly and organization of the actin cytoskeleton, secretory pathways, and a series of novel signal transduction systems and regulatory components have been identified. Analysis of these different components has suggested pathways by which polarized secretion is directed and controlled. Our aim is to offer an overall view of the current understanding of cell wall dynamics and of the complex network that controls polarized growth at particular stages of the budding yeast cell cycle and life cycle. PMID:7565410

  11. [Construction of Saccharomyces cerevisiae cell factories for lycopene production].

    PubMed

    Shi, Ming-Yu; Liu Yi; Wang, Dong; Lu, Fu-Ping; Huang, Lu-Qi; Dai, Zhu-Bo; Zhang, Xue-Li

    2014-10-01

    For microbial production of lycopene, the lycopene synthetic genes from Pantoea agglomerans were integrated into Saccharomyces cerevisiae strain BY4742, to obtain strain ZD-L-000 for production of 0.17 mg · L(-1) lycopene. Improving supplies of isoprenoid precursors was then investigated for increasing lycopene production. Four key genes were chosen to be overexpressed, inclu- ding truncated 3-hydroxy-3-methylglutaryl-CoA reductase gene (tHMG1), which is the major rate-limiting enzyme in the mevalonate (MVA) pathway, a mutated global regulatory factor gene (upc2.1), a fusion gene of FPP synthase (ERG20) and endogenous GGPP synthase (BTS1), which is a key enzyme in the diterpenoid synthetic pathway, and GGPP synthase gene (SaGGPS) from Sulfolobus acidocaldarius. Over-expression of upc2.1 could not improve lycopene production, while over-expression of tHMGI , BTS1-ERG20 and SaGGPS genes led to 2-, 16. 9- and20. 5-fold increase of lycopene production, respectively. In addition, three effective genes, tHMG1, BTS1-ERG20 and SaGGPS, were integrated into rDNA sites of ZD-L-000, resulting in strain ZD-L-201 for production of 13.23 mg · L(-1) lycopene, which was 77-fold higher than that of the parent strain. Finally, two-phase extractive fermentation was performed. The titer of lycopene increased 10-fold to 135.21 mg · L(-1). The engineered yeast strains obtained in this work provided the basis for fermentative production of lycopene. PMID:25751950

  12. Carbon nanotube dispersed conductive network for microbial fuel cells

    NASA Astrophysics Data System (ADS)

    Matsumoto, S.; Yamanaka, K.; Ogikubo, H.; Akasaka, H.; Ohtake, N.

    2014-08-01

    Microbial fuel cells (MFCs) are promising devices for capturing biomass energy. Although they have recently attracted considerable attention, their power densities are too low for practical use. Increasing their electrode surface area is a key factor for improving the performance of MFC. Carbon nanotubes (CNTs), which have excellent electrical conductivity and extremely high specific surface area, are promising materials for electrodes. However, CNTs are insoluble in aqueous solution because of their strong intertube van der Waals interactions, which make practical use of CNTs difficult. In this study, we revealed that CNTs have a strong interaction with Saccharomyces cerevisiae cells. CNTs attach to the cells and are dispersed in a mixture of water and S. cerevisiae, forming a three-dimensional CNT conductive network. Compared with a conventional two-dimensional electrode, such as carbon paper, the three-dimensional conductive network has a much larger surface area. By applying this conductive network to MFCs as an anode electrode, power density is increased to 176 μW/cm2, which is approximately 25-fold higher than that in the case without CNTs addition. Maximum current density is also increased to approximately 8-fold higher. These results suggest that three-dimensional CNT conductive network contributes to improve the performance of MFC by increasing surface area.

  13. Kinetics of Saccharomyces cerevisiae elimination from the intestines of human volunteers and effect of this yeast on resistance to microbial colonization in gnotobiotic mice.

    PubMed Central

    Pecquet, S; Guillaumin, D; Tancrede, C; Andremont, A

    1991-01-01

    When healthy volunteers were given a daily dose of 3 x 10(8) life-dehydrated Saccharomyces cerevisiae cells for 5 days, the volunteers excreted 10(5) living yeast cells per g of feces at first, but the yeast cells disappeared within 5 days of the end of treatment. In gnotobiotic mice, S. cerevisiae administered alone colonized the intestinal tract but did not interfere with previous or subsequent colonization by a variety of potentially enteropathogenic microorganisms. When these microorganisms were present, the intestinal counts of S. cerevisiae were greatly reduced. PMID:1746964

  14. Effects of organic and inorganic additives on flotation recovery of washed cells of Saccharomyces cerevisiae resuspended in water.

    PubMed

    DeSousa, Sandro Rogério; Laluce, Cecilia; Jafelicci, Miguel

    2006-03-01

    Separation of microbial cells by flotation recovery is usually carried out in industrial reactors or wastewater treatment systems, which contain a complex mixture of microbial nutrients and excretion products. In the present study, the separation of yeast cells by flotation recovery was carried out using a simple flotation recovery systems containing washed yeast cells resuspended in water in order to elucidate the effects of additives (defined amounts of organic and inorganic acids, ethanol, surfactants and sodium chloride) on the cellular interactions at interfaces (cell/aqueous phase and cell/air bubble). When sodium chloride, organic acids (notably propionic, succinic and acetic acids) and organic surfactants (sodium dodecyl sulphate (SDS), cetyltrimethylammonium bromide (CTAB) and Nonidet P40) were added to the flotation recovery system, significant increases in the cell recovery of yeast hydrophobic cells (Saccharomyces cerevisiae, strain FLT-01) were observed. The association of ethanol to acetic acid solution (a minor by-product of alcoholic fermentation) in the flotation recovery system, containing washed cells of strain FLT-01 resuspended in water, leading to an increased flotation recovery at pH 5.5. Thus, the association among products of the cellular metabolism (e.g., ethanol and acetic acid) can improve yeast cell recovery by flotation recovery. PMID:16500092

  15. Electrophysiology in the eukaryotic model cell Saccharomyces cerevisiae.

    PubMed

    Bertl, A; Bihler, H; Kettner, C; Slayman, C L

    1998-11-01

    Since the mid-1980s, use of the budding yeast, Saccharomyces cerevisiae, for expression of heterologous (foreign) genes and proteins has burgeoned for several major purposes, including facile genetic manipulation, large-scale production of specific proteins, and preliminary functional analysis. Expression of heterologous membrane proteins in yeast has not kept pace with expression of cytoplasmic proteins for two principal reasons: (1) although plant and fungal proteins express and function easily in yeast membranes, animal proteins do not, at least yet; and (2) the yeast plasma membrane is generally regarded as a difficult system to which to apply the standard electrophysiological techniques for detailed functional analysis of membrane proteins. Especially now, since completion of the genome-sequencing project for Saccharomyces, yeast membranes themselves can be seen as an ample source of diverse membrane proteins - including ion channels, pumps, and cotransporters - which lend themselves to electrophysiological analysis, and specifically to patch-clamping. Using some of these native proteins for assay, we report systematic methods to prepare both the yeast plasma membrane and the yeast vacuolar membrane (tonoplast) for patch-clamp experiments. We also describe optimized ambient conditions - such as electrode preparation, buffer solutions, and time regimens - which facilitate efficient patch recording from Saccharomyces membranes. There are two main keys to successful patch-clamping with Saccharomyces. The first is patience; the second is scrupulous cleanliness. Large cells, such as provided by polyploid strains, are also useful in yeast patch recording, especially while the skill required for gigaseal formation is being learned. Cleanliness is aided by (1) osmotic extrusion of protoplasts, after minimal digestion of yeast walls; (2) use of a rather spare suspension of protoplasts in the recording chamber; (3) maintenance of continuous chamber perfusion prior to

  16. Real time, in situ observation of the photocatalytic inactivation of Saccharomyces cerevisiae cells.

    PubMed

    Zhang, Jingtao; Wang, Xiaoxin; Li, Qi; Shang, Jian Ku

    2015-04-01

    An in situ microscopy technique was developed to observe in real time the photocatalytic inactivation process of Saccharomyces cerevisiae (S. cerevisiae) cells by palladium-modified nitrogen-doped titanium oxide (TiON/PdO) under visible light illumination. The technique was based on building a photocatalytic micro-reactor on the sample stage of a fluorescence/phase contrast microscopy capable of simultaneously providing the optical excitation to activate the photocatalyst in the micro-reactor and the illumination to acquire phase contrast images of the cells undergoing the photocatalytic inactivation process. Using TiON/PdO as an example, the technique revealed for the first time the vacuolar activities inside S. cerevisiae cells subjected to a visible light photocatalytic inactivation. The vacuoles responded to the photocatalytic attack by the first expansion of the vacuolar volume and then contraction, before the vacuole disappeared and the cell structure collapsed. Consistent with the aggregate behavior observed from the cell culture experiments, the transition in the vacuolar volume provided clear evidence that photocatalytic disinfection of S. cerevisiae cells started with an initiation period in which cells struggled to offset the photocatalytic damage and moved rapidly after the photocatalytic damage overwhelmed the defense mechanisms of the cells against oxidative attack. PMID:25686929

  17. A novel technique to evaluate interactions between Saccharomyces cerevisiae cell wall and mycotoxins: application to zearalenone.

    PubMed

    Yiannikouris, Alexandros; Poughon, Laurent; Cameleyre, Xavier; Dussap, Claude-Gilles; François, Jean; Bertin, Gérard; Jouany, Jean-Pierre

    2003-05-01

    Three models based on sigmoidal plotting were tested for their ability to describe zearalenone adsorption on Saccharomyces cerevisiae cell walls in vitro. All three models closely fitted the experimental data, but Hill's equation gave the most accurate parameters, and provided information on the physical and chemical mechanisms involved in the adsorption of mycotoxin on yeast cell walls. PMID:12882008

  18. Bioconversion of lactose/whey to fructose diphosphate with recombinant Saccharomyces cerevisiae cells

    SciTech Connect

    Compagno, C.; Tura, A.; Ranzi, B.M.; Martegani, E. )

    1993-07-01

    Genetically engineered Saccharomyces cerevisiae strains that express Escherichia coli [beta]-galactosidase gene are able to bioconvert lactose or whey into fructose-1,6-diphosphate (FDP). High FDP yields from whey were obtained with an appropriate ratio between cell concentration and inorganic phosphate. The biomass of transformed cells can be obtained from different carbon sources, according to the expression vector bearing the lacZ gene. The authors showed that whey can be used as the carbon source for S. cerevisiae growth and as the substrate for bioconversion to fructose diphosphate.

  19. Microbial Cell Factories for Diol Production.

    PubMed

    Sabra, W; Groeger, C; Zeng, An-Ping

    2016-01-01

    Diols are compounds with two hydroxyl groups and have a wide range of appealing applications as chemicals and fuels. In particular, five low molecular diol compounds, namely 1,3-propanediol (1,3-PDO), 1,2-propanediol (1,2-PDO), 2,3-butanediol (2,3-BDO), 1,3-butanediol (1,3-BDO), and 1,4-butanediol (1,4-BDO), can be biotechnologically produced by direct microbial bioconversion of renewable materials. In this review, we summarize recent developments in the microbial production of diols, especially regarding the engineering of typical microbial strains as cell factory and the development of corresponding bioconversion processes. PMID:26475465

  20. Lead sulfide nanoparticles increase cell wall chitin content and induce apoptosis in Saccharomyces cerevisiae.

    PubMed

    Sun, Meiqing; Yu, Qilin; Hu, Mengyuan; Hao, Zhenwei; Zhang, Chengdong; Li, Mingchun

    2014-05-30

    Although there have been numerous studies on bacterial toxicity, the cytotoxicity of nanoparticles toward fungi remains poorly understood. We investigated the toxicity of various sizes of lead sulfide particles against the important model fungus, Saccharomyces cerevisiae. The smallest particle exerted the highest toxicity, inhibiting cell growth and decreasing cell viability, likely reflecting reduced sedimentation and persistent cell wall attack. In response to cell wall stress, S. cerevisiae showed an increase in the cell wall chitin content and the overexpression of FKS2 and PRM5, two genes of the cell wall integrity signaling pathway. Cell wall stress increased the concentration of intracellular reactive oxygen species, leading to mitochondrial dysfunction and cell apoptosis. The contribution of dissolved lead ions to the overall toxicity was negligible. These findings provide the first demonstration of the physiological protective response of a fungus toward nanoparticles, thereby contributing useful information to the assessment of the environmental impact of metal nanoparticles. PMID:24704549

  1. Why should cell biologists study microbial pathogens?

    PubMed Central

    Welch, Matthew D.

    2015-01-01

    One quarter of all deaths worldwide each year result from infectious diseases caused by microbial pathogens. Pathogens infect and cause disease by producing virulence factors that target host cell molecules. Studying how virulence factors target host cells has revealed fundamental principles of cell biology. These include important advances in our understanding of the cytoskeleton, organelles and membrane-trafficking intermediates, signal transduction pathways, cell cycle regulators, the organelle/protein recycling machinery, and cell-death pathways. Such studies have also revealed cellular pathways crucial for the immune response. Discoveries from basic research on the cell biology of pathogenesis are actively being translated into the development of host-targeted therapies to treat infectious diseases. Thus there are many reasons for cell biologists to incorporate the study of microbial pathogens into their research programs. PMID:26628749

  2. Biosorption of water-soluble dyes on magnetically modified Saccharomyces cerevisiae subsp. uvarum cells.

    PubMed

    Safaríková, M; Ptácková, L; Kibriková, I; Safarík, I

    2005-05-01

    Brewer's yeast (bottom yeast, Saccharomyces cerevisiae subsp. uvarum) cells were magnetically modified using water based magnetic fluid stabilized with perchloric acid. Magnetically modified yeast cells efficiently adsorbed various water soluble dyes. The dyes adsorption can be described by the Langmuir adsorption model. The maximum adsorption capacity of the magnetic cells differed substantially for individual dyes; the highest value was found for aniline blue (approx. 220 mg per g of dried magnetic adsorbent). PMID:15811411

  3. Molecular mechanisms of Saccharomyces cerevisiae stress adaptation and programmed cell death in response to acetic acid

    PubMed Central

    Giannattasio, Sergio; Guaragnella, Nicoletta; Ždralević, Maša; Marra, Ersilia

    2013-01-01

    Beyond its classical biotechnological applications such as food and beverage production or as a cell factory, the yeast Saccharomyces cerevisiae is a valuable model organism to study fundamental mechanisms of cell response to stressful environmental changes. Acetic acid is a physiological product of yeast fermentation and it is a well-known food preservative due to its antimicrobial action. Acetic acid has recently been shown to cause yeast cell death and aging. Here we shall focus on the molecular mechanisms of S. cerevisiae stress adaptation and programmed cell death in response to acetic acid. We shall elaborate on the intracellular signaling pathways involved in the cross-talk of pro-survival and pro-death pathways underlying the importance of understanding fundamental aspects of yeast cell homeostasis to improve the performance of a given yeast strain in biotechnological applications. PMID:23430312

  4. Molecular mechanisms of Saccharomyces cerevisiae stress adaptation and programmed cell death in response to acetic acid.

    PubMed

    Giannattasio, Sergio; Guaragnella, Nicoletta; Zdralević, Maša; Marra, Ersilia

    2013-01-01

    Beyond its classical biotechnological applications such as food and beverage production or as a cell factory, the yeast Saccharomyces cerevisiae is a valuable model organism to study fundamental mechanisms of cell response to stressful environmental changes. Acetic acid is a physiological product of yeast fermentation and it is a well-known food preservative due to its antimicrobial action. Acetic acid has recently been shown to cause yeast cell death and aging. Here we shall focus on the molecular mechanisms of S. cerevisiae stress adaptation and programmed cell death in response to acetic acid. We shall elaborate on the intracellular signaling pathways involved in the cross-talk of pro-survival and pro-death pathways underlying the importance of understanding fundamental aspects of yeast cell homeostasis to improve the performance of a given yeast strain in biotechnological applications. PMID:23430312

  5. Ethanol production from nonsterilized carob pod extract by free and immobilized Saccharomyces cerevisiae cells using fed-batch culture

    SciTech Connect

    Roukas, T. . Dept. of Food Science and Technology)

    1994-02-05

    The production of ethanol from carob pod extract by free and immobilized Saccharomyces cerevisiae cells in batch and fed-batch culture was investigated. Fed-batch culture proved to be a better fermentation system for the production of ethanol than batch culture. In fed-batch culture, both free and immobilized S. cerevisiae cells gave the same maximum concentration of final ethanol at an initial sugar concentration of 300 g/L and F = 167 mL/h. The maximum ethanol productivity was obtained with both free and immobilized cells at a substrate concentration of 300 g/L and F = 334 mL/h. In repeated fed-batch culture, immobilized S. cerevisiae cells gave a higher overall ethanol concentration compared with the free cells. The immobilized S. cerevisiae cells in Ca-alginate beads retained their ability to produce ethanol for 10 days.

  6. Regulation of Cell Wall Biogenesis in Saccharomyces cerevisiae: The Cell Wall Integrity Signaling Pathway

    PubMed Central

    Levin, David E.

    2011-01-01

    The yeast cell wall is a strong, but elastic, structure that is essential not only for the maintenance of cell shape and integrity, but also for progression through the cell cycle. During growth and morphogenesis, and in response to environmental challenges, the cell wall is remodeled in a highly regulated and polarized manner, a process that is principally under the control of the cell wall integrity (CWI) signaling pathway. This pathway transmits wall stress signals from the cell surface to the Rho1 GTPase, which mobilizes a physiologic response through a variety of effectors. Activation of CWI signaling regulates the production of various carbohydrate polymers of the cell wall, as well as their polarized delivery to the site of cell wall remodeling. This review article centers on CWI signaling in Saccharomyces cerevisiae through the cell cycle and in response to cell wall stress. The interface of this signaling pathway with other pathways that contribute to the maintenance of cell wall integrity is also discussed. PMID:22174182

  7. Autonomous, Retrievable, Deep Sea Microbial Fuel Cell

    NASA Astrophysics Data System (ADS)

    Richter, K.

    2014-12-01

    Microbial fuel cells (MFCs) work by providing bacteria in anaerobic sediments with an electron acceptor (anode) that stimulates metabolism of organic matter. The buried anode is connected via control circuitry to a cathode exposed to oxygen in the overlying water. During metabolism, bacteria release hydrogen ions into the sediment and transfer electrons extra-cellularly to the anode, which eventually reduce dissolved oxygen at the cathode, forming water. The open circuit voltage is approximately 0.8 v. The voltage between electrodes is operationally kept at 0.4 v with a potentiastat. The current is chiefly limited by the rate of microbial metabolism at the anode. The Office of Naval Research has encouraged development of microbial fuel cells in the marine environment at a number of academic and naval institutions. Earlier work in shallow sediments of San Diego Bay showed that the most important environmental parameters that control fuel cell power output in San Diego Bay were total organic carbon in the sediment and seasonal water temperature. Current MFC work at SPAWAR includes extension of microbial fuel cell tests to the deep sea environment (>1000 m) and, in parallel, testing microbial fuel cells in the laboratory under deep sea conditions. One question we are asking is whether MFC power output from deep water sediments repressurized and chilled in the laboratory comparable to those measured in situ. If yes, mapping the power potential of deep sea sediments may be made much easier, requiring sediment grabs and lab tests rather than deployment and retrieval of fuel cells. Another question we are asking is whether in situ temperature and total organic carbon in the deep sea sediment can predict MFC power. If yes, then we can make use of the large collection of publicly available, deep sea oceanographic measurements to make these predictions, foregoing expensive work at sea. These regressions will be compared to those derived from shallow water measurements.

  8. Interval scanning photomicrography of microbial cell populations.

    NASA Technical Reports Server (NTRS)

    Casida, L. E., Jr.

    1972-01-01

    A single reproducible area of the preparation in a fixed focal plane is photographically scanned at intervals during incubation. The procedure can be used for evaluating the aerobic or anaerobic growth of many microbial cells simultaneously within a population. In addition, the microscope is not restricted to the viewing of any one microculture preparation, since the slide cultures are incubated separately from the microscope.

  9. Single-cell transcriptomics for microbial eukaryotes.

    PubMed

    Kolisko, Martin; Boscaro, Vittorio; Burki, Fabien; Lynn, Denis H; Keeling, Patrick J

    2014-11-17

    One of the greatest hindrances to a comprehensive understanding of microbial genomics, cell biology, ecology, and evolution is that most microbial life is not in culture. Solutions to this problem have mainly focused on whole-community surveys like metagenomics, but these analyses inevitably loose information and present particular challenges for eukaryotes, which are relatively rare and possess large, gene-sparse genomes. Single-cell analyses present an alternative solution that allows for specific species to be targeted, while retaining information on cellular identity, morphology, and partitioning of activities within microbial communities. Single-cell transcriptomics, pioneered in medical research, offers particular potential advantages for uncultivated eukaryotes, but the efficiency and biases have not been tested. Here we describe a simple and reproducible method for single-cell transcriptomics using manually isolated cells from five model ciliate species; we examine impacts of amplification bias and contamination, and compare the efficacy of gene discovery to traditional culture-based transcriptomics. Gene discovery using single-cell transcriptomes was found to be comparable to mass-culture methods, suggesting single-cell transcriptomics is an efficient entry point into genomic data from the vast majority of eukaryotic biodiversity. PMID:25458215

  10. Microbial fuel cells for biosensor applications.

    PubMed

    Yang, Huijia; Zhou, Minghua; Liu, Mengmeng; Yang, Weilu; Gu, Tingyue

    2015-12-01

    Microbial fuel cells (MFCs) face major hurdles for real-world applications as power generators with the exception of powering small sensor devices. Despite tremendous improvements made in the last two decades, MFCs are still too expensive to build and operate and their power output is still too small. In view of this, in recently years, intensive researches have been carried out to expand the applications into other areas such as acid and alkali production, bioremediation of aquatic sediments, desalination and biosensors. Unlike power applications, MFC sensors have the immediate prospect to be practical. This review covers the latest developments in various proposed biosensor applications using MFCs including monitoring microbial activity, testing biochemical oxygen demand, detection of toxicants and detection of microbial biofilms that cause biocorrosion. PMID:26272393

  11. Endomitotic effect of a cell cycle mutation of Saccharomyces cerevisiae

    SciTech Connect

    Schild, D.; Ananthaswamy, H.N.; Mortimer, R.K.

    1981-03-01

    A recessive temperature-sensitive mutation of Saccharomyces cerevisiae has been isolated and shown to cause an increase in ploidy in both haploids and diploids. Genetic analysis revealed that the strain carrying the mutation was an aa diploid, although MNNG mutagenesis had been done on an a haploid strain. When the mutant strain was crossed with an ..cap alpha cap alpha.. diploid and the resultant tetraploid sporulated, some of the meiotic progeny of this tetraploid were themselves tetraploid, as shown by both genetic analysis and DNA measurements, instead of diploid as expected of tetraploid meiosis. The ability of these tetraploids to continue to produce tetraploid meiotic progeny was followed for four generations. It was found that tetraploidization was independent of sporulation temperature, but was dependent on the temperature of germination and the growth of the spores. Increase in ploidy occurred when the spores were germinated and grown at 30/sup 0/, but did not occur at 23/sup 0/. Two cycles of sporulation and growth at 23/sup 0/ resulted in haploids, which were shown to diploidize within 24 hr when grown at 30/sup 0/.

  12. Brazilian propolis protects Saccharomyces cerevisiae cells against oxidative stress

    PubMed Central

    de Sá, Rafael A.; de Castro, Frederico A.V.; Eleutherio, Elis C.A.; de Souza, Raquel M.; da Silva, Joaquim F.M.; Pereira, Marcos D.

    2013-01-01

    Propolis is a natural product widely used for humans. Due to its complex composition, a number of applications (antimicrobial, antiinflammatory, anesthetic, cytostatic and antioxidant) have been attributed to this substance. Using Saccharomyces cerevisiae as a eukaryotic model we investigated the mechanisms underlying the antioxidant effect of propolis from Guarapari against oxidative stress. Submitting a wild type (BY4741) and antioxidant deficient strains (ctt1Δ, sod1Δ, gsh1Δ, gtt1Δ and gtt2Δ) either to 15 mM menadione or to 2 mM hydrogen peroxide during 60 min, we observed that all strains, except the mutant sod1Δ, acquired tolerance when previously treated with 25 μg/mL of alcoholic propolis extract. Such a treatment reduced the levels of ROS generation and of lipid peroxidation, after oxidative stress. The increase in Cu/Zn-Sod activity by propolis suggests that the protection might be acting synergistically with Cu/Zn-Sod. PMID:24516431

  13. From microbial communities to cells

    NASA Technical Reports Server (NTRS)

    Margulis, L.

    1985-01-01

    The eukraotic cell, the unit of structure of protoctists, plants, fungi, and animals, is not at all homologous to prokaryotic cells. Instead the eukaryotic cell is homologous to communities of microorganisms such as those of the sulfuretum. This research is based on the hypothesis that at least four different interacting community members entered the original associations that, when stabilized, led to the emergence of eukaryotic cells. These are: (1) host nucleocytoplasm (thermoplasma like archaebacteria); (2) mitochrondria (paracoccus or bdellovibryo like respiring bacteria; and (3) plastids (cyanobacteria) and undulipodia. Tubulin like protein was found in the free living spirochete Spirochaeta bajacaliforniensis and in several other spirochetes. The amino acid sequence was to see if the spirochete protein is homologous to the tubulin of undulipodial and mitotic spindle microtubules.

  14. Effects of dietary changes and yeast culture (Saccharomyces cerevisiae) on rumen microbial fermentation of Holstein heifers.

    PubMed

    Moya, D; Calsamiglia, S; Ferret, A; Blanch, M; Fandiño, J I; Castillejos, L; Yoon, I

    2009-09-01

    The effects of a dietary challenge to induce digestive upsets and supplementation with yeast culture on rumen microbial fermentation were studied using 12 Holstein heifers (277 +/- 28 kg of BW) fitted with a ruminal cannula, in a crossover design with 2 periods of 5 wk. In each period, after 3 wk of adaptation to a 100% forage diet, the dietary challenge consisted of increasing the amount of grain at a rate of 2.5 kg/d (as-fed basis) over a period of 4 d, until a 10:90 forage:concentrate diet was reached, and then it was maintained for 10 d. Between periods, animals were fed again the 100% forage diet without any treatment for 1 wk as a wash-out period. Treatments started the first day of each period, and they were a control diet (CL) or the same diet with addition of yeast culture (YC, Diamond V XPCLS). Digestive upsets were determined by visual observation of bloat or by a reduction in feed intake (as-fed basis) of 50% or more compared with intake on the previous day. Feed intake was determined daily at 24-h intervals during the adaptation period and daily at 2, 6, and 12 h postfeeding during the dietary challenge. Ruminal liquid samples were collected daily during the dietary challenge to determine ruminal pH at 0, 3, 6, and 12 h postfeeding, and total and individual VFA, lactic acid, ammonia-N, and rumen fluid viscosity at 0 and 6 h postfeeding. The 16s rRNA gene copies of Streptococcus bovis and Megasphaera elsdenii were determined by quantitative PCR. Foam height and strength of the rumen fluid were also determined the day after the digestive upset to evaluate potential foam production. A total of 20 cases (83.3%) of digestive upsets were recorded in both periods during the dietary challenge, all diagnosed due to a reduction in feed intake. Rumen fermentation profile at 0 h on the digestive upset day was characterized by low ruminal pH, which remained under 6.0 for 18 h, accompanied by elevated total VFA concentration and, in some cases, by elevated lactate

  15. Life cycle assessment of high-rate anaerobic treatment, microbial fuel cells, and microbial electrolysis cells.

    PubMed

    Foley, Jeffrey M; Rozendal, René A; Hertle, Christopher K; Lant, Paul A; Rabaey, Korneel

    2010-05-01

    Existing wastewater treatment options are generally perceived as energy intensive and environmentally unfriendly. Much attention has been focused on two new approaches in the past years, (i) microbial fuel cells and (ii) microbial electrolysis cells, which directly generate electrical current or chemical products, respectively, during wastewater treatment. These systems are commonly denominated as bioelectrochemical systems, and a multitude of claims have been made in the past regarding the environmental impact of these treatment options. However, an in-depth study backing these claims has not been performed. Here, we have conducted a life cycle assessment (LCA) to compare the environmental impact of three industrial wastewater treatment options, (i) anaerobic treatment with biogas generation, (ii) a microbial fuel cell treatment, with direct electricity generation, and (iii) a microbial electrolysis cell, with hydrogen peroxide production. Our analysis showed that a microbial fuel cell does not provide a significant environmental benefit relative to the "conventional" anaerobic treatment option. However, a microbial electrolysis cell provides significant environmental benefits through the displacement of chemical production by conventional means. Provided that the target conversion level of 1000 A.m(-3) can be met, the decrease in greenhouse gas emissions and other environmentally harmful emissions (e.g., aromatic hydrocarbons) of the microbial electrolysis cell will be a key driver for the development of an industrial standard for this technology. Evidently, this assessment is highly dependent on the underlying assumptions, such as the used reactor materials and target performance. This provides a challenge and an opportunity for researchers in the field to select and develop appropriate and environmentally benign materials of construction, as well as demonstrate the required 1000 A.m(-3) performance at pilot and full scale. PMID:20356090

  16. Microbial fuel cell with improved anode

    DOEpatents

    Borole, Abhijeet P.

    2010-04-13

    The present invention relates to a method for preparing a microbial fuel cell, wherein the method includes: (i) inoculating an anodic liquid medium in contact with an anode of the microbial fuel cell with one or more types of microorganisms capable of functioning by an exoelectrogenic mechanism; (ii) establishing a biofilm of the microorganisms on and/or within the anode along with a substantial absence of planktonic forms of the microorganisms by substantial removal of the planktonic microorganisms during forced flow and recirculation conditions of the anodic liquid medium; and (iii) subjecting the microorganisms of the biofilm to a growth stage by incorporating one or more carbon-containing nutritive compounds in the anodic liquid medium during biofilm formation or after biofilm formation on the anode has been established.

  17. Effect of Saccharomyces cerevisiae on alfalfa nutrient degradation characteristics and rumen microbial populations of steers fed diets with different concentrate-to-forage ratios

    PubMed Central

    2014-01-01

    Live yeast (Saccharomyces cerevisiae) constitutes an effective additive for animal production; its probiotic effect may be related to the concentrate-to-forage ratio (CTFR). The objective of this study was to assess the effects of S. cerevisiae (SC) on fiber degradation and rumen microbial populations in steers fed diets with different levels of dietary concentrate. Ten Simmental × Local crossbred steers (450 ± 50 kg BW) were assigned to a control group or an SC group. Both groups were fed the same basal diet but the SC group received SC supplementation (8 × 109 cfu/h/d through the ruminal fistula) following a two-period crossover design. Each period consisted of four phases, each of which lasted 17 d: 10 d for dietary adaptation, 6 d for degradation study, and 1 d for rumen sample collection. From the 1st to the 4th phase, steers were fed in a stepwise fashion with increasing CTFRs, i.e., 30:70, 50:50, 70:30, and 90:10. The kinetics of dry matter and fiber degradation of alfalfa pellets were evaluated; the rumen microbial populations were detected using real-time PCR. The results revealed no significant (P > 0.05) interactions between dietary CTFR and SC for most parameters. Dietary CTFR had a significant effect (P < 0.01) on degradation characteristics of alfalfa pellets and the copies of rumen microorganism; the increasing concentrate level resulted in linear, quadratic or cubic variation trend for these parameters. SC supplementation significantly (P < 0.05) affected dry matter (DM) and neutral detergent fiber (NDF) degradation rates (cDM, cNDF) and NDF effective degradability (EDNDF). Compared with the control group, there was an increasing trend of rumen fungi and protozoa in SC group (P < 0.1); copies of total bacteria in SC group were significantly higher (P < 0.05). Additionally, percentage of Ruminobacter amylophilus was significantly lower (P < 0.05) but percentage of Selenomonas ruminantium was significantly

  18. Effect of Saccharomyces cerevisiae on alfalfa nutrient degradation characteristics and rumen microbial populations of steers fed diets with different concentrate-to-forage ratios.

    PubMed

    Ding, Gengzhi; Chang, Ying; Zhao, Liping; Zhou, Zhenming; Ren, Liping; Meng, Qingxiang

    2014-01-01

    Live yeast (Saccharomyces cerevisiae) constitutes an effective additive for animal production; its probiotic effect may be related to the concentrate-to-forage ratio (CTFR). The objective of this study was to assess the effects of S. cerevisiae (SC) on fiber degradation and rumen microbial populations in steers fed diets with different levels of dietary concentrate. Ten Simmental × Local crossbred steers (450 ± 50 kg BW) were assigned to a control group or an SC group. Both groups were fed the same basal diet but the SC group received SC supplementation (8 × 10(9) cfu/h/d through the ruminal fistula) following a two-period crossover design. Each period consisted of four phases, each of which lasted 17 d: 10 d for dietary adaptation, 6 d for degradation study, and 1 d for rumen sample collection. From the 1(st) to the 4(th) phase, steers were fed in a stepwise fashion with increasing CTFRs, i.e., 30:70, 50:50, 70:30, and 90:10. The kinetics of dry matter and fiber degradation of alfalfa pellets were evaluated; the rumen microbial populations were detected using real-time PCR. The results revealed no significant (P > 0.05) interactions between dietary CTFR and SC for most parameters. Dietary CTFR had a significant effect (P < 0.01) on degradation characteristics of alfalfa pellets and the copies of rumen microorganism; the increasing concentrate level resulted in linear, quadratic or cubic variation trend for these parameters. SC supplementation significantly (P < 0.05) affected dry matter (DM) and neutral detergent fiber (NDF) degradation rates (c DM, c NDF) and NDF effective degradability (EDNDF). Compared with the control group, there was an increasing trend of rumen fungi and protozoa in SC group (P < 0.1); copies of total bacteria in SC group were significantly higher (P < 0.05). Additionally, percentage of Ruminobacter amylophilus was significantly lower (P < 0.05) but percentage of Selenomonas ruminantium was

  19. Impact of Photocatalysis on Fungal Cells: Depiction of Cellular and Molecular Effects on Saccharomyces cerevisiae

    PubMed Central

    Thabet, Sana; Simonet, France; Lemaire, Marc; Guillard, Chantal

    2014-01-01

    We have investigated the antimicrobial effects of photocatalysis on the yeast model Saccharomyces cerevisiae. To accurately study the antimicrobial mechanisms of the photocatalytic process, we focused our investigations on two questions: the entry of the nanoparticles in treated cells and the fate of the intracellular environment. Transmission electronic microscopy did not reveal any entry of nanoparticles within the cells, even for long exposure times, despite degradation of the cell wall space and deconstruction of cellular compartments. In contrast to proteins located at the periphery of the cells, intracellular proteins did not disappear uniformly. Disappearance or persistence of proteins from the pool of oxidized intracellular isoforms was not correlated to their functions. Altogether, our data suggested that photocatalysis induces the establishment of an intracellular oxidative environment. This hypothesis was sustained by the detection of an increased level of superoxide ions (O2°−) in treated cells and by greater cell cultivability for cells expressing oxidant stress response genes during photocatalytic exposure. The increase in intracellular ROS, which was not connected to the entry of nanoparticles within the cells or to a direct contact with the plasma membrane, could be the result of an imbalance in redox status amplified by chain reactions. Moreover, we expanded our study to other yeast and filamentous fungi and pointed out that, in contrast to the laboratory model S. cerevisiae, some environmental strains are very resistant to photocatalysis. This could be related to the cell wall composition and structure. PMID:25261515

  20. Microbial fuel cells: Their application and microbiology

    NASA Astrophysics Data System (ADS)

    He, Zhen

    The energy crisis is an urgent global issue due to the increased consumption of the finite amount of fossil fuel. As a result, looking for alternative energy sources is of critical importance. Microbial fuel cell (MFC) technology can extract electric energy from wastewater, and thus is a sustainable approach to supply energy to our electricity-based society. My research focuses on the development of a suitable MFC reactor for wastewater treatment and the understanding of the microbial function in the MFC process. First, together with colleagues, I have developed a novel MFC reactor, named upflow microbial fuel cell (UMFC), by combining upflow and MFC technologies. The power output from the UMFC was improved by 10-fold after it was modified with a U-shape cathode. The UMFC appears to be a practical reactor for continuous operation, though the output of electric power requires further improvement. In addition, a sediment MFC with a rotating cathode was also developed and its performance was examined. Second, I have adopted a human distal gut anaerobe, Bacteroides thetaiotaomicron, as the model organism to study the role of fermentative bacterium in electricity generation. When B. thetaiotaomicron grew under an applied electric potential, an electric current was generated. GeneChip data indicated that this bacterium did not alter its metabolism during this process. Although B. thetaiotaomicron may not be capable of respiration with an electrode as the electron acceptor, the experiment has demonstrated that fermentative bacteria may play an important role in electricity generation.

  1. Decreased fluidity of cell membranes causes a metal ion deficiency in recombinant Saccharomyces cerevisiae producing carotenoids.

    PubMed

    Liu, Peitong; Sun, Liang; Sun, Yuxia; Shang, Fei; Yan, Guoliang

    2016-04-01

    The genome-wide transcriptional responses of S. cerevisiae to heterologous carotenoid biosynthesis were investigated using DNA microarray analysis. The results show that the genes involved in metal ion transport were specifically up-regulated in the recombinant strain, and metal ions, including Cu(2+), Fe(2+), Mn(2+), and Mg(2+), were deficient in the recombinant strain compared to the ion content of the parent strain. The decrease in metal ions was ascribed to a decrease in cell membrane (CM) fluidity caused by lower levels of unsaturated fatty acids and ergosterol. This was confirmed by the observation that metal ion levels were restored when CM fluidity was increased by supplying linoleic acid. In addition, a 24.3 % increase in the β-carotene concentration was observed. Collectively, our results suggest that heterologous production of carotenoids in S. cerevisiae can induce cellular stress by rigidifying the CM, which can lead to a deficiency in metal ions. Due to the importance of CM fluidity in cellular physiology, maintaining normal CM fluidity might be a potential approach to improving carotenoid production in genetically engineered S. cerevisiae. PMID:26749524

  2. Cell death caused by excision of centromeric DNA from a chromosome in Saccharomyces cerevisiae.

    PubMed

    Miyamoto, Akihiro; Yanamoto, Toshiaki; Matsumoto, Takehiro; Hatano, Takushi; Matsuzaki, Hiroaki

    2013-01-01

    If genetically modified organisms (GMOs) are spread through the natural environment, it might affect the natural environment. To help prevent the spread of GMOs, we examined whether it is possible to introduce conditional lethality by excising centromeric DNA from a chromosome by site-specific recombination in Saccharomyces cerevisiae as model organism. First, we constructed haploid cells in which excision of the centromeric DNA from chromosome IV can occur due to recombinase induced by galactose. By this excision, cell death can occur. In diploid cells, cell death can also occur by excision from both homologous chromosomes IV. Furthermore, cell death can occur in the case of chromosome V. A small number of surviving cells appeared with excision of centromeric DNA, and the diploid showed greater viability than the haploid in both chromosomes IV and V. The surviving cells appeared mainly due to deletion of a recombination target site (RS) from the chromosome. PMID:24018677

  3. Members of the Hsp70 family of proteins in the cell wall of Saccharomyces cerevisiae.

    PubMed Central

    López-Ribot, J L; Chaffin, W L

    1996-01-01

    Western blot (immunoblot) analysis of cell wall and cytosolic extracts obtained from parental and ssa1 and ssa2 single- and double-mutant strains of Saccharomyces cerevisiae showed that the heat shock protein 70 (Hsp70) products of these genes, previously thought to be restricted to the cell interior, are also present in the cell wall. A cell wall location was further confirmed by indirect immunofluorescence with intact cells and biotinylation of extracellular Hsp70. Hsp70s have been implicated in translocation across the membrane and as molecular chaperones, and changes in the profile of cell wall proteins suggested that these proteins may have a similar role in the cell wall. PMID:8755907

  4. Electrochemical detection of intracellular and cell membrane redox systems in Saccharomyces cerevisiae

    NASA Astrophysics Data System (ADS)

    Rawson, Frankie J.; Downard, Alison J.; Baronian, Keith H.

    2014-06-01

    Redox mediators can interact with eukaryote cells at a number of different cell locations. While cell membrane redox centres are easily accessible, the redox centres of catabolism are situated within the cytoplasm and mitochondria and can be difficult to access. We have systematically investigated the interaction of thirteen commonly used lipophilic and hydrophilic mediators with the yeast Saccharomyces cerevisiae. A double mediator system is used in which ferricyanide is the final electron acceptor (the reporter mediator). After incubation of cells with mediators, steady state voltammetry of the ferri/ferrocyanide redox couple allows quantitation of the amount of mediator reduced by the cells. The plateau current at 425 mV vs Ag/AgCl gives the analytical signal. The results show that five of the mediators interact with at least three different trans Plasma Membrane Electron Transport systems (tPMETs), and that four mediators cross the plasma membrane to interact with cytoplasmic and mitochondrial redox molecules. Four of the mediators inhibit electron transfer from S. cerevisiae. Catabolic inhibitors were used to locate the cellular source of electrons for three of the mediators.

  5. Electrochemical detection of intracellular and cell membrane redox systems in Saccharomyces cerevisiae

    PubMed Central

    Rawson, Frankie J.; Downard, Alison J.; Baronian, Keith H.

    2014-01-01

    Redox mediators can interact with eukaryote cells at a number of different cell locations. While cell membrane redox centres are easily accessible, the redox centres of catabolism are situated within the cytoplasm and mitochondria and can be difficult to access. We have systematically investigated the interaction of thirteen commonly used lipophilic and hydrophilic mediators with the yeast Saccharomyces cerevisiae. A double mediator system is used in which ferricyanide is the final electron acceptor (the reporter mediator). After incubation of cells with mediators, steady state voltammetry of the ferri/ferrocyanide redox couple allows quantitation of the amount of mediator reduced by the cells. The plateau current at 425 mV vs Ag/AgCl gives the analytical signal. The results show that five of the mediators interact with at least three different trans Plasma Membrane Electron Transport systems (tPMETs), and that four mediators cross the plasma membrane to interact with cytoplasmic and mitochondrial redox molecules. Four of the mediators inhibit electron transfer from S. cerevisiae. Catabolic inhibitors were used to locate the cellular source of electrons for three of the mediators. PMID:24910017

  6. Shape recognition of microbial cells by colloidal cell imprints

    NASA Astrophysics Data System (ADS)

    Borovička, Josef; Stoyanov, Simeon D.; Paunov, Vesselin N.

    2013-08-01

    We have engineered a class of colloids which can recognize the shape and size of targeted microbial cells and selectively bind to their surfaces. These imprinted colloid particles, which we called ``colloid antibodies'', were fabricated by partial fragmentation of silica shells obtained by templating the targeted microbial cells. We successfully demonstrated the shape and size recognition between such colloidal imprints and matching microbial cells. High percentage of binding events of colloidal imprints with the size matching target particles was achieved. We demonstrated selective binding of colloidal imprints to target microbial cells in a binary mixture of cells of different shapes and sizes, which also resulted in high binding selectivity. We explored the role of the electrostatic interactions between the target cells and their colloid imprints by pre-coating both of them with polyelectrolytes. Selective binding occurred predominantly in the case of opposite surface charges of the colloid cell imprint and the targeted cells. The mechanism of the recognition is based on the amplification of the surface adhesion in the case of shape and size match due to the increased contact area between the target cell and the colloidal imprint. We also tested the selective binding for colloid imprints of particles of fixed shape and varying sizes. The concept of cell recognition by colloid imprints could be used for development of colloid antibodies for shape-selective binding of microbes. Such colloid antibodies could be additionally functionalized with surface groups to enhance their binding efficiency to cells of specific shape and deliver a drug payload directly to their surface or allow them to be manipulated using external fields. They could benefit the pharmaceutical industry in developing selective antimicrobial therapies and formulations.

  7. [Advance in producing higher alcohols by microbial cell factories].

    PubMed

    Liu, Zengran; Zhang, Guangyi

    2013-10-01

    Higher alcohols have a high energy density, low hygroscopicity and can be mixed with gasoline at any ratio. It is the trend to replace fossil fuels with biofuels produced via microbial fermentation of renewable resources. We reviewed the progress in the development of engineered Saccharomyces cerevisiae and Escherichia coli that can produce higher alcohols, as well as the related technology platforms. We mainly focused on the construction of CoA-dependent pathways and alpha-keto acid mediated non-fermentative pathways, analyzed their respective characteristics, and summarized the construction strategies. The problems to be solved and future research direction were also discussed. PMID:24432657

  8. Amyloid-like properties of Saccharomyces cerevisiae cell wall glucantransferase Bgl2p

    PubMed Central

    Plotnikova, Tatyana A; Gorkovskii, Anton A; Selyakh, Irina O; Galzitskaya, Oxana V; Bezsonov, Evgeniy E; Gellissen, Gerd; Kulaev, Igor S

    2008-01-01

    Glucantransferase Bgl2p is a major conserved cell wall constituent described for a wide range of yeast species. In the baker's yeast Saccharomyces cerevisiae it is the only non-covalently bound cell wall protein that cannot be released from cell walls by sequential SDS and trypsin treatment. It contains seven amyloidogenic determinants. Circular dichroism analysis and fluorescence spectroscopy with thioflavin T indicate the presence of β-sheet structures in Bgl2p isolates. Bgl2p forms fibrils, a process that is enforced in the presence of other cell wall components. Thus the data obtained is the first evidence for amyloid-like properties of yeast cell wall protein—glucantransferase Bgl2p. PMID:19098439

  9. Bimolecular fluorescence complementation (BiFC) technique in yeast Saccharomyces cerevisiae and mammalian cells.

    PubMed

    Weber-Boyvat, Marion; Li, Shiqian; Skarp, Kari-Pekka; Olkkonen, Vesa M; Yan, Daoguang; Jäntti, Jussi

    2015-01-01

    Visualization of protein-protein interactions in vivo offers a powerful tool to resolve spatial and temporal aspects of cellular functions. The bimolecular fluorescence complementation (BiFC) makes use of nonfluorescent fragments of green fluorescent protein or its variants that are added as "tags" to target proteins under study. Only upon target protein interaction is a fluorescent protein complex assembled, and the site of interaction can be monitored by microscopy. In this chapter, we describe the method and tools for the use of BiFC in the yeast Saccharomyces cerevisiae and in mammalian cells. PMID:25702124

  10. The final cut: cell polarity meets cytokinesis at the bud neck in S. cerevisiae.

    PubMed

    Juanes, Maria Angeles; Piatti, Simonetta

    2016-08-01

    Cell division is a fundamental but complex process that gives rise to two daughter cells. It includes an ordered set of events, altogether called "the cell cycle", that culminate with cytokinesis, the final stage of mitosis leading to the physical separation of the two daughter cells. Symmetric cell division equally partitions cellular components between the two daughter cells, which are therefore identical to one another and often share the same fate. In many cases, however, cell division is asymmetrical and generates two daughter cells that differ in specific protein inheritance, cell size, or developmental potential. The budding yeast Saccharomyces cerevisiae has proven to be an excellent system to investigate the molecular mechanisms governing asymmetric cell division and cytokinesis. Budding yeast is highly polarized during the cell cycle and divides asymmetrically, producing two cells with distinct sizes and fates. Many components of the machinery establishing cell polarization during budding are relocalized to the division site (i.e., the bud neck) for cytokinesis. In this review we recapitulate how budding yeast cells undergo polarized processes at the bud neck for cell division. PMID:27085703

  11. Asymmetric biocatalysis with microbial enzymes and cells.

    PubMed

    Wohlgemuth, Roland

    2010-06-01

    Microbial enzymes and cells continue to be important tools and nature's privileged chiral catalysts for performing asymmetric biocatalysis from the analytical small scale to the preparative and large scale in synthesis and degradation. The application of biocatalysts for preparing molecular asymmetry has achieved high efficiency, enantioselectivity and yield and is experiencing today a worldwide renaissance. Recent developments in the discovery, development and production of stable biocatalysts, in the design of new biocatalytic processes and in the product recovery and purification processes have made biocatalytic approaches using microbial cells and enzymes attractive choices for the synthesis of chiral compounds. The methodologies of kinetic resolution and kinetic asymmetric transformation, dynamic kinetic resolution and deracemization, desymmetrization, asymmetric synthesis with or without diastereo control and multi-step asymmetric biocatalysis are finding increasing applications in research. The ever-increasing use of hydrolytic enzymes has been accompanied by new applications of oxidoreductases, transferases and lyases. Isomerases, already used in large-scale processes, and ligases, are emerging as interesting biocatalysts for new synthetic applications. The production of a wide variety of industrial products by asymmetric biocatalysis has even become the preferred method of production. PMID:20434391

  12. Improved microbial fuel cell performance by encapsulating microbial cells with a nickel-coated sponge.

    PubMed

    Liu, Xueying; Du, Xiaoyu; Wang, Xia; Li, Naiqiang; Xu, Ping; Ding, Yi

    2013-03-15

    Development of novel anodic materials that could facilitate microbial biofilm formation, substrate transfer, and electron transfer is vital to enhance the performance of microbial fuel cells (MFCs). In this work, nickel-coated sponge, as a novel and inexpensive material with an open three-dimensional macro-porous structure, was employed as an anode to encapsulate microbial cells. Compared with planar carbon paper, the nickel-coated sponge did not only offer a high surface area to facilitate microbial cells attachment and colonization but also supported sufficient substrate transfer and electron transfer due to multiplexed and highly conductive pathways. As expected, the resulting nickel-coated sponge biofilm demonstrated excellent electrochemical activity and power output stability during electricity generation processes. A higher maximum power density of 996 mW m(-2) and a longer, more stable electricity generation period were achieved with the nickel-coated sponge biofilm than previously reported results. Notably, chemical oxygen demand (COD) removal reached 90.3% in the anode chamber, suggesting that the nickel-coated sponge is a highly promising anodic material and an efficient immobilization method for the fabrication of MFCs. PMID:22939511

  13. Fermentation Temperature Modulates Phosphatidylethanolamine and Phosphatidylinositol Levels in the Cell Membrane of Saccharomyces cerevisiae

    PubMed Central

    Henderson, Clark M.; Zeno, Wade F.; Lerno, Larry A.; Longo, Marjorie L.

    2013-01-01

    During alcoholic fermentation, Saccharomyces cerevisiae is exposed to a host of environmental and physiological stresses. Extremes of fermentation temperature have previously been demonstrated to induce fermentation arrest under growth conditions that would otherwise result in complete sugar utilization at “normal” temperatures and nutrient levels. Fermentations were carried out at 15°C, 25°C, and 35°C in a defined high-sugar medium using three Saccharomyces cerevisiae strains with diverse fermentation characteristics. The lipid composition of these strains was analyzed at two fermentation stages, when ethanol levels were low early in stationary phase and in late stationary phase at high ethanol concentrations. Several lipids exhibited dramatic differences in membrane concentration in a temperature-dependent manner. Principal component analysis (PCA) was used as a tool to elucidate correlations between specific lipid species and fermentation temperature for each yeast strain. Fermentations carried out at 35°C exhibited very high concentrations of several phosphatidylinositol species, whereas at 15°C these yeast strains exhibited higher levels of phosphatidylethanolamine and phosphatidylcholine species with medium-chain fatty acids. Furthermore, membrane concentrations of ergosterol were highest in the yeast strain that experienced stuck fermentations at all three temperatures. Fluorescence anisotropy measurements of yeast cell membrane fluidity during fermentation were carried out using the lipophilic fluorophore diphenylhexatriene. These measurements demonstrate that the changes in the lipid composition of these yeast strains across the range of fermentation temperatures used in this study did not significantly affect cell membrane fluidity. However, the results from this study indicate that fermenting S. cerevisiae modulates its membrane lipid composition in a temperature-dependent manner. PMID:23811519

  14. Permeabilization of yeast Saccharomyces cerevisiae cell walls using nanosecond high power electrical pulses

    NASA Astrophysics Data System (ADS)

    Stirke, A.; Zimkus, A.; Balevicius, S.; Stankevic, V.; Ramanaviciene, A.; Ramanavicius, A.; Zurauskiene, N.

    2014-12-01

    The electrical field-induced changes of the yeast Saccharomyces cerevisiae cells permeabilization to tetraphenylphosphonium (TPP+) ions were studied using square-shaped, nanosecond duration high power electrical pulses. It was obtained that pulses having durations ranging from 10 ns to 60 ns, and generating electric field strengths up to 190 kV/cm significantly (up to 65 times) increase the absorption rate of TPP+ ions without any detectible influence on the yeast cell viability. The modelling of the TPP+ absorption process using a second order rate equation demonstrates that depending on the duration of the pulses, yeast cell clusters of different sizes are homogeniously permeabilized. It was concluded, that nanosecond pulse-induced permeabilization can be applied to increase the operational speed of whole cell biosensors.

  15. [Water activity and food stability. I. Effects on viability of Saccharomyces cerevisiae cells (author's transl)].

    PubMed

    Guerzoni, M E; Suzzi, G; Lerici, C R; Bartolini, R; Testa, G

    1976-01-01

    Biological activity of microorganism is related to water activity (aw). In this paper the effect of glicerol as humectant on Saccharomyces cerevisiae viability was considered. The irreversible loss of viability was observed only for values inferior than 0,75. The K+ presence promoted an increasing of cell viability and growth. We have evaluated the changes of the most important components of cell poll; the increasing of glicerol amount of the system induced a drastic fall of aminoacids, purines and K ions content, but it increased the Na ions content. The exposure of cells to increasing glicerol concentrations, caused an aminoacids and purines excretion related to contact time; after a few hours this material was readsorbed by cells. PMID:799835

  16. [Cloning and expression of bacteriophage FMV lysocyme gene in cells of yeasts Saccharomyces cerevisiae and Pichia pastoris].

    PubMed

    Kozlov, D G; Cheperigin, S E; Chestkov, A V; Krylov, V N; Tsygankov, Iu D

    2010-03-01

    Cloning, sequencing, and expression of the gene for soluble lysozyme of bacteriophage FMV from Gram-negative Pseudomonas aeruginosa bacteria were conducted in yeast cells. Comparable efficiency of two lysozyme expression variants (as intracellular or secreted proteins) was estimated in cells of Saccharomyces cerevisiae and Pichia pastoris. Under laboratory conditions, yeast S. cerevisiae proved to be more effective producer of phage lysozyme than P. pastoris, the yield of the enzyme in the secreted form being significantly higher than that produced in the intracellular form. PMID:20391778

  17. [Strategies for regulating multiple genes in microbial cell factories].

    PubMed

    Jiang, Tianyi; Li, Lixiang; Ma, Cuiqing; Xu, Ping

    2010-10-01

    Microbial metabolic engineering and synthetic biology are important disciplines of microbial technology nowadays. Microbial cells are fast growing, easy to be cultivated in large scale, clear in genetic background and convenient in genetic modification. They play an important role in many domains. Microbial cell factory means an artificial microbial metabolic system that can be used in chemical production. The construction of a microbial cell factory needs transferring of multiple genes or a whole metabolic pathway, which may cause some problems such as metabolism imbalance and accumulation of mesostates. This review focuses on the regulation strategies of different levels involving simultaneous engagement of multiple genes. Future perspectives on the development of this domain were also discussed. PMID:21218630

  18. A thermophilic microbial fuel cell design

    NASA Astrophysics Data System (ADS)

    Carver, Sarah M.; Vuoriranta, Pertti; Tuovinen, Olli H.

    Microbial fuel cells (MFCs) are reactors able to generate electricity by capturing electrons from the anaerobic respiratory processes of microorganisms. While the majority of MFCs have been tested at ambient or mesophilic temperatures, thermophilic systems warrant evaluation because of the potential for increased microbial activity rates on the anode. MFC studies at elevated temperatures have been scattered, using designs that are already established, specifically air-cathode single chambers and two-chamber designs. This study was prompted by our previous attempts that showed an increased amount of evaporation in thermophilic MFCs, adding unnecessary technical difficulties and causing excessive maintenance. In this paper, we describe a thermophilic MFC design that prevents evaporation. The design was tested at 57 °C with an anaerobic, thermophilic consortium that respired with glucose to generate a power density of 375 mW m -2 after 590 h. Polarization and voltage data showed that the design works in the batch mode but the design allows for adoption to continuous operation.

  19. Saccharomyces Cerevisiae Cell Wall Components as Tools for Ochratoxin A Decontamination

    PubMed Central

    Piotrowska, Małgorzata; Masek, Anna

    2015-01-01

    The aim of this study was to evaluate the usefulness of Saccharomyces cerevisiae cell wall preparations in the adsorption of ochratoxin A (OTA). The study involved the use of a brewer’s yeast cell wall devoid of protein substances, glucans obtained by water and alkaline extraction, a glucan commercially available as a dietary supplement for animals and, additionally, dried brewer’s yeast for comparison. Fourier Transform Infrared (FTIR) analysis of the obtained preparations showed bands characteristic for glucans in the resulting spectra. The yeast cell wall preparation, water-extracted glucan and the commercial glucan bound the highest amount of ochratoxin A, above 55% of the initial concentration, and the alkaline-extracted glucan adsorbed the lowest amount of this toxin. It has been shown that adsorption is most effective at a close-to-neutral pH, while being considerably limited in alkaline conditions. PMID:25848694

  20. Microbial fuel cells and microbial electrolysis cells for the production of bioelectricity and biomaterials.

    PubMed

    Zhou, Minghua; Yang, Jie; Wang, Hongyu; Jin, Tao; Xu, Dake; Gu, Tingyue

    2013-01-01

    Today's global energy crisis requires a multifaceted solution. Bioenergy is an important part of the solution. The microbial fuel cell (MFC) technology stands out as an attractive potential technology in bioenergy. MFCs can convert energy stored in organic matter directly into bioelectricity. MFCs can also be operated in the electrolysis mode as microbial electrolysis cells to produce bioproducts such as hydrogen and ethanol. Various wastewaters containing low-grade organic carbons that are otherwise unutilized can be used as feed streams for MFCs. Despite major advances in the past decade, further improvements in MFC power output and cost reduction are needed for MFCs to be practical. This paper analysed MFC operating principles using bioenergetics and bioelectrochemistry. Several major issues were explored to improve the MFC performance. An emphasis was placed on the use of catalytic materials for MFC electrodes. Recent advances in the production of various biomaterials using MFCs were also investigated. PMID:24350445

  1. Biosensoric potential of microbial fuel cells.

    PubMed

    Schneider, György; Kovács, Tamás; Rákhely, Gábor; Czeller, Miklós

    2016-08-01

    Recent progress in microbial fuel cell (MFC) technology has highlighted the potential of these devices to be used as biosensors. The advantages of MFC-based biosensors are that they are phenotypic and can function in either assay- or flow-through formats. These features make them appropriate for contiguous on-line monitoring in laboratories and for in-field applications. The selectivity of an MFC biosensor depends on the applied microorganisms in the anodic compartment where electron transfer (ET) between the artificial surface (anode) and bacterium occurs. This process strongly determines the internal resistance of the sensoric system and thus influences signal outcome and response time. Despite their beneficial characteristics, the number of MFC-based biosensoric applications has been limited until now. The aim of this mini-review is to turn attention to the biosensoric potential of MFCs by summarizing ET mechanisms on which recently established and future sensoric devices are based. PMID:27401925

  2. Relation between cell death progression, reactive oxygen species production and mitochondrial membrane potential in fermenting Saccharomyces cerevisiae cells under heat-shock conditions.

    PubMed

    Pyatrikas, Darya V; Fedoseeva, Irina V; Varakina, Nina N; Rusaleva, Tatyana M; Stepanov, Alexei V; Fedyaeva, Anna V; Borovskii, Gennadii B; Rikhvanov, Eugene G

    2015-06-01

    Moderate heat shock increased reactive oxygen species (ROS) production that led to cell death in glucose-grown Saccharomyces cerevisiae cells. Conditions that disturb mitochondrial functions such as treatment by uncouplers and petite mutation were shown to inhibit ROS production and protects cell from thermal death. Hence, mitochondria are responsible for ROS production and play an active role in cell death. An increase in ROS production was accompanied by hyperpolarization of inner mitochondrial membrane. All agents suppressing hyperpolarization also suppressed heat-induced ROS production. It was supposed that generation of ROS under moderate heat shock in glucose-grown S. cerevisiae cells is driven by the mitochondrial membrane potential. PMID:25991811

  3. Microbial Fuel Cells and Microbial Ecology: Applications in Ruminant Health and Production Research

    PubMed Central

    Osterstock, Jason B.; Pinchak, William E.; Ishii, Shun’ichi; Nelson, Karen E.

    2009-01-01

    Microbial fuel cell (MFC) systems employ the catalytic activity of microbes to produce electricity from the oxidation of organic, and in some cases inorganic, substrates. MFC systems have been primarily explored for their use in bioremediation and bioenergy applications; however, these systems also offer a unique strategy for the cultivation of synergistic microbial communities. It has been hypothesized that the mechanism(s) of microbial electron transfer that enable electricity production in MFCs may be a cooperative strategy within mixed microbial consortia that is associated with, or is an alternative to, interspecies hydrogen (H2) transfer. Microbial fermentation processes and methanogenesis in ruminant animals are highly dependent on the consumption and production of H2in the rumen. Given the crucial role that H2 plays in ruminant digestion, it is desirable to understand the microbial relationships that control H2 partial pressures within the rumen; MFCs may serve as unique tools for studying this complex ecological system. Further, MFC systems offer a novel approach to studying biofilms that form under different redox conditions and may be applied to achieve a greater understanding of how microbial biofilms impact animal health. Here, we present a brief summary of the efforts made towards understanding rumen microbial ecology, microbial biofilms related to animal health, and how MFCs may be further applied in ruminant research. PMID:20024685

  4. Physical, functional and structural characterization of the cell wall fractions from baker's yeast Saccharomyces cerevisiae.

    PubMed

    Borchani, Chema; Fonteyn, Fabienne; Jamin, Guilhem; Paquot, Michel; Thonart, Philippe; Blecker, Christophe

    2016-03-01

    The yeast cell wall of Saccharomyces cerevisiae is an important source of β-d-glucan, a glucose homopolymer with many functional, nutritional and human health benefits. In the present study, the yeast cell wall fractionation process involving enzymatic treatments (savinase and lipolase enzymes) affected most of the physical and functional characteristics of extracted fractions. Thus, the fractionation process showed that β-d-glucan fraction F4 had significantly higher swelling power and fat binding capacity compared to other fractions (F1, F2 and F3). It also exhibited a viscosity of 652.12mPas and a high degree of brightness of extracted β-d-glucan fraction. Moreover, the fractionation process seemed to have an effect on structural and thermal properties of extracted fractions. Overall, results showed that yeast β-d-glucan had good potential for use as a prebiotic ingredient in food, as well as medicinal and pharmaceutical products. PMID:26471666

  5. Combining magnetic sorting of mother cells and fluctuation tests to analyze genome instability during mitotic cell aging in Saccharomyces cerevisiae.

    PubMed

    Patterson, Melissa N; Maxwell, Patrick H

    2014-01-01

    Saccharomyces cerevisiae has been an excellent model system for examining mechanisms and consequences of genome instability. Information gained from this yeast model is relevant to many organisms, including humans, since DNA repair and DNA damage response factors are well conserved across diverse species. However, S. cerevisiae has not yet been used to fully address whether the rate of accumulating mutations changes with increasing replicative (mitotic) age due to technical constraints. For instance, measurements of yeast replicative lifespan through micromanipulation involve very small populations of cells, which prohibit detection of rare mutations. Genetic methods to enrich for mother cells in populations by inducing death of daughter cells have been developed, but population sizes are still limited by the frequency with which random mutations that compromise the selection systems occur. The current protocol takes advantage of magnetic sorting of surface-labeled yeast mother cells to obtain large enough populations of aging mother cells to quantify rare mutations through phenotypic selections. Mutation rates, measured through fluctuation tests, and mutation frequencies are first established for young cells and used to predict the frequency of mutations in mother cells of various replicative ages. Mutation frequencies are then determined for sorted mother cells, and the age of the mother cells is determined using flow cytometry by staining with a fluorescent reagent that detects bud scars formed on their cell surfaces during cell division. Comparison of predicted mutation frequencies based on the number of cell divisions to the frequencies experimentally observed for mother cells of a given replicative age can then identify whether there are age-related changes in the rate of accumulating mutations. Variations of this basic protocol provide the means to investigate the influence of alterations in specific gene functions or specific environmental conditions on

  6. Cell wall-related bionumbers and bioestimates of Saccharomyces cerevisiae and Candida albicans.

    PubMed

    Klis, Frans M; de Koster, Chris G; Brul, Stanley

    2014-01-01

    Bionumbers and bioestimates are valuable tools in biological research. Here we focus on cell wall-related bionumbers and bioestimates of the budding yeast Saccharomyces cerevisiae and the polymorphic, pathogenic fungus Candida albicans. We discuss the linear relationship between cell size and cell ploidy, the correlation between cell size and specific growth rate, the effect of turgor pressure on cell size, and the reason why using fixed cells for measuring cellular dimensions can result in serious underestimation of in vivo values. We further consider the evidence that individual buds and hyphae grow linearly and that exponential growth of the population results from regular formation of new daughter cells and regular hyphal branching. Our calculations show that hyphal growth allows C. albicans to cover much larger distances per unit of time than the yeast mode of growth and that this is accompanied by strongly increased surface expansion rates. We therefore predict that the transcript levels of genes involved in wall formation increase during hyphal growth. Interestingly, wall proteins and polysaccharides seem barely, if at all, subject to turnover and replacement. A general lesson is how strongly most bionumbers and bioestimates depend on environmental conditions and genetic background, thus reemphasizing the importance of well-defined and carefully chosen culture conditions and experimental approaches. Finally, we propose that the numbers and estimates described here offer a solid starting point for similar studies of other cell compartments and other yeast species. PMID:24243791

  7. Identification of Genes Required for Normal Pheromone-Induced Cell Polarization in Saccharomyces Cerevisiae

    PubMed Central

    Chenevert, J.; Valtz, N.; Herskowitz, I.

    1994-01-01

    In response to mating pheromones, cells of the yeast Saccharomyces cerevisiae adopt a polarized ``shmoo'' morphology, in which the cytoskeleton and proteins involved in mating are localized to a cell-surface projection. This polarization is presumed to reflect the oriented morphogenesis that occurs between mating partners to facilitate cell and nuclear fusion. To identify genes involved in pheromone-induced cell polarization, we have isolated mutants defective in mating to an enfeebled partner and studied a subset of these mutants. The 34 mutants of interest are proficient for pheromone production, arrest in response to pheromone, mate to wild-type strains, and exhibit normal cell polarity during vegetative growth. The mutants were divided into classes based on their morphological responses to mating pheromone. One class is unable to localize cell-surface growth in response to mating factor and instead enlarges in a uniform manner. These mutants harbor special alleles of genes required for cell polarization during vegetative growth, BEM1 and CDC24. Another class of mutants forms bilobed, peanut-like shapes when treated with pheromone and defines two genes, PEA1 and PEA2. PEA1 is identical to SPA2. A third class forms normally shaped but tiny shmoos and defines the gene TNY1. A final group of mutants exhibits apparently normal shmoo morphology. The nature of their mating defect is yet to be determined. We discuss the possible roles of these gene products in establishing cell polarity during mating. PMID:8013906

  8. Microbial Community Analysis of a Single Chamber Microbial Fuel Cell Using Potato Wastewater

    SciTech Connect

    Zhen Li; Rishika Haynes; Eugene Sato; Malcolm Shields; Yoshiko Fujita; Chikashi Sato

    2014-04-01

    Microbial fuel cells (MFCs) convert chemical energy to electrical energy via bioelectrochemical reactions mediated by microorganisms. We investigated the diversity of the microbial community in an air cathode single chamber MFC that utilized potato-process wastewater as substrate. Terminal Restriction Fragment Length Polymorphism (T-RFLP) results indicated that the bacterial communities on the anode, cathode, control electrode, and MFC bulk fluid were similar, but differed dramatically from that of the anaerobic domestic sludge and potato wastewater inoculum. The 16S rDNA sequencing results showed that microbial species detected on the anode were predominantly within the phyla of Proteobacteria, Firmicutes, and Bacteroidetes. Fluorescent microscopy results indicated that there was a clear enhancement of biofilm formation on the anode. Results of this study could help improve understanding of the complexity of microbial communities and optimize the microbial composition for generating electricity by MFCs that utilize potato wastewater.

  9. Reduction of volatile acidity of acidic wines by immobilized Saccharomyces cerevisiae cells.

    PubMed

    Vilela, A; Schuller, D; Mendes-Faia, A; Côrte-Real, M

    2013-06-01

    Excessive volatile acidity in wines is a major problem and is still prevalent because available solutions are nevertheless unsatisfactory, namely, blending the filter-sterilized acidic wine with other wines of lower volatile acidity or using reverse osmosis. We have previously explored the use of an empirical biological deacidification procedure to lower the acetic acid content of wines. This winemaker's enological practice, which consists in refermentation associated with acetic acid consumption by yeasts, is performed by mixing the acidic wine with freshly crushed grapes, musts, or marc from a finished wine fermentation. We have shown that the commercial strain Saccharomyces cerevisiae S26 is able to decrease the volatile acidity of acidic wines with a volatile acidity higher than 1.44 g L(-1) acetic acid, with no detrimental impact on wine aroma. In this study, we aimed to optimize the immobilization of S26 cells in alginate beads for the bioreduction of volatile acidity of acidic wines. We found that S26 cells immobilized in double-layer alginate-chitosan beads could reduce the volatile acidity of an acidic wine (1.1 g L(-1) acetic acid, 12.5 % (v/v) ethanol, pH 3.12) by 28 and 62 % within 72 and 168 h, respectively, associated with a slight decrease in ethanol concentration (0.7 %). Similar volatile acidity removal efficiencies were obtained in medium with high glucose concentration (20 % w/v), indicating that this process may also be useful in the deacidification of grape musts. We, therefore, show that immobilized S. cerevisiae S26 cells in double-layer beads are an efficient alternative to improve the quality of wines with excessive volatile acidity. PMID:23361840

  10. Cellular and molecular engineering of yeast Saccharomyces cerevisiae for advanced biobutanol production.

    PubMed

    Kuroda, Kouichi; Ueda, Mitsuyoshi

    2016-02-01

    Butanol is an attractive alternative energy fuel owing to several advantages over ethanol. Among the microbial hosts for biobutanol production, yeast Saccharomyces cerevisiae has a great potential as a microbial host due to its powerful genetic tools, a history of successful industrial use, and its inherent tolerance to higher alcohols. Butanol production by S. cerevisiae was first attempted by transferring the 1-butanol-producing metabolic pathway from native microorganisms or using the endogenous Ehrlich pathway for isobutanol synthesis. Utilizing alternative enzymes with higher activity, eliminating competitive pathways, and maintaining cofactor balance achieved significant improvements in butanol production. Meeting future challenges, such as enhancing butanol tolerance and implementing a comprehensive strategy by high-throughput screening, would further elevate the biobutanol-producing ability of S. cerevisiae toward an ideal microbial cell factory exhibiting high productivity of biobutanol. PMID:26712533

  11. Chemical and enzymatic extraction of heavy metal binding polymers from isolated cell walls of Saccharomyces cerevisiae

    SciTech Connect

    Brady, D.; Stoll, A.D.; Starke, L.; Duncan, J.R. . Dept. of Biochemistry and Microbiology)

    1994-07-01

    Isolated cell walls of the yeast Saccharomyces cerevisiae were treated by either chemical (alkali and acid) or enzymatic (protease, mannanase or [beta]-glucuronidase) processes to yield partially purified products. These products were partially characterized by infrared analysis. They were subsequently reacted with heavy metal cation solutions and the quantity of metal accumulated by the cell wall material determined. The Cu[sup 2+] ion (0.24, 0.36, 1.12, and 0.60 [mu]mol/mg) was accumulated to a greater extent than either Co[sup 2+] (0.13, 0.32, 0.43, and 0.32 [mu]mol/mg) or Cd[sup 2+] (0.17, 0.34, 0.39, and 0.46 [mu]mol/mg) by yeast cell walls, glucan, mannan, and chitin, respectively. The isolated components each accumulated greater quantities of the cations than the intact cell wall. Removal of the protein component of the yeast cell wall by Pronase caused a 29.5% decrease in metal accumulation by yeast cell walls per mass, indicating that protein is a heavy metal accumulating component. The data indicate that the outer mannan-protein layer of the yeast cell wall is more important than the inner glucan-chitin layer in heavy metal cation accumulation.

  12. A pathway for cell wall anchorage of Saccharomyces cerevisiae alpha-agglutinin.

    PubMed Central

    Lu, C F; Kurjan, J; Lipke, P N

    1994-01-01

    Saccharomyces cerevisiae alpha-agglutinin is a cell wall-anchored adhesion glycoprotein. The previously identified 140-kDa form, which contains a glycosyl-phosphatidylinositol (GPI) anchor (D. Wojciechowicz, C.-F. Lu, J. Kurjan, and P. N. Lipke, Mol. Cell. Biol. 13:2554-2563, 1993), and additional forms of 80, 150, 250 to 300, and > 300 kDa had the properties of intermediates in a transport and cell wall anchorage pathway. N glycosylation and additional modifications resulted in successive increases in size during transport. The 150- and 250- to 300-kDa forms were membrane associated and are likely to be intermediates between the 140-kDa form and a cell surface GPI-anchored form of > 300 kDa. A soluble form of > 300 kDa that lacked the GPI anchor had properties of a periplasmic intermediate between the plasma membrane form and the > 300-kDa cell wall-anchored form. These results constitute experimental support for the hypothesis that GPI anchors act to localize alpha-agglutinin to the plasma membrane and that cell wall anchorage involves release from the GPI anchor to produce a periplasmic intermediate followed by linkage to the cell wall. Images PMID:8007981

  13. Predictive Potential of Flux Balance Analysis of Saccharomyces cerevisiae Using as Optimization Function Combinations of Cell Compartmental Objectives

    PubMed Central

    García Sánchez, Carlos Eduardo; Vargas García, César Augusto; Torres Sáez, Rodrigo Gonzalo

    2012-01-01

    Background The main objective of flux balance analysis (FBA) is to obtain quantitative predictions of metabolic fluxes of an organism, and it is necessary to use an appropriate objective function to guarantee a good estimation of those fluxes. Methodology In this study, the predictive performance of FBA was evaluated, using objective functions arising from the linear combination of different cellular objectives. This approach is most suitable for eukaryotic cells, owing to their multiplicity of cellular compartments. For this reason, Saccharomyces cerevisiae was used as model organism, and its metabolic network was represented using the genome-scale metabolic model iMM904. As the objective was to evaluate the predictive performance from the FBA using the kind of objective function previously described, substrate uptake and oxygen consumption were the only input data used for the FBA. Experimental information about microbial growth and exchange of metabolites with the environment was used to assess the quality of the predictions. Conclusions The quality of the predictions obtained with the FBA depends greatly on the knowledge of the oxygen uptake rate. For the most of studied classifications, the best predictions were obtained with “maximization of growth”, and with some combinations that include this objective. However, in the case of exponential growth with unknown oxygen exchange flux, the objective function “maximization of growth, plus minimization of NADH production in cytosol, plus minimization of NAD(P)H consumption in mitochondrion” gave much more accurate estimations of fluxes than the obtained with any other objective function explored in this study. PMID:22912775

  14. Anethole induces apoptotic cell death accompanied by reactive oxygen species production and DNA fragmentation in Aspergillus fumigatus and Saccharomyces cerevisiae.

    PubMed

    Fujita, Ken-Ichi; Tatsumi, Miki; Ogita, Akira; Kubo, Isao; Tanaka, Toshio

    2014-02-01

    trans-Anethole (anethole), a major component of anise oil, has a broad antimicrobial spectrum, and antimicrobial activity that is weaker than that of other antibiotics on the market. When combined with polygodial, nagilactone E, and n-dodecanol, anethole has been shown to possess significant synergistic antifungal activity against a budding yeast, Saccharomyces cerevisiae, and a human opportunistic pathogenic yeast, Candida albicans. However, the antifungal mechanism of anethole has not been completely determined. We found that anethole stimulated cell death of a human opportunistic pathogenic fungus, Aspergillus fumigatus, in addition to S. cerevisiae. The anethole-induced cell death was accompanied by reactive oxygen species production, metacaspase activation, and DNA fragmentation. Several mutants of S. cerevisiae, in which genes related to the apoptosis-initiating execution signals from mitochondria were deleted, were resistant to anethole. These results suggest that anethole-induced cell death could be explained by oxidative stress-dependent apoptosis via typical mitochondrial death cascades in fungi, including A. fumigatus and S. cerevisiae. PMID:24393541

  15. Power overshoot in two-chambered microbial fuel cell (MFC).

    PubMed

    Nien, Po-Chin; Lee, Chin-Yu; Ho, Kuo-Chuan; Adav, Sunil S; Liu, Lihong; Wang, Aijie; Ren, Nanqi; Lee, Duu-Jong

    2011-04-01

    A two-chamber microbial fuel cell was started using iron-reducing strains as inoculum and acetate as carbon sources. The tested microbial fuel cell had an open-circuit voltage of 0.67 V, and reached 1045 mA m(-2) and a power density of 486 mW m(-2) at 0.46 V before power overshoot occurred. Anodic reactions were identified as the rate-determining steps. Stirring the anolyte insignificantly increased cell performance, suggesting a minimal external mass transfer resistance from the anolyte to the anodic biofilm. Data regression analysis indicates that charge transfer resistance at the biofilm-anode junction was negligible. The order of magnitude estimation of electrical conductance indicates that electron transfer resistance had an insignificant effect on microbial fuel cell performance. Resistance in electrogens for substrate utilization is proposed to induce microbial fuel cell power overshoot. PMID:21295969

  16. Transcriptional activation in an improved whole-cell extract from Saccharomyces cerevisiae.

    PubMed Central

    Woontner, M; Wade, P A; Bonner, J; Jaehning, J A

    1991-01-01

    We report an improved in vitro transcription system for Saccharomyces cerevisiae. Small changes in assay and whole-cell extraction procedures increase selective initiation by RNA polymerase II up to 60-fold over previous conditions (M. Woontner and J. A. Jaehning, J. Biol. Chem. 265:8979-8982, 1990), to levels comparable to those obtained with nuclear extracts. We have found that the simultaneous use of distinguishable templates with and without an upstream activation sequence is critical to the measurement of apparent activation. Transcription from any template was very sensitive to the concentrations of template and nontemplate DNA, extract, and activator (GAL4/VP16). Alterations in reaction conditions led to proportionately greater changes from a template lacking an upstream activation sequence; thus, the apparent ratio of activation is largely dependent on the level of basal transcription. Using optimal conditions for activation, we have also demonstrated activation by a bona fide yeast activator, heat shock transcription factor. Images PMID:1875938

  17. Transcriptional activation in an improved whole-cell extract from Saccharomyces cerevisiae.

    PubMed

    Woontner, M; Wade, P A; Bonner, J; Jaehning, J A

    1991-09-01

    We report an improved in vitro transcription system for Saccharomyces cerevisiae. Small changes in assay and whole-cell extraction procedures increase selective initiation by RNA polymerase II up to 60-fold over previous conditions (M. Woontner and J. A. Jaehning, J. Biol. Chem. 265:8979-8982, 1990), to levels comparable to those obtained with nuclear extracts. We have found that the simultaneous use of distinguishable templates with and without an upstream activation sequence is critical to the measurement of apparent activation. Transcription from any template was very sensitive to the concentrations of template and nontemplate DNA, extract, and activator (GAL4/VP16). Alterations in reaction conditions led to proportionately greater changes from a template lacking an upstream activation sequence; thus, the apparent ratio of activation is largely dependent on the level of basal transcription. Using optimal conditions for activation, we have also demonstrated activation by a bona fide yeast activator, heat shock transcription factor. PMID:1875938

  18. The relationship between enzyme activity, cell geometry, and fitness in Saccharomyces cerevisiae.

    PubMed Central

    Weiss, R L; Kukora, J R; Adams, J

    1975-01-01

    The relationship between enzyme activity, cell geometry, and the ploidy levels has been investigated in Saccharomyces cerevisiae. Diploid cells have 1.57 times the volume of haploid cells under nonlimiting growth conditions (minimal medium). However, when diploid cells are grown under conditions of carbon limitation, they have the same volume as haploid cells. Thus, by altering the environmental conditions, cell size can be varied independently of the degree of ploidy. The results indicate that the basic biochemical parameters of the cell are primarily determined by cell geometry rather than ploidy level. RNA content, protein content, and ornithine transcarbamylase (carbamoylphosphate: L-ornithine carbamoyltransferase, EC 2.1.3.3), tryptophan synthetase [L-serine hydro-lyase (adding indole), EC 4.2.1.20], and invertase (alpha-D-glucoside glucohydrolase, Ec 3.2.1.20) activity are related to cell volume, whereas acid phosphatase (orthophosphoric-monoester phosphohydrolase, EC 3.1.3.2) activity, a cell surface enzyme, is related to the surface area of the cells. Fitness is determined by the activity of certain cell surface enzymes, such as acid phosphatase, diploids would be expected to have a lower fitness than haploids because of the lower surface area/volume ratio. However, when fitness is determined by the activity of an internal enzyme, diploids would be expected to have the same fitness as haploids. Results from competition experiments between haploids and diploids are consistent with these predictions. The significance of these results to the evolution of diploidy as the predominant phase of the life cycle of higher plants and animals is discussed. PMID:1093169

  19. Identification of Proteins Whose Synthesis Is Modulated During the Cell Cycle of Saccharomyces cerevisiae

    PubMed Central

    Lörincz, Attila T.; Miller, Mark J.; Xuong, Nguyen-Huu; Geiduschek, E. Peter

    1982-01-01

    We examined the synthesis and turnover of individual proteins in the Saccharomyces cerevisiae cell cycle. Proteins were pulse-labeled with radioactive isotope (35S or 14C) in cells at discrete cycle stages and then resolved on two-dimensional gels and analyzed by a semiautomatic procedure for quantitating gel electropherogram-autoradiographs. The cells were obtained by one of three methods: (i) isolation of synchronous subpopulations of growing cells by zonal centrifugation; (ii) fractionation of pulse-labeled steady-state cultures according to cell age; and (iii) synchronization of cells with the mating pheromone, α-factor. In confirmation of previous studies, we found that the histones H4, H2A, and H2B were synthesized almost exclusively in the late G1 and early S phases. In addition, we identified eight proteins whose rates of synthesis were modulated in the cell cycle, and nine proteins (of which five, which may well be related, were unstable, with half-lives of 10 to 15 min) that might be regulated in the cell cycle by periodic synthesis, modification, or degradation. Based on the time of maximal labeling in the cell cycle and on experiments with α-factor and hydroxyurea, we assigned the cell cycle proteins to two classes: proteins in class I were labeled principally in early G1 phase and at a late stage of the cycle, whereas those in class II were primarily synthesized at times ranging from late G1 to mid S phase. At least one major control point for the cell cycle proteins occurred between “start” and early S phase. A set of stress-responsive proteins was also identified and analyzed. The rates of synthesis of these proteins were affected by certain perturbations that resulted during selection of synchronous cell populations and by heat shock. Images PMID:14582195

  20. N-hypermannose glycosylation disruption enhances recombinant protein production by regulating secretory pathway and cell wall integrity in Saccharomyces cerevisiae.

    PubMed

    Tang, Hongting; Wang, Shenghuan; Wang, Jiajing; Song, Meihui; Xu, Mengyang; Zhang, Mengying; Shen, Yu; Hou, Jin; Bao, Xiaoming

    2016-01-01

    Saccharomyces cerevisiae is a robust host for heterologous protein expression. The efficient expression of cellulases in S. cerevisiae is important for the consolidated bioprocess that directly converts lignocellulose into valuable products. However, heterologous proteins are often N-hyperglycosylated in S. cerevisiae, which may affect protein activity. In this study, the expression of three heterologous proteins, β-glucosidase, endoglucanase and cellobiohydrolase, was found to be N-hyperglycosylated in S. cerevisiae. To block the formation of hypermannose glycan, these proteins were expressed in strains with deletions in key Golgi mannosyltransferases (Och1p, Mnn9p and Mnn1p), respectively. Their extracellular activities improved markedly in the OCH1 and MNN9 deletion strains. Interestingly, truncation of the N-hypermannose glycan did not increase the specific activity of these proteins, but improved the secretion yield. Further analysis showed OCH1 and MNN9 deletion up-regulated genes in the secretory pathway, such as protein folding and vesicular trafficking, but did not induce the unfolded protein response. The cell wall integrity was also affected by OCH1 and MNN9 deletion, which contributed to the release of secretory protein extracellularly. This study demonstrated that mannosyltransferases disruption improved protein secretion through up-regulating secretory pathway and affecting cell wall integrity and provided new insights into glycosylation engineering for protein secretion. PMID:27156860

  1. N-hypermannose glycosylation disruption enhances recombinant protein production by regulating secretory pathway and cell wall integrity in Saccharomyces cerevisiae

    PubMed Central

    Tang, Hongting; Wang, Shenghuan; Wang, Jiajing; Song, Meihui; Xu, Mengyang; Zhang, Mengying; Shen, Yu; Hou, Jin; Bao, Xiaoming

    2016-01-01

    Saccharomyces cerevisiae is a robust host for heterologous protein expression. The efficient expression of cellulases in S. cerevisiae is important for the consolidated bioprocess that directly converts lignocellulose into valuable products. However, heterologous proteins are often N-hyperglycosylated in S. cerevisiae, which may affect protein activity. In this study, the expression of three heterologous proteins, β-glucosidase, endoglucanase and cellobiohydrolase, was found to be N-hyperglycosylated in S. cerevisiae. To block the formation of hypermannose glycan, these proteins were expressed in strains with deletions in key Golgi mannosyltransferases (Och1p, Mnn9p and Mnn1p), respectively. Their extracellular activities improved markedly in the OCH1 and MNN9 deletion strains. Interestingly, truncation of the N-hypermannose glycan did not increase the specific activity of these proteins, but improved the secretion yield. Further analysis showed OCH1 and MNN9 deletion up-regulated genes in the secretory pathway, such as protein folding and vesicular trafficking, but did not induce the unfolded protein response. The cell wall integrity was also affected by OCH1 and MNN9 deletion, which contributed to the release of secretory protein extracellularly. This study demonstrated that mannosyltransferases disruption improved protein secretion through up-regulating secretory pathway and affecting cell wall integrity and provided new insights into glycosylation engineering for protein secretion. PMID:27156860

  2. Microscale microbial fuel cells: Advances and challenges.

    PubMed

    Choi, Seokheun

    2015-07-15

    The next generation of sustainable energy could come from microorganisms; evidence that it can be seen with the given rise of Electromicrobiology, the study of microorganisms' electrical properties. Many recent advances in electromicrobiology stem from studying microbial fuel cells (MFCs), which are gaining acceptance as a future alternative "green" energy technology and energy-efficient wastewater treatment method. MFCs are powered by living microorganisms with clean and sustainable features; they efficiently catalyse the degradation of a broad range of organic substrates under natural conditions. There is also increasing interest in photosynthetic MFCs designed to harness Earth's most abundant and promising energy source (solar irradiation). Despite their vast potential and promise, however, MFCs and photosynthetic MFCs have not yet successfully translated into commercial applications because they demonstrate persistent performance limitations and bottlenecks associated with scaling up. Instead, microscale MFCs have received increasing attention as a unique platform for various applications such as powering small portable electronic elements in remote locations, performing fundamental studies of microorganisms, screening bacterial strains, and toxicity detection in water. Furthermore, the stacking of miniaturized MFCs has been demonstrated to offer larger power densities than a single macroscale MFC in terms of scaling up. In this overview, we discuss recent achievements in microscale MFCs as well as their potential applications. Further scientific and technological challenges are also reviewed. PMID:25703724

  3. AC power generation from microbial fuel cells

    NASA Astrophysics Data System (ADS)

    Lobo, Fernanda Leite; Wang, Heming; Forrestal, Casey; Ren, Zhiyong Jason

    2015-11-01

    Microbial fuel cells (MFCs) directly convert biodegradable substrates to electricity and carry good potential for energy-positive wastewater treatment. However, the low and direct current (DC) output from MFC is not usable for general electronics except small sensors, yet commercial DC-AC converters or inverters used in solar systems cannot be directly applied to MFCs. This study presents a new DC-AC converter system for MFCs that can generate alternating voltage in any desired frequency. Results show that AC power can be easily achieved in three different frequencies tested (1, 10, 60 Hz), and no energy storage layer such as capacitors was needed. The DC-AC converter efficiency was higher than 95% when powered by either individual MFCs or simple MFC stacks. Total harmonic distortion (THD) was used to investigate the quality of the energy, and it showed that the energy could be directly usable for linear electronic loads. This study shows that through electrical conversion MFCs can be potentially used in household electronics for decentralized off-grid communities.

  4. Potentiation of gene targeting in human cells by expression of Saccharomyces cerevisiae Rad52.

    PubMed

    Di Primio, Cristina; Galli, Alvaro; Cervelli, Tiziana; Zoppè, Monica; Rainaldi, Giuseppe

    2005-01-01

    When exogenous DNA is stably introduced in mammalian cells, it is typically integrated in random positions, and only a minor fraction enters a pathway of homologous recombination (HR). The complex Rad51/Rad52 is a major player in the management of exogenous DNA in eukaryotic organisms and plays a critical role in the choice of repair system. In Saccharomyces cerevisiae, the pathway of choice is HR, mediated by Rad52 (ScRad52), which differs slightly from its human homologue. Here, we present an approach that utilizes ScRad52 to enhance HR in human cells containing a specific substrate for recombination. Clones of HeLa cells were produced expressing functional ScRad52. These cells showed enhanced resistance to DNA damaging treatments and revealed a different distribution of Rad51 foci (a marker of recombination complex formation). More significantly, ScRad52 expression resulted in an up to 37-fold increase in gene targeting by HR. In the same cells, random integration of exogenous DNA was significantly reduced, consistent with the view that HR and non-homologous end joining are alternative competing pathways. Expression of ScRad52 could offer a major improvement for experiments requiring gene targeting by HR, both in basic research and in gene therapy studies. PMID:16106043

  5. Potentiation of gene targeting in human cells by expression of Saccharomyces cerevisiae Rad52

    PubMed Central

    Di Primio, Cristina; Galli, Alvaro; Cervelli, Tiziana; Zoppè, Monica; Rainaldi, Giuseppe

    2005-01-01

    When exogenous DNA is stably introduced in mammalian cells, it is typically integrated in random positions, and only a minor fraction enters a pathway of homologous recombination (HR). The complex Rad51/Rad52 is a major player in the management of exogenous DNA in eukaryotic organisms and plays a critical role in the choice of repair system. In Saccharomyces cerevisiae, the pathway of choice is HR, mediated by Rad52 (ScRad52), which differs slightly from its human homologue. Here, we present an approach that utilizes ScRad52 to enhance HR in human cells containing a specific substrate for recombination. Clones of HeLa cells were produced expressing functional ScRad52. These cells showed enhanced resistance to DNA damaging treatments and revealed a different distribution of Rad51 foci (a marker of recombination complex formation). More significantly, ScRad52 expression resulted in an up to 37-fold increase in gene targeting by HR. In the same cells, random integration of exogenous DNA was significantly reduced, consistent with the view that HR and non-homologous end joining are alternative competing pathways. Expression of ScRad52 could offer a major improvement for experiments requiring gene targeting by HR, both in basic research and in gene therapy studies. PMID:16106043

  6. [Progress in nanomaterials modified anodes of microbial fuel cell].

    PubMed

    Niu, Hao; Wu, Wenguo

    2016-03-01

    Anode is an important part of microbial fuel cell, its performance significantly affects the electricity generation of microbial fuel cells (MFCs). Nanomaterials have excellent properties, such as good conductivity and large surface area. Therefore, nanomaterials modified anode can effectively reduce the electrode resistance, increase the amount of microbial adhesion and improve the electricity generation of MFCs. In this paper, we introduced various nanomaterials modified anodes and summarized their effects on the output performance of MFCs. Finally, the prospect of modifying nanomaterials and technologies were discussed. PMID:27349110

  7. Implementation of microbial fuel cell in harvesting energy using wastewater

    NASA Astrophysics Data System (ADS)

    Ramli, N. L.; Wahab, M. S. Abdul; Sharif, S. A. Md; Ramly, N. H.

    2016-02-01

    In this century, most of the companies use the electricity from the fossils fuels such as oil, gas and coal. This method will give negative impact to the environment and the fossils fuel will be run out. This project is to develop a microbial fuels cell that can produce electricity. There are several types of the microbial fuel cell, which are a single chamber, double chamber and continuous. In this paper, the double chamber microbial fuel cell was selected to investigate the effect of suspended sludge into the double chamber microbial fuels cell. The salt bridge will construct between both chambers of the double chamber microbial fuels cell. Carbon graphite rod is selected as an electrode at the cathode and anode to transfer the electron from the anode to the cathode. Electricity is generated from the anaerobic oxidation of organic matter by bacteria. At the end of this project, the microbial fuels cell was successful in generating electricity that can be used for a specific application.

  8. Identification of a mannoprotein present in the inner layer of the cell wall of Saccharomyces cerevisiae.

    PubMed Central

    Moukadiri, I; Armero, J; Abad, A; Sentandreu, R; Zueco, J

    1997-01-01

    Cell wall extracts from the double-mutant mnn1 mnn9 strain were used as the immunogen to obtain a monoclonal antibody (MAb), SAC A6, that recognizes a specific mannoprotein--which we have named Icwp--in the walls of cells of Saccharomyces cerevisiae. Icwp runs as a polydisperse band of over 180 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of Zymolyase extracts of cell walls, although an analysis of the secretory pattern of the mannoprotein shows that at the level of secretory vesicles, it behaves like a discrete band of 140 kDa. Immunofluorescence analysis with the MAb showed that Icwp lies at the inner layer of the cell wall, being accessible to the antibody only after the outer layer of mannoproteins is disturbed by treatment with tunicamycin. The screening of a lambda gt11 expression library enabled us to identify the open reading frame (ORF) coding for Icwp. ICWP (EMBL accession number YLR391w, frame +3) codes for 238 amino acids, of which over 40% are serine or threonine, and contains a putative N-glycosylation site and a putative glycosylphosphatidylinositol attachment signal. Both disruption and overexpression of the ORF caused increased sensitivities to calcofluor white and Congo red, while the disruption caused an increased sensitivity to Zymolyase digestion, suggesting for Icwp a structural role in association with glucan. PMID:9079899

  9. Septin-Dependent Assembly of a Cell Cycle-Regulatory Module in Saccharomyces cerevisiae

    PubMed Central

    Longtine, Mark S.; Theesfeld, Chandra L.; McMillan, John N.; Weaver, Elizabeth; Pringle, John R.; Lew, Daniel J.

    2000-01-01

    Saccharomyces cerevisiae septin mutants have pleiotropic defects, which include the formation of abnormally elongated buds. This bud morphology results at least in part from a cell cycle delay imposed by the Cdc28p-inhibitory kinase Swe1p. Mutations in three other genes (GIN4, encoding a kinase related to the Schizosaccharomyces pombe mitotic inducer Nim1p; CLA4, encoding a p21-activated kinase; and NAP1, encoding a Clb2p-interacting protein) also produce perturbations of septin organization associated with an Swe1p-dependent cell cycle delay. The effects of gin4, cla4, and nap1 mutations are additive, indicating that these proteins promote normal septin organization through pathways that are at least partially independent. In contrast, mutations affecting the other two Nim1p-related kinases in S. cerevisiae, Hsl1p and Kcc4p, produce no detectable effect on septin organization. However, deletion of HSL1, but not of KCC4, did produce a cell cycle delay under some conditions; this delay appears to reflect a direct role of Hsl1p in the regulation of Swe1p. As shown previously, Swe1p plays a central role in the morphogenesis checkpoint that delays the cell cycle in response to defects in bud formation. Swe1p is localized to the nucleus and to the daughter side of the mother bud neck prior to its degradation in G2/M phase. Both the neck localization of Swe1p and its degradation require Hsl1p and its binding partner Hsl7p, both of which colocalize with Swe1p at the daughter side of the neck. This localization is lost in mutants with perturbed septin organization, suggesting that the release of Hsl1p and Hsl7p from the neck may reduce their ability to inactivate Swe1p and thus contribute to the G2 delay observed in such mutants. In contrast, treatments that perturb actin organization have little effect on Hsl1p and Hsl7p localization, suggesting that such treatments must stabilize Swe1p by another mechanism. The apparent dependence of Swe1p degradation on localization of

  10. An atypical active cell death process underlies the fungicidal activity of ciclopirox olamine against the yeast Saccharomyces cerevisiae.

    PubMed

    Almeida, Bruno; Sampaio-Marques, Belém; Carvalho, Joana; Silva, Manuel T; Leão, Cecília; Rodrigues, Fernando; Ludovico, Paula

    2007-05-01

    Ciclopirox olamine (CPO), a fungicidal agent widely used in clinical practice, induced in Saccharomyces cerevisiae an active cell death (ACD) process characterized by changes in nuclear morphology and chromatin condensation associated with the appearance of a population in the sub-G(0)/G(1) cell cycle phase and an arrest delay in the G(2)/M phases. This ACD was associated neither with intracellular reactive oxygen species (ROS) signaling, as revealed by the use of different classes of ROS scavengers, nor with a terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-positive phenotype. Furthermore, CPO-induced cell death seems to be dependent on unknown protease activity but independent of the apoptotic regulators Aif1p and Yca1p and of autophagic pathways involving Apg5p, Apg8p and Uth1p. Our results show that CPO triggers in S. cerevisiae an atypical nonapoptotic, nonautophagic ACD with as yet unknown regulators. PMID:17233764

  11. Microbial corrosion monitoring by an amperometric microbial biosensor developed using whole cell of Pseudomonas sp.

    PubMed

    Dubey, R S; Upadhyay, S N

    2001-12-01

    A microbial biosensor was developed for monitoring microbiologically influenced corrosion (MIC) of metallic materials in industrial systems. The Pseudomonas sp. isolated from corroded metal surface was immobilized on acetylcellulose membrane and its respiratory activity was estimated by measuring oxygen consumption. The microbial biosensor was used for the measurement of sulfuric acid in a batch culture medium contaminated by microorganisms. A linear relationship between the microbial sensor response and the concentration of sulfuric acid was observed. The response time of biosensor was 5 min and was dependent on the immobilized cell loading of Pseudomonas sp., pH, temperature and corrosive environments. The microbial biosensor response was stable, reproducible and specific for sensing of sulfur oxidizing bacterial activity. PMID:11679280

  12. Rapid detection of microbial cell abundance in aquatic systems.

    PubMed

    Rocha, Andrea M; Yuan, Quan; Close, Dan M; O'Dell, Kaela B; Fortney, Julian L; Wu, Jayne; Hazen, Terry C

    2016-11-15

    The detection and quantification of naturally occurring microbial cellular densities is an essential component of environmental systems monitoring. While there are a number of commonly utilized approaches for monitoring microbial abundance, capacitance-based biosensors represent a promising approach because of their low-cost and label-free detection of microbial cells, but are not as well characterized as more traditional methods. Here, we investigate the applicability of enhanced alternating current electrokinetics (ACEK) capacitive sensing as a new application for rapidly detecting and quantifying microbial cellular densities in cultured and environmentally sourced aquatic samples. ACEK capacitive sensor performance was evaluated using two distinct and dynamic systems - the Great Australian Bight and groundwater from the Oak Ridge Reservation in Oak Ridge, TN. Results demonstrate that ACEK capacitance-based sensing can accurately determine microbial cell counts throughout cellular concentrations typically encountered in naturally occurring microbial communities (10(3)-10(6) cells/mL). A linear relationship was observed between cellular density and capacitance change correlations, allowing a simple linear curve fitting equation to be used for determining microbial abundances in unknown samples. This work provides a foundation for understanding the limits of capacitance-based sensing in natural environmental samples and supports future efforts focusing on evaluating the robustness ACEK capacitance-based within aquatic environments. PMID:27315516

  13. An efficient magnetically modified microbial cell biocomposite for carbazole biodegradation

    NASA Astrophysics Data System (ADS)

    Li, Yufei; Du, Xiaoyu; Wu, Chao; Liu, Xueying; Wang, Xia; Xu, Ping

    2013-12-01

    Magnetic modification of microbial cells enables to prepare smart biocomposites in bioremediation. In this study, we constructed an efficient biocomposite by assembling Fe3O4 nanoparticles onto the surface of Sphingomonas sp. XLDN2-5 cells. The average particle size of Fe3O4 nanoparticles was about 20 nm with 45.5 emu g-1 saturation magnetization. The morphology of Sphingomonas sp. XLDN2-5 cells before and after Fe3O4 nanoparticle loading was verified by scanning electron microscopy and transmission electronic microscopy. Compared with free cells, the microbial cell/Fe3O4 biocomposite had the same biodegradation activity but exhibited remarkable reusability. The degradation activity of the microbial cell/Fe3O4 biocomposite increased gradually during recycling processes. Additionally, the microbial cell/Fe3O4 biocomposite could be easily separated and recycled by an external magnetic field due to the super-paramagnetic properties of Fe3O4 nanoparticle coating. These results indicated that magnetically modified microbial cells provide a promising technique for improving biocatalysts used in the biodegradation of hazardous compounds.

  14. An efficient magnetically modified microbial cell biocomposite for carbazole biodegradation

    PubMed Central

    2013-01-01

    Magnetic modification of microbial cells enables to prepare smart biocomposites in bioremediation. In this study, we constructed an efficient biocomposite by assembling Fe3O4 nanoparticles onto the surface of Sphingomonas sp. XLDN2-5 cells. The average particle size of Fe3O4 nanoparticles was about 20 nm with 45.5 emu g-1 saturation magnetization. The morphology of Sphingomonas sp. XLDN2-5 cells before and after Fe3O4 nanoparticle loading was verified by scanning electron microscopy and transmission electronic microscopy. Compared with free cells, the microbial cell/Fe3O4 biocomposite had the same biodegradation activity but exhibited remarkable reusability. The degradation activity of the microbial cell/Fe3O4 biocomposite increased gradually during recycling processes. Additionally, the microbial cell/Fe3O4 biocomposite could be easily separated and recycled by an external magnetic field due to the super-paramagnetic properties of Fe3O4 nanoparticle coating. These results indicated that magnetically modified microbial cells provide a promising technique for improving biocatalysts used in the biodegradation of hazardous compounds. PMID:24330511

  15. Polyphosphate is involved in cell cycle progression and genomic stability in Saccharomyces cerevisiae.

    PubMed

    Bru, Samuel; Martínez-Laínez, Joan Marc; Hernández-Ortega, Sara; Quandt, Eva; Torres-Torronteras, Javier; Martí, Ramón; Canadell, David; Ariño, Joaquin; Sharma, Sushma; Jiménez, Javier; Clotet, Josep

    2016-08-01

    Polyphosphate (polyP) is a linear chain of up to hundreds of inorganic phosphate residues that is necessary for many physiological functions in all living organisms. In some bacteria, polyP supplies material to molecules such as DNA, thus playing an important role in biosynthetic processes in prokaryotes. In the present study, we set out to gain further insight into the role of polyP in eukaryotic cells. We observed that polyP amounts are cyclically regulated in Saccharomyces cerevisiae, and those mutants that cannot synthesise (vtc4Δ) or hydrolyse polyP (ppn1Δ, ppx1Δ) present impaired cell cycle progression. Further analysis revealed that polyP mutants show delayed nucleotide production and increased genomic instability. Based on these findings, we concluded that polyP not only maintains intracellular phosphate concentrations in response to fluctuations in extracellular phosphate levels, but also muffles internal cyclic phosphate fluctuations, such as those produced by the sudden demand of phosphate to synthetize deoxynucleotides just before and during DNA duplication. We propose that the presence of polyP in eukaryotic cells is required for the timely and accurate duplication of DNA. PMID:27072996

  16. Saccharomyces cerevisiae STE6 gene product: a novel pathway for protein export in eukaryotic cells.

    PubMed Central

    Kuchler, K; Sterne, R E; Thorner, J

    1989-01-01

    Saccharomyces cerevisiae MATa cells release a lipopeptide mating pheromone, a-factor. Radiolabeling and immunoprecipitation show that MATa ste6 mutants produce pro-a-factor and mature a-factor intracellularly, but little or no extracellular pheromone. Normal MATa cells carrying a multicopy plasmid containing both MFa1 (pro-a-factor structural gene) and the STE6 gene secrete a-factor at least five times faster than the same cells carrying only MFa1 in the same vector. The nucleotide sequence of the STE6 gene predicts a 1290 residue polypeptide with multiple membrane spanning segments and two hydrophilic domains, each strikingly homologous to a set of well-characterized prokaryotic permeases (including hlyB, oppD, hisP, malK and pstB) and sharing even greater identity with mammalian mdr (multiple drug resistance) transporters. These results suggest that the STE6 protein in yeast, and possibly mdr in animals, is a transmembrane translocator that exports polypeptides by a route independent of the classical secretory pathway. Images PMID:2686977

  17. SLA2 mutations cause SWE1-mediated cell cycle phenotypes in Candida albicans and Saccharomyces cerevisiae.

    PubMed

    Gale, Cheryl A; Leonard, Michelle D; Finley, Kenneth R; Christensen, Leah; McClellan, Mark; Abbey, Darren; Kurischko, Cornelia; Bensen, Eric; Tzafrir, Iris; Kauffman, Sarah; Becker, Jeff; Berman, Judith

    2009-12-01

    The early endocytic patch protein Sla2 is important for morphogenesis and growth rates in Saccharomyces cerevisiae and Candida albicans, but the mechanism that connects these processes is not clear. Here we report that growth defects in cells lacking CaSLA2 or ScSLA2 are associated with a cell cycle delay that is influenced by Swe1, a morphogenesis checkpoint kinase. To establish how Swe1 monitors Sla2 function, we compared actin organization and cell cycle dynamics in strains lacking other components of early endocytic patches (Sla1 and Abp1) with those in strains lacking Sla2. Only sla2 strains had defects in actin cables, a known trigger of the morphogenesis checkpoint, yet all three strains exhibited Swe1-dependent phenotypes. Thus, Swe1 appears to monitor actin patch in addition to actin cable function. Furthermore, Swe1 contributed to virulence in a mouse model of disseminated candidiasis, implying a role for the morphogenesis checkpoint during the pathogenesis of C. albicans infections. PMID:19778960

  18. Enrichment of Microbial Electrolysis Cell Biocathodes from Sediment Microbial Fuel Cell Bioanodes

    SciTech Connect

    Pisciotta, JM; Zaybak, Z; Call, DF; Nam, JY; Logan, BE

    2012-07-18

    Electron-accepting (electrotrophic) biocathodes were produced by first enriching graphite fiber brush electrodes as the anodes in sediment-type microbial fuel cells (sMFCs) using two different marine sediments and then electrically inverting the anodes to function as cathodes in two-chamber bioelectrochemical systems (BESs). Electron consumption occurred at set potentials of -439 mV and -539 mV (versus the potential of a standard hydrogen electrode) but not at -339 mV in minimal media lacking organic sources of energy. Results at these different potentials were consistent with separate linear sweep voltammetry (LSV) scans that indicated enhanced activity (current consumption) below only ca. -400 mV. MFC bioanodes not originally acclimated at a set potential produced electron-accepting (electrotrophic) biocathodes, but bioanodes operated at a set potential (+11 mV) did not. CO, was removed from cathode headspace, indicating that the electrotrophic biocathodes were autotrophic. Hydrogen gas generation, followed by loss of hydrogen gas and methane production in one sample, suggested hydrogenotrophic methanogenesis. There was abundant microbial growth in the biocathode chamber, as evidenced by an increase in turbidity and the presence of microorganisms on the cathode surface. Clone library analysis of 16S rRNA genes indicated prominent sequences most similar to those of Eubacterium limosum (Butyribacterium methylotrophicum), Desulfovibrio sp. A2, Rhodococcus opacus, and Gemmata obscuriglobus. Transfer of the suspension to sterile cathodes made of graphite plates, carbon rods, or carbon brushes in new BESs resulted in enhanced current after 4 days, demonstrating growth by these microbial communities on a variety of cathode substrates. This report provides a simple and effective method for enriching autotrophic electrotrophs by the use of sMFCs without the need for set potentials, followed by the use of potentials more negative than -400 mV.

  19. Enrichment of Microbial Electrolysis Cell Biocathodes from Sediment Microbial Fuel Cell Bioanodes

    PubMed Central

    Pisciotta, John M.; Zaybak, Zehra; Call, Douglas F.; Nam, Joo-Youn

    2012-01-01

    Electron-accepting (electrotrophic) biocathodes were produced by first enriching graphite fiber brush electrodes as the anodes in sediment-type microbial fuel cells (sMFCs) using two different marine sediments and then electrically inverting the anodes to function as cathodes in two-chamber bioelectrochemical systems (BESs). Electron consumption occurred at set potentials of −439 mV and −539 mV (versus the potential of a standard hydrogen electrode) but not at −339 mV in minimal media lacking organic sources of energy. Results at these different potentials were consistent with separate linear sweep voltammetry (LSV) scans that indicated enhanced activity (current consumption) below only ca. −400 mV. MFC bioanodes not originally acclimated at a set potential produced electron-accepting (electrotrophic) biocathodes, but bioanodes operated at a set potential (+11 mV) did not. CO2 was removed from cathode headspace, indicating that the electrotrophic biocathodes were autotrophic. Hydrogen gas generation, followed by loss of hydrogen gas and methane production in one sample, suggested hydrogenotrophic methanogenesis. There was abundant microbial growth in the biocathode chamber, as evidenced by an increase in turbidity and the presence of microorganisms on the cathode surface. Clone library analysis of 16S rRNA genes indicated prominent sequences most similar to those of Eubacterium limosum (Butyribacterium methylotrophicum), Desulfovibrio sp. A2, Rhodococcus opacus, and Gemmata obscuriglobus. Transfer of the suspension to sterile cathodes made of graphite plates, carbon rods, or carbon brushes in new BESs resulted in enhanced current after 4 days, demonstrating growth by these microbial communities on a variety of cathode substrates. This report provides a simple and effective method for enriching autotrophic electrotrophs by the use of sMFCs without the need for set potentials, followed by the use of potentials more negative than −400 mV. PMID:22610438

  20. Electricity production from municipal solid waste using microbial fuel cells.

    PubMed

    Chiu, H Y; Pai, T Y; Liu, M H; Chang, C A; Lo, F C; Chang, T C; Lo, H M; Chiang, C F; Chao, K P; Lo, W Y; Lo, S W; Chu, Y L

    2016-07-01

    The organic content of municipal solid waste has long been an attractive source of renewable energy, mainly as a solid fuel in waste-to-energy plants. This study focuses on the potential to use microbial fuel cells to convert municipal solid waste organics into energy using various operational conditions. The results showed that two-chamber microbial fuel cells with carbon felt and carbon felt allocation had a higher maximal power density (20.12 and 30.47 mW m(-2) for 1.5 and 4 L, respectively) than those of other electrode plate allocations. Most two-chamber microbial fuel cells (1.5 and 4 L) had a higher maximal power density than single-chamber ones with corresponding electrode plate allocations. Municipal solid waste with alkali hydrolysis pre-treatment and K3Fe(CN)6 as an electron acceptor improved the maximal power density to 1817.88 mW m(-2) (~0.49% coulomb efficiency, from 0.05-0.49%). The maximal power density from experiments using individual 1.5 and 4 L two-chamber microbial fuel cells, and serial and parallel connections of 1.5 and 4 L two-chamber microbial fuel cells, was found to be in the order of individual 4 L (30.47 mW m(-2)) > serial connection of 1.5 and 4 L (27.75) > individual 1.5 L (20.12) > parallel connection of 1.5 and 4 L (17.04) two-chamber microbial fuel cells . The power density using municipal solid waste microbial fuel cells was compared with information in the literature and discussed. PMID:27231132

  1. Modeling of Sustainable Base Production by Microbial Electrolysis Cell.

    PubMed

    Blatter, Maxime; Sugnaux, Marc; Comninellis, Christos; Nealson, Kenneth; Fischer, Fabian

    2016-07-01

    A predictive model for the microbial/electrochemical base formation from wastewater was established and compared to experimental conditions within a microbial electrolysis cell. A Na2 SO4 /K2 SO4 anolyte showed that model prediction matched experimental results. Using Shewanella oneidensis MR-1, a strong base (pH≈13) was generated using applied voltages between 0.3 and 1.1 V. Due to the use of bicarbonate, the pH value in the anolyte remained unchanged, which is required to maintain microbial activity. PMID:27265318

  2. Removing heavy metals from synthetic effluents using "kamikaze" Saccharomyces cerevisiae cells.

    PubMed

    Ruta, Lavinia; Paraschivescu, Codruta; Matache, Mihaela; Avramescu, Sorin; Farcasanu, Ileana Cornelia

    2010-01-01

    One key step of the bioremediation processes designed to clean up heavy metal contaminated environments is growing resistant cells that accumulate the heavy metals to ensure better removal through a combination of biosorption and continuous metabolic uptake after physical adsorption. Saccharomyces cerevisiae cells can easily act as cation biosorbents, but isolation of mutants that are both hyperaccumulating and tolerant to heavy metals proved extremely difficult. Instead, mutants that are hypersensitive to heavy metals due to increased and continuous uptake from the environment were considered, aiming to use such mutants to reduce the heavy metal content of contaminated waters. In this study, the heavy metal hypersensitive yeast strain pmr1Delta was investigated for the ability to remove Mn2+, Cu2+, Co2+, or Cd2+ from synthetic effluents. Due to increased metal accumulation, the mutant strain was more efficient than the wild-type in removing Mn2+, Cu2+, or Co2+ from synthetic effluents containing 1-2 mM cations, with a selectivity and also in removing Mn2+ and Cd2+ from synthetic effluents containing 20-50 microM cations, with a selectivity Mn2+ > Cd2+. PMID:19795117

  3. Continuous bioethanol production from oilseed rape straw hydrosylate using immobilised Saccharomyces cerevisiae cells.

    PubMed

    Mathew, Anil Kuruvilla; Crook, Mitch; Chaney, Keith; Humphries, Andrea Clare

    2014-02-01

    The aim of the study was to evaluate continuous bioethanol production from oilseed rape (OSR) straw hydrolysate using Saccharomyces cerevisiae cells immobilised in Lentikat® discs. The study evaluated the effect of dilution rate (0.25, 0.50, 0.75 and 1.00 h(-1)), substrate concentration (15, 22, 40 and 60 g L(-1)) and cell loading (0.03, 0.16 and 0.24 g d.c.w.mL(-1) Lentikat®) on bioethanol production. Volumetric productivity was found to increase with increasing substrate concentration from 15 g L(-1) to 60 g L(-1). A maximum volumetric productivity of 12.88 g L(-1)h(-1) was achieved at a substrate concentration of 60 g L(-1) and at a dilution rate of 0.5h(-1). An overall mass balance for bioethanol production was created to determine the energy recovery from bioethanol and concluded that a biorefinery approach might be the most appropriate option for maximising the energy recovery from OSR straw. PMID:24406845

  4. New integrative modules for multicolor-protein labeling and live-cell imaging in Saccharomyces cerevisiae.

    PubMed

    Malcova, Ivana; Farkasovsky, Marian; Senohrabkova, Lenka; Vasicova, Pavla; Hasek, Jiri

    2016-05-01

    Live-imaging analysis is performed in many laboratories all over the world. Various tools have been developed to enable protein labeling either in plasmid or genomic context in live yeast cells. Here, we introduce a set of nine integrative modules for the C-terminal gene tagging that combines three fluorescent proteins (FPs)-ymTagBFP, mCherry and yTagRFP-T with three dominant selection markers: geneticin, nourseothricin and hygromycin. In addition, the construction of two episomal modules for Saccharomyces cerevisiae with photostable yTagRFP-T is also referred to. Our cassettes with orange, red and blue FPs can be combined with other fluorescent probes like green fluorescent protein to prepare double- or triple-labeled strains for multicolor live-cell imaging. Primers for PCR amplification of the cassettes were designed in such a way as to be fully compatible with the existing PCR toolbox representing over 50 various integrative modules and also with deletion cassettes either for single or repeated usage to enable a cost-effective and an easy exchange of tags. New modules can also be used for biochemical analysis since antibodies are available for all three fluorescent probes. PMID:26994102

  5. Efficacy of the direct-fed microbial Enterococcus faecium alone or in combination with Saccharomyces cerevisiae or Lactococcus lactis during induced subacute ruminal acidosis.

    PubMed

    Chiquette, J; Lagrost, J; Girard, C L; Talbot, G; Li, S; Plaizier, J C; Hindrichsen, I K

    2015-01-01

    This study aimed at investigating Enterococcus faecium alone or E. faecium in combination with Saccharomyces cerevisiae or Lactococcus lactis during a subacute ruminal acidosis (SARA) challenge. Four ruminally fistulated Holstein dairy cows were assigned to the following treatments in a 4×4 Latin square design: (1) control (CON); (2) E. faecium (EF); (3) EF + S. cerevisiae (EFSC); (4) EF + L. lactis DSM 11037 (EFLL). Each experimental period consisted of 18 d of adaptation to the respective direct-fed microbial, 3 d of SARA challenge, and 7d of rest. Rumen pH was recorded every 10 min over 24 h on d 17 of adaptation, d 2 of SARA, and d 6 of rest. On the last day of adaptation, SARA, and rest, samples of rumen content (0 and 3 h after feeding) were taken for volatile fatty acids, lactate, vitamin B12, rumen microbes, and lipopolysaccharides determination. Blood samples (0 and 6 h after feeding) were taken for the measurement of acute-phase proteins. Dry matter intake and milk yield were recorded daily. During SARA, mean rumen pH with EFSC (5.94) was not different from that of EFLL (5.95) and tended to be higher than with CON (5.82) or EF (5.82). Postfeeding vitamin B12 concentrations in the rumen were greater with EFSC (134.5ng/g) than with EF (99.6ng/g) and tended to be greater when compared with CON (101.2ng/g) or EFLL (104.9ng/g). During rest, prefeed vitamin B12 was greater with EFSC (166.5ng/g) compared with CON (132.3ng/g). The EFSC treatment did better than EF alone on pH characteristics during adaptation and SARA and on maintenance of ruminal vitamin B12 status during SARA. Milk yield drop from d 1 to 3 of SARA was smaller with EFSC (-0.8kg/d), EF (-0.9kg/d), or EFLL (-0.9kg/d) compared with CON (-7.5kg/d). PMID:25465534

  6. Comparing regions of the Epstein-Barr virus ZEBRA protein which function as transcriptional activating sequences in Saccharomyces cerevisiae and in B cells.

    PubMed Central

    Miller, G; Himmelfarb, H; Heston, L; Countryman, J; Gradoville, L; Baumann, R; Chi, T; Carey, M

    1993-01-01

    The ZEBRA protein activates expression of Epstein-Barr virus early-lytic-cycle genes in human B lymphocytes. Here it is shown that ZEBRA also behaves as a sequence-specific transcriptional activator in Saccharomyces cerevisiae. Deletional mutagenesis defined three regions of ZEBRA that participate in activation in S. cerevisiae. These regions are designated YI (amino acids [aa] 1 to 25), YII (aa 51 to 102), and YIII (aa 228 to 245). Two of the three regions of the native ZEBRA protein act together to mediate activation when assayed on ZEBRA binding sites. However, when fused to the DNA binding domain of GAL4 and assayed on GAL4 binding sites, regions YII and YIII were each sufficient to confer activation in S. cerevisiae. Regions of ZEBRA which affected activation in S. cerevisiae were also required in human B lymphocytes. The amino-terminal region of ZEBRA (aa 1 to 98) was required for activation both in S. cerevisiae and in human B cells; deletion of the carboxy-terminal 18 aa also significantly reduced activation in both cell types. Thus, the behavior of ZEBRA in human B cells and S. cerevisiae suggests that the protein contains universal activation motifs that interact with conserved components of the transcription machinery. However, certain deletion mutants of ZEBRA containing mutations in the N-terminal region exhibited discordant behaviors in S. cerevisiae and in B cells. For example, deletion of ZEBRA aa 26 to 51 impaired activation to a great extent in B cells but had little or no effect in S. cerevisiae. The discordant mutants may reflect interactions with a variable domain of a conserved component or unique interactions with specialized components of the basal transcription apparatus in different cells. PMID:8230468

  7. Fast “Feast/Famine” Cycles for Studying Microbial Physiology Under Dynamic Conditions: A Case Study with Saccharomyces cerevisiae

    PubMed Central

    Suarez-Mendez, Camilo A.; Sousa, Andre; Heijnen, Joseph J.; Wahl, Aljoscha

    2014-01-01

    Microorganisms are constantly exposed to rapidly changing conditions, under natural as well as industrial production scale environments, especially due to large-scale substrate mixing limitations. In this work, we present an experimental approach based on a dynamic feast/famine regime (400 s) that leads to repetitive cycles with moderate changes in substrate availability in an aerobic glucose cultivation of Saccharomyces cerevisiae. After a few cycles, the feast/famine produced a stable and repetitive pattern with a reproducible metabolic response in time, thus providing a robust platform for studying the microorganism’s physiology under dynamic conditions. We found that the biomass yield was slightly reduced (−5%) under the feast/famine regime, while the averaged substrate and oxygen consumption as well as the carbon dioxide production rates were comparable. The dynamic response of the intracellular metabolites showed specific differences in comparison to other dynamic experiments (especially stimulus-response experiments, SRE). Remarkably, the frequently reported ATP paradox observed in single pulse experiments was not present during the repetitive perturbations applied here. We found that intracellular dynamic accumulations led to an uncoupling of the substrate uptake rate (up to 9-fold change at 20 s.) Moreover, the dynamic profiles of the intracellular metabolites obtained with the feast/famine suggest the presence of regulatory mechanisms that resulted in a delayed response. With the feast famine setup many cellular states can be measured at high frequency given the feature of reproducible cycles. The feast/famine regime is thus a versatile platform for systems biology approaches, which can help us to identify and investigate metabolite regulations under realistic conditions (e.g., large-scale bioreactors or natural environments). PMID:24957030

  8. Fast "Feast/Famine" Cycles for Studying Microbial Physiology Under Dynamic Conditions: A Case Study with Saccharomyces cerevisiae.

    PubMed

    Suarez-Mendez, Camilo A; Sousa, Andre; Heijnen, Joseph J; Wahl, Aljoscha

    2014-01-01

    Microorganisms are constantly exposed to rapidly changing conditions, under natural as well as industrial production scale environments, especially due to large-scale substrate mixing limitations. In this work, we present an experimental approach based on a dynamic feast/famine regime (400 s) that leads to repetitive cycles with moderate changes in substrate availability in an aerobic glucose cultivation of Saccharomyces cerevisiae. After a few cycles, the feast/famine produced a stable and repetitive pattern with a reproducible metabolic response in time, thus providing a robust platform for studying the microorganism's physiology under dynamic conditions. We found that the biomass yield was slightly reduced (-5%) under the feast/famine regime, while the averaged substrate and oxygen consumption as well as the carbon dioxide production rates were comparable. The dynamic response of the intracellular metabolites showed specific differences in comparison to other dynamic experiments (especially stimulus-response experiments, SRE). Remarkably, the frequently reported ATP paradox observed in single pulse experiments was not present during the repetitive perturbations applied here. We found that intracellular dynamic accumulations led to an uncoupling of the substrate uptake rate (up to 9-fold change at 20 s.) Moreover, the dynamic profiles of the intracellular metabolites obtained with the feast/famine suggest the presence of regulatory mechanisms that resulted in a delayed response. With the feast famine setup many cellular states can be measured at high frequency given the feature of reproducible cycles. The feast/famine regime is thus a versatile platform for systems biology approaches, which can help us to identify and investigate metabolite regulations under realistic conditions (e.g., large-scale bioreactors or natural environments). PMID:24957030

  9. Single Cell Protein Production by Saccharomyces cerevisiae Using an Optimized Culture Medium Composition in a Batch Submerged Bioprocess.

    PubMed

    Hezarjaribi, Mehrnoosh; Ardestani, Fatemeh; Ghorbani, Hamid Reza

    2016-08-01

    Saccharomyces cerevisiae PTCC5269 growth was evaluated to specify an optimum culture medium to reach the highest protein production. Experiment design was conducted using a fraction of the full factorial methodology, and signal to noise ratio was used for results analysis. Maximum cell of 8.84 log (CFU/mL) was resulted using optimized culture composed of 0.3, 0.15, 1, and 50 g L(-1) of ammonium sulfate, iron sulfate, glycine, and glucose, respectively at 300 rpm and 35 °C. Glycine concentration (39.32 % contribution) and glucose concentration (36.15 % contribution) were determined as the most effective factors on the biomass production, while Saccharomyces cerevisiae growth had showed the least dependence on ammonium sulfate (5.2 % contribution) and iron sulfate (19.28 % contribution). The most interaction was diagnosed between ammonium sulfate and iron sulfate concentrations with interaction severity index of 50.71 %, while the less one recorded for glycine and glucose concentration was equal to 8.12 %. An acceptable consistency of 84.26 % was obtained between optimum theoretical cell numbers determined by software of 8.91 log (CFU/mL), and experimentally measured one at optimal condition confirms the suitability of the applied method. High protein content of 44.6 % using optimum culture suggests that Saccharomyces cerevisiae is a good commercial case for single cell protein production. PMID:27090426

  10. Physiological and transcriptional responses of Saccharomyces cerevisiae to d-limonene show changes to the cell wall but not to the plasma membrane.

    PubMed

    Brennan, Timothy C R; Krömer, Jens O; Nielsen, Lars K

    2013-06-01

    Monoterpenes can, upon hydrogenation, be used as light-fraction components of sustainable aviation fuels. Fermentative production of monoterpenes in engineered microorganisms, such as Saccharomyces cerevisiae, has gained attention as a potential route to deliver these next-generation fuels from renewable biomass. However, end product toxicity presents a formidable problem for microbial synthesis. Due to their hydrophobicity, monoterpene inhibition has long been attributed to membrane interference, but the molecular mechanism remains largely unsolved. In order to gain a better understanding of the mode of action, we analyzed the composition and structural integrity of the cell envelope as well as the transcriptional response of yeast cells treated with an inhibitory amount of d-limonene (107 mg/liter). We found no alterations in membrane fluidity, structural membrane integrity, or fatty acid composition after the solvent challenge. A 4-fold increase in the mean fluorescence intensity per cell (using calcofluor white stain) and increased sensitivity to cell wall-degrading enzymes demonstrated that limonene disrupts cell wall properties. Global transcript measurements confirmed the membrane integrity observations by showing no upregulation of ergosterol or fatty acid biosynthesis pathways, which are commonly overexpressed in yeast to reinforce membrane rigidity during ethanol exposure. Limonene shock did cause a compensatory response to cell wall damage through overexpression of several genes (ROM1, RLM1, PIR3, CTT1, YGP1, MLP1, PST1, and CWP1) involved with the cell wall integrity signaling pathway. This is the first report demonstrating that cell wall, rather than plasma membrane, deterioration is the main source of monoterpene inhibition. We show that limonene can alter the structure and function of the cell wall, which has a clear effect on cytokinesis. PMID:23542628

  11. Physiological and Transcriptional Responses of Saccharomyces cerevisiae to d-Limonene Show Changes to the Cell Wall but Not to the Plasma Membrane

    PubMed Central

    Brennan, Timothy C. R.; Nielsen, Lars K.

    2013-01-01

    Monoterpenes can, upon hydrogenation, be used as light-fraction components of sustainable aviation fuels. Fermentative production of monoterpenes in engineered microorganisms, such as Saccharomyces cerevisiae, has gained attention as a potential route to deliver these next-generation fuels from renewable biomass. However, end product toxicity presents a formidable problem for microbial synthesis. Due to their hydrophobicity, monoterpene inhibition has long been attributed to membrane interference, but the molecular mechanism remains largely unsolved. In order to gain a better understanding of the mode of action, we analyzed the composition and structural integrity of the cell envelope as well as the transcriptional response of yeast cells treated with an inhibitory amount of d-limonene (107 mg/liter). We found no alterations in membrane fluidity, structural membrane integrity, or fatty acid composition after the solvent challenge. A 4-fold increase in the mean fluorescence intensity per cell (using calcofluor white stain) and increased sensitivity to cell wall-degrading enzymes demonstrated that limonene disrupts cell wall properties. Global transcript measurements confirmed the membrane integrity observations by showing no upregulation of ergosterol or fatty acid biosynthesis pathways, which are commonly overexpressed in yeast to reinforce membrane rigidity during ethanol exposure. Limonene shock did cause a compensatory response to cell wall damage through overexpression of several genes (ROM1, RLM1, PIR3, CTT1, YGP1, MLP1, PST1, and CWP1) involved with the cell wall integrity signaling pathway. This is the first report demonstrating that cell wall, rather than plasma membrane, deterioration is the main source of monoterpene inhibition. We show that limonene can alter the structure and function of the cell wall, which has a clear effect on cytokinesis. PMID:23542628

  12. Permeability of the cell envelope and osmotic behavior in Saccharomyces cerevisiae.

    PubMed

    Arnold, W N; Lacy, J S

    1977-08-01

    Bakers' yeast (Saccharomyces cerevisiae) was equilibrated with distilled water and then packed into standardized pellets by centrifugation. The fractional space (S value) that was accessible to passive permeation was probed with a variety of mono- and divalent salts, mono- and disaccharides, polyols, substrates and products of beta-fructofuranosidase (EC 3.2.1.26) and acid phosphatase (EC 3.1.3.2), and a cross-linked polymer of sucrose (Ficoll 400). A simple but very reproducible method was developed to measure pellet volume. At the limit of zero osmolality for bathing medium, the interstitial space was 0.223 ml/ml of pellet, and the aqueous volume of cell envelopes was 0.117 ml/ml of pellet. Thus the cell envelope for this yeast, under these conditions, was approximately 15% of the total cell volume. At a finite osmolality, the space in a yeast pellet that was accessible to salt was accounted for by the sum of initial interstitial space, the volume of the cell envelopes, and the volume of water abstracted from the cells by osmosis. Plots of S value versus osmolality were linear for uncharged probes and curvilinear for all salts. When Ficoll and potassium thiocyanate were presented to the yeast in admixture, the S values for the salt increased continuously over the range of osmolality studied. However, the S values for Ficoll 400 (which did not penetrate the cell wall) were lower by an amount equilivalent to the cell envelopes; they increased in parallel with the S curve for salt up to 1.15 osmol/kg and then plateaued. The results support the concept of incipient plasmolysis at 1.15 osmol/kg, and the separation of protoplasm from the cell wall is indicated with more concentrated solutions. Such cells were still viable if slowly diluted in distilled water, but they were injured by the shock of rapid dilution. However, shocking the cells did not release beta-fructofuranosidase into the medium. The complete accessibility of salts toward killed cells was demonstrated

  13. The Stationary-Phase Cells of Saccharomyces cerevisiae Display Dynamic Actin Filaments Required for Processes Extending Chronological Life Span

    PubMed Central

    Lejskova, Renata; Malcova, Ivana

    2015-01-01

    Stationary-growth-phase Saccharomyces cerevisiae yeast cultures consist of nondividing cells that undergo chronological aging. For their successful survival, the turnover of proteins and organelles, ensured by autophagy and the activation of mitochondria, is performed. Some of these processes are engaged in by the actin cytoskeleton. In S. cerevisiae stationary-phase cells, F actin has been shown to form static aggregates named actin bodies, subsequently cited to be markers of quiescence. Our in vivo analyses revealed that stationary-phase cultures contain cells with dynamic actin filaments, besides the cells with static actin bodies. The cells with dynamic actin displayed active endocytosis and autophagy and well-developed mitochondrial networks. Even more, stationary-phase cell cultures grown under calorie restriction predominantly contained cells with actin cables, confirming that the presence of actin cables is linked to successful adaptation to stationary phase. Cells with actin bodies were inactive in endocytosis and autophagy and displayed aberrations in mitochondrial networks. Notably, cells of the respiratory activity-deficient cox4Δ strain displayed the same mitochondrial aberrations and actin bodies only. Additionally, our results indicate that mitochondrial dysfunction precedes the formation of actin bodies and the appearance of actin bodies corresponds to decreased cell fitness. We conclude that the F-actin status reflects the extent of damage that arises from exponential growth. PMID:26351139

  14. Defects in Protein Folding Machinery Affect Cell Wall Integrity and Reduce Ethanol Tolerance in S. cerevisiae.

    PubMed

    Narayanan, Aswathy; Pullepu, Dileep; Reddy, Praveen Kumar; Uddin, Wasim; Kabir, M Anaul

    2016-07-01

    The chaperonin complex CCT/TRiC (chaperonin containing TCP-1/TCP-1 ring complex) participates in the folding of many crucial proteins including actin and tubulin in eukaryotes. Mutations in genes encoding its subunits can affect protein folding and in turn, the physiology of the organism. Stress response in Saccharomyces cerevisiae is important in fermentation reactions and operates through overexpression and underexpression of genes, thus altering the protein profile. Defective protein folding machinery can disturb this process. In this study, the response of cct mutants to stress conditions in general and ethanol in specific was investigated. CCT1 mutants showed decreased resistance to different conditions tested including osmotic stress, metal ions, surfactants, reducing and oxidising agents. Cct1-3 mutant with the mutation in the conserved ATP-binding region showed irreversible defects than other mutants. These mutants were found to have inherent cell wall defects and showed decreased ethanol tolerance. This study reveals that cell wall defects and ethanol sensitivity are linked. Genetic and proteomic analyses showed that the yeast genes RPS6A (ribosomal protein), SCL1 (proteasomal subunit) and TDH3 (glyceraldehyde-3-phosphate dehydrogenase) on overexpression, improved the growth of cct1-3 mutant on ethanol. We propose the breakdown of common stress response pathways caused by mutations in CCT complex and the resulting scarcity of functional stress-responsive proteins, affecting the cell's defence against different stress agents in cct mutants. Defective cytoskeleton and perturbed cell wall integrity reduce the ethanol tolerance in the mutants which are rescued by the extragenic suppressors. PMID:26992923

  15. Single gene-based distinction of individual microbial genomes from a mixed population of microbial cells

    PubMed Central

    Tamminen, Manu V.; Virta, Marko P. J.

    2015-01-01

    Recent progress in environmental microbiology has revealed vast populations of microbes in any given habitat that cannot be detected by conventional culturing strategies. The use of sensitive genetic detection methods such as CARD-FISH and in situ PCR have been limited by the cell wall permeabilization requirement that cannot be performed similarly on all cell types without lysing some and leaving some nonpermeabilized. Furthermore, the detection of low copy targets such as genes present in single copies in the microbial genomes, has remained problematic. We describe an emulsion-based procedure to trap individual microbial cells into picoliter-volume polyacrylamide droplets that provide a rigid support for genetic material and therefore allow complete degradation of cellular material to expose the individual genomes. The polyacrylamide droplets are subsequently converted into picoliter-scale reactors for genome amplification. The amplified genomes are labeled based on the presence of a target gene and differentiated from those that do not contain the gene by flow cytometry. Using the Escherichia coli strains XL1 and MC1061, which differ with respect to the presence (XL1), or absence (MC1061) of a single copy of a tetracycline resistance gene per genome, we demonstrate that XL1 genomes present at 0.1% of MC1061 genomes can be differentiated using this method. Using a spiked sediment microbial sample, we demonstrate that the method is applicable to highly complex environmental microbial communities as a target gene-based screen for individual microbes. The method provides a novel tool for enumerating functional cell populations in complex microbial communities. We envision that the method could be optimized for fluorescence-activated cell sorting to enrich genetic material of interest from complex environmental samples. PMID:25814987

  16. Evaluation of industrial Saccharomyces cerevisiae strains as the chassis cell for second-generation bioethanol production

    PubMed Central

    Li, Hongxing; Wu, Meiling; Xu, Lili; Hou, Jin; Guo, Ting; Bao, Xiaoming; Shen, Yu

    2015-01-01

    To develop a suitable Saccharomyces cerevisiae industrial strain as a chassis cell for ethanol production using lignocellulosic materials, 32 wild-type strains were evaluated for their glucose fermenting ability, their tolerance to the stresses they might encounter in lignocellulosic hydrolysate fermentation and their genetic background for pentose metabolism. The strain BSIF, isolated from tropical fruit in Thailand, was selected out of the distinctly different strains studied for its promising characteristics. The maximal specific growth rate of BSIF was as high as 0.65 h−1 in yeast extract peptone dextrose medium, and the ethanol yield was 0.45 g g−1 consumed glucose. Furthermore, compared with other strains, this strain exhibited superior tolerance to high temperature, hyperosmotic stress and oxidative stress; better growth performance in lignocellulosic hydrolysate; and better xylose utilization capacity when an initial xylose metabolic pathway was introduced. All of these results indicate that this strain is an excellent chassis strain for lignocellulosic ethanol production. PMID:25616171

  17. Functionally stable and phylogenetically diverse microbial enrichments from microbial fuel cells during wastewater treatment.

    PubMed

    Ishii, Shun'ichi; Suzuki, Shino; Norden-Krichmar, Trina M; Nealson, Kenneth H; Sekiguchi, Yuji; Gorby, Yuri A; Bretschger, Orianna

    2012-01-01

    Microbial fuel cells (MFCs) are devices that exploit microorganisms as biocatalysts to recover energy from organic matter in the form of electricity. One of the goals of MFC research is to develop the technology for cost-effective wastewater treatment. However, before practical MFC applications are implemented it is important to gain fundamental knowledge about long-term system performance, reproducibility, and the formation and maintenance of functionally-stable microbial communities. Here we report findings from a MFC operated for over 300 days using only primary clarifier effluent collected from a municipal wastewater treatment plant as the microbial resource and substrate. The system was operated in a repeat-batch mode, where the reactor solution was replaced once every two weeks with new primary effluent that consisted of different microbial and chemical compositions with every batch exchange. The turbidity of the primary clarifier effluent solution notably decreased, and 97% of biological oxygen demand (BOD) was removed after an 8-13 day residence time for each batch cycle. On average, the limiting current density was 1000 mA/m(2), the maximum power density was 13 mW/m(2), and coulombic efficiency was 25%. Interestingly, the electrochemical performance and BOD removal rates were very reproducible throughout MFC operation regardless of the sample variability associated with each wastewater exchange. While MFC performance was very reproducible, the phylogenetic analyses of anode-associated electricity-generating biofilms showed that the microbial populations temporally fluctuated and maintained a high biodiversity throughout the year-long experiment. These results suggest that MFC communities are both self-selecting and self-optimizing, thereby able to develop and maintain functional stability regardless of fluctuations in carbon source(s) and regular introduction of microbial competitors. These results contribute significantly toward the practical application

  18. Functionally Stable and Phylogenetically Diverse Microbial Enrichments from Microbial Fuel Cells during Wastewater Treatment

    PubMed Central

    Ishii, Shun'ichi; Suzuki, Shino; Norden-Krichmar, Trina M.; Nealson, Kenneth H.; Sekiguchi, Yuji; Gorby, Yuri A.; Bretschger, Orianna

    2012-01-01

    Microbial fuel cells (MFCs) are devices that exploit microorganisms as biocatalysts to recover energy from organic matter in the form of electricity. One of the goals of MFC research is to develop the technology for cost-effective wastewater treatment. However, before practical MFC applications are implemented it is important to gain fundamental knowledge about long-term system performance, reproducibility, and the formation and maintenance of functionally-stable microbial communities. Here we report findings from a MFC operated for over 300 days using only primary clarifier effluent collected from a municipal wastewater treatment plant as the microbial resource and substrate. The system was operated in a repeat-batch mode, where the reactor solution was replaced once every two weeks with new primary effluent that consisted of different microbial and chemical compositions with every batch exchange. The turbidity of the primary clarifier effluent solution notably decreased, and 97% of biological oxygen demand (BOD) was removed after an 8–13 day residence time for each batch cycle. On average, the limiting current density was 1000 mA/m2, the maximum power density was 13 mW/m2, and coulombic efficiency was 25%. Interestingly, the electrochemical performance and BOD removal rates were very reproducible throughout MFC operation regardless of the sample variability associated with each wastewater exchange. While MFC performance was very reproducible, the phylogenetic analyses of anode-associated electricity-generating biofilms showed that the microbial populations temporally fluctuated and maintained a high biodiversity throughout the year-long experiment. These results suggest that MFC communities are both self-selecting and self-optimizing, thereby able to develop and maintain functional stability regardless of fluctuations in carbon source(s) and regular introduction of microbial competitors. These results contribute significantly toward the practical application of

  19. Simultaneous microbial and electrochemical reductions of vanadium (V) with bioelectricity generation in microbial fuel cells.

    PubMed

    Zhang, Baogang; Tian, Caixing; Liu, Ying; Hao, Liting; Liu, Ye; Feng, Chuanping; Liu, Yuqian; Wang, Zhongli

    2015-03-01

    Simultaneous microbial and electrochemical reductions of vanadium (V) with bioelectricity generation were realized in microbial fuel cells (MFCs). With initial V(V) concentrations of 75 mg/l and 150 mg/l in anolyte and catholyte, respectively, stable power output of 419±11 mW/m(2) was achieved. After 12h operation, V(V) concentration in the catholyte decreased to the value similar to that of the initial one in the anolyte, meanwhile it was nearly reduced completely in the anolyte. V(IV) was the main reduction product, which subsequently precipitated, acquiring total vanadium removal efficiencies of 76.8±2.9%. Microbial community analysis revealed the emergence of the new species of Deltaproteobacteria and Bacteroidetes as well as the enhanced Spirochaetes mainly functioned in the anode. This study opens new pathways to successful remediation of vanadium contamination. PMID:25536507

  20. Effects of light intensity and titanium dioxide concentration on photocatalytic sterilization rates of microbial cells

    SciTech Connect

    Horie, Yasuhiko; David, D.A.1; Taya, Masahito; Tone, Setsuji

    1996-11-01

    Photocatalytic sterilization of Escherichia coli (bacterium) or Saccharomyces serevisiae (yeast) was conducted with a rectangular bubble-column photoreactor (40 mm in width, 40 mm in breadth, and 250 mm in height) containing slurried TiO{sub 2} semiconductor particles. The profiles of cell deactivation with sterilization time could be expressed in fair agreement with experimental data, based on a series-event model and a second-order kinetics with respect to the concentrations of microbial cells and oxidative radicals generated by photoexcitation of TiO{sub 2} particles. Sterilization rate constants for the microbes were determined under various conditions of TiO{sub 2} concentrations (0--5 {times} 0{sup {minus}1} kg/m{sup 3}) and average light intensities (0--223 W/m{sup 2}) in the photoreactor. Linear relationships were obtained between the rate constants and average light intensity at TiO{sub 2} concentration of 1 {times} 10{sup {minus}2} kg/m{sup 3}. When incident light intensity was kept constant (27 W/m{sup 2} for E. coli or 238 W/m{sup 2} for S. cerevisiae), the correlations between the rate constants and TiO{sub 2} concentration were interpreted considering a fraction of TiO{sub 2} particles adhered to the cells in slurry.

  1. Evolutionary engineering of Saccharomyces cerevisiae for efficient aerobic xylose consumption.

    PubMed

    Scalcinati, Gionata; Otero, José Manuel; Van Vleet, Jennifer R H; Jeffries, Thomas W; Olsson, Lisbeth; Nielsen, Jens

    2012-08-01

    Industrial biotechnology aims to develop robust microbial cell factories, such as Saccharomyces cerevisiae, to produce an array of added value chemicals presently dominated by petrochemical processes. Xylose is the second most abundant monosaccharide after glucose and the most prevalent pentose sugar found in lignocelluloses. Significant research efforts have focused on the metabolic engineering of S. cerevisiae for fast and efficient xylose utilization. This study aims to metabolically engineer S. cerevisiae, such that it can consume xylose as the exclusive substrate while maximizing carbon flux to biomass production. Such a platform may then be enhanced with complementary metabolic engineering strategies that couple biomass production with high value-added chemical. Saccharomyces cerevisiae, expressing xylose reductase, xylitol dehydrogenase and xylulose kinase, from the native xylose-metabolizing yeast Pichia stipitis, was constructed, followed by a directed evolution strategy to improve xylose utilization rates. The resulting S. cerevisiae strain was capable of rapid growth and fast xylose consumption producing only biomass and negligible amount of byproducts. Transcriptional profiling of this strain was employed to further elucidate the observed physiology confirms a strongly up-regulated glyoxylate pathway enabling respiratory metabolism. The resulting strain is a desirable platform for the industrial production of biomass-related products using xylose as a sole carbon source. PMID:22487265

  2. Recognition of Microbial Glycolipids by Natural Killer T Cells

    PubMed Central

    Zajonc, Dirk M.; Girardi, Enrico

    2015-01-01

    T cells can recognize microbial antigens when presented by dedicated antigen-presenting molecules. While peptides are presented by classical members of the major histocompatibility complex (MHC) family (MHC I and II), lipids, glycolipids, and lipopeptides can be presented by the non-classical MHC member, CD1. The best studied subset of lipid-reactive T cells are type I natural killer T (iNKT) cells that recognize a variety of different antigens when presented by the non-classical MHCI homolog CD1d. iNKT cells have been shown to be important for the protection against various microbial pathogens, including B. burgdorferi, the causative agents of Lyme disease, and S. pneumoniae, which causes pneumococcal meningitis and community-acquired pneumonia. Both pathogens carry microbial glycolipids that can trigger the T cell antigen receptor (TCR), leading to iNKT cell activation. iNKT cells have an evolutionary conserved TCR alpha chain, yet retain the ability to recognize structurally diverse glycolipids. They do so using a conserved recognition mode, in which the TCR enforces a conserved binding orientation on CD1d. TCR binding is accompanied by structural changes within the TCR binding site of CD1d, as well as the glycolipid antigen itself. In addition to direct recognition of microbial antigens, iNKT cells can also be activated by a combination of cytokines (IL-12/IL-18) and TCR stimulation. Many microbes carry TLR antigens, and microbial infections can lead to TLR activation. The subsequent cytokine response in turn lower the threshold of TCR-mediated iNKT cell activation, especially when weak microbial or even self-antigens are presented during the cause of the infection. In summary, iNKT cells can be directly activated through TCR triggering of strong antigens, while cytokines produced by the innate immune response may be necessary for TCR triggering and iNKT cell activation in the presence of weak antigens. Here, we will review the molecular basis of iNKT cell

  3. The Transient Inactivation of the Master Cell Cycle Phosphatase Cdc14 Causes Genomic Instability in Diploid Cells of Saccharomyces cerevisiae.

    PubMed

    Quevedo, Oliver; Ramos-Pérez, Cristina; Petes, Thomas D; Machín, Félix

    2015-07-01

    Genomic instability is a common feature found in cancer cells . Accordingly, many tumor suppressor genes identified in familiar cancer syndromes are involved in the maintenance of the stability of the genome during every cell division and are commonly referred to as caretakers. Inactivating mutations and epigenetic silencing of caretakers are thought to be the most important mechanisms that explain cancer-related genome instability. However, little is known of whether transient inactivation of caretaker proteins could trigger genome instability and, if so, what types of instability would occur. In this work, we show that a brief and reversible inactivation, during just one cell cycle, of the key phosphatase Cdc14 in the model organism Saccharomyces cerevisiae is enough to result in diploid cells with multiple gross chromosomal rearrangements and changes in ploidy. Interestingly, we observed that such transient loss yields a characteristic fingerprint whereby trisomies are often found in small-sized chromosomes, and gross chromosome rearrangements, often associated with concomitant loss of heterozygosity, are detected mainly on the ribosomal DNA-bearing chromosome XII. Taking into account the key role of Cdc14 in preventing anaphase bridges, resetting replication origins, and controlling spindle dynamics in a well-defined window within anaphase, we speculate that the transient loss of Cdc14 activity causes cells to go through a single mitotic catastrophe with irreversible consequences for the genome stability of the progeny. PMID:25971663

  4. The Transient Inactivation of the Master Cell Cycle Phosphatase Cdc14 Causes Genomic Instability in Diploid Cells of Saccharomyces cerevisiae

    PubMed Central

    Quevedo, Oliver; Ramos-Pérez, Cristina; Petes, Thomas D.; Machín, Félix

    2015-01-01

    Genomic instability is a common feature found in cancer cells . Accordingly, many tumor suppressor genes identified in familiar cancer syndromes are involved in the maintenance of the stability of the genome during every cell division and are commonly referred to as caretakers. Inactivating mutations and epigenetic silencing of caretakers are thought to be the most important mechanisms that explain cancer-related genome instability. However, little is known of whether transient inactivation of caretaker proteins could trigger genome instability and, if so, what types of instability would occur. In this work, we show that a brief and reversible inactivation, during just one cell cycle, of the key phosphatase Cdc14 in the model organism Saccharomyces cerevisiae is enough to result in diploid cells with multiple gross chromosomal rearrangements and changes in ploidy. Interestingly, we observed that such transient loss yields a characteristic fingerprint whereby trisomies are often found in small-sized chromosomes, and gross chromosome rearrangements, often associated with concomitant loss of heterozygosity, are detected mainly on the ribosomal DNA-bearing chromosome XII. Taking into account the key role of Cdc14 in preventing anaphase bridges, resetting replication origins, and controlling spindle dynamics in a well-defined window within anaphase, we speculate that the transient loss of Cdc14 activity causes cells to go through a single mitotic catastrophe with irreversible consequences for the genome stability of the progeny. PMID:25971663

  5. Significant quantities of the glycolytic enzyme phosphoglycerate mutase are present in the cell wall of yeast Saccharomyces cerevisiae.

    PubMed Central

    Motshwene, Precious; Brandt, Wolf; Lindsey, George

    2003-01-01

    NaOH was used to extract proteins from the cell walls of the yeast Saccharomyces cerevisiae. This treatment was shown not to disrupt yeast cells, as NaOH-extracted cells displayed a normal morphology upon electron microscopy. Moreover, extracted and untreated cells had qualitatively similar protein contents upon disruption. When yeast was grown in the presence of 1 M mannitol, two proteins were found to be present at an elevated concentration in the cell wall. These were found to be the late-embryogenic-abundant-like protein heat-shock protein 12 and the glycolytic enzyme phosphoglycerate mutase. The presence of phosphoglycerate mutase in the cell wall was confirmed by immunocytochemical analysis. Not only was the phosphoglycerate mutase in the yeast cell wall found to be active, but whole yeast cells were also able to convert 3-phosphoglycerate in the medium into ethanol, provided that the necessary cofactors were present. PMID:12238949

  6. CD1-Restricted T Cell Recognition of Microbial Lipoglycan Antigens

    NASA Astrophysics Data System (ADS)

    Sieling, P. A.; Chatterjee, D.; Porcelli, S. A.; Prigozy, T. I.; Mazzaccaro, R. J.; Soriano, T.; Bloom, B. R.; Brenner, M. B.; Kronenberg, M.; Brennan, P. J.; Modlin, R. L.

    1995-07-01

    It has long been the paradigm that T cells recognize peptide antigens presented by major histocompatibility complex (MHC) molecules. However, nonpeptide antigens can be presented to T cells by human CD1b molecules, which are not encoded by the MHC. A major class of microbial antigens associated with pathogenicity are lipoglycans. It is shown here that human CD1b presents the defined mycobacterial lipoglycan lipoarabinomannan (LAM) to αβ T cell receptor-bearing lymphocytes. Presentation of these lipoglycan antigens required internalization and endosomal acidification. The T cell recognition required mannosides with α(1-->2) linkages and a phosphatidylinositol unit. T cells activated by LAM produced interferon γ and were cytolytic. Thus, an important class of microbial molecules, the lipoglycans, is a part of the universe of foreign antigens recognized by human T cells.

  7. Impact of cell density on microbially induced stable isotope fractionation.

    PubMed

    Kampara, Makeba; Thullner, Martin; Harms, Hauke; Wick, Lukas Y

    2009-01-01

    Quantification of microbial contaminant biodegradation based on stable isotope fractionation analysis (SIFA) relies on known, invariable isotope fractionation factors. The microbially induced isotope fractionation is caused by the preferential cleavage of bonds containing light rather than heavy isotopes. However, a number of non-isotopically sensitive steps preceding the isotopically sensitive bond cleavage may affect the reaction kinetics of a degradation process and reduce the observed (i.e., the macroscopically detectable) isotope fractionation. This introduces uncertainty to the use of isotope fractionation for the quantification of microbial degradation processes. Here, we report on the influence of bacterial cell density on observed stable isotope fractionation. Batch biodegradation experiments were performed under non-growth conditions to quantify the toluene hydrogen isotope fractionation by exposing Pseudomonas putida mt-2(pWWO) at varying cell densities to different concentrations of toluene. Observed isotope fractionation depended significantly on the cell density. When the cell density rose from 5 x 10(5) to 5 x 10(8)cells/mL, the observed isotope fractionation declined by 70% and went along with a 55% decrease of the degradation rates of individual cells. Theoretical estimates showed that uptake-driven diffusion to individual cells depended on cell density via the overlap of the cells' diffusion-controlled boundary layers. Our data suggest that biomass effects on SIFA have to be considered even in well-mixed systems such as the cell suspensions used in this study. PMID:19015849

  8. Bioethanol production from mixed sugars by Scheffersomyces stipitis free and immobilized cells, and co-cultures with Saccharomyces cerevisiae.

    PubMed

    De Bari, Isabella; De Canio, Paola; Cuna, Daniela; Liuzzi, Federico; Capece, Angela; Romano, Patrizia

    2013-09-25

    Bioethanol can be produced from several biomasses including lignocellulosic materials. Besides 6-carbon sugars that represent the prevalent carbohydrates, some of these feedstocks contain significant amounts of 5-carbon sugars. One common limit of the major part of the xylose-fermenting yeasts is the diauxic shift between the uptake of glucose and xylose during the fermentation of mixed syrups. Thus, optimized fermentation strategies are required. In this paper the ability of Scheffersomyces stipitis strain NRRLY-11544 to ferment mixed syrups with a total sugar concentration in the range 40-80 g/L was investigated by using mono cultures, co-cultures with Saccharomyces cerevisiae strain Bakers Yeast Type II and single cultures immobilized in silica-hydrogel films. The experimental design for the fermentations with immobilized cells included the process analysis in function of two parameters: the fraction of the gel in the broth and the concentration of the cells loaded in the gel. Furthermore, for each total sugars level, the fermentative course of S. stipitis was analyzed at several glucose-to xylose ratios. The results indicated that the use of S. stipitis and S. cerevisiae in free co-cultures ensured faster processes than single cultures of S. stipitis either free or immobilized. However, the rapid production of ethanol by S. cerevisiae inhibited S. stipitis and caused a stuck of the process. Immobilization of S. stipitis in silica-hydrogel increased the relative consumption rate of xylose-to-glucose by 2-6 times depending on the composition of the fermentation medium. Furthermore the films performances appeared stable over three weeks of continuous operations. However, on the whole, the final process yields obtained with the immobilized cells were not meaningfully different from that of the free cells. This was probably due to concurrent fermentations operated by the cells released in the broth. Optimization of the carrier characteristics could improve the

  9. Identification of autophagy genes participating in zinc-induced necrotic cell death in Saccharomyces cerevisiae

    PubMed Central

    Dziedzic, Slawomir A

    2011-01-01

    Eukaryotes use a common set of genes to perform two mechanistically similar autophagic processes. Bulk autophagy harvests proteins nonselectively and reuses their constitutents when nutrients are scarce. In contrast, different forms of selective autophagy target protein aggregates or damaged organelles that threaten to interfere with growth. Yeast uses one form of selective autophagy, called cytoplasm-to-vacuole targeting (Cvt), to engulf two vacuolar enzymes in Cvt vesicles (“CVT-somes”) within which they are transported to vacuoles for maturation. While both are dispensable normally, bulk and selective autophagy help sustain life under stressful conditions. Consistent with this view, knocking out several genes participating in Cvt and specialized autophagic pathways heightened the sensitivity of Saccharomyces cerevisiae to inhibitory levels of Zn2+. The loss of other autophagic genes, and genes responsible for apoptotic cell death, had no such effect. Unexpectedly, the loss of members of a third set of autophagy genes heightened cellular resistance to zinc as if they encoded proteins that actively contributed to zinc-induced cell death. Further studies showed that both sensitive and resistant strains accumulated similar amounts of H2O2 during zinc treatments, but that more sensitive strains showed signs of necrosis sooner. Although zinc lethality depended on autophagic proteins, studies with several reporter genes failed to reveal increased autophagic activity. In fact, microscopy analysis indicated that Zn2+ partially inhibited fusion of Cvt vesicles with vacuoles. Further studies into how the loss of autophagic processes suppressed necrosis in yeast might reveal whether a similar process could occur in plants and animals. PMID:21317551

  10. Recent advances in microbial single cell genomics technology and applications

    NASA Astrophysics Data System (ADS)

    Stepanauskas, R.

    2015-12-01

    Single cell genomics is increasingly utilized as a powerful tool to decipher the metabolic potential, evolutionary histories and in situ interactions of environmental microorganisms. I will present several new developments of this exciting technology, which improve genomic data recovery from individual cells and allow its integration with cell's phenotypic properties. I will also demonstrate how these new technical capabilities help understanding the biology of the "microbial dark matter" inhabiting marine and terrestrial subsurface environments.

  11. Cell Surface Display of Four Types of Solanum nigrum Metallothionein on Saccharomyces cerevisiae for Biosorption of Cadmium.

    PubMed

    Wei, Qinguo; Zhang, Honghai; Guo, Dongge; Ma, Shisheng

    2016-05-28

    We displayed four types of Solanum nigrum metallothionein (SMT) for the first time on the surface of Saccharomyces cerevisiae using an α-agglutinin-based display system. The SMT genes were amplified by RT-PCR. The plasmid pYES2 was used to construct the expression vector. Transformed yeast strains were confirmed by PCR amplification and custom sequencing. Surface-expressed metallothioneins were indirectly indicated by the enhanced cadmium sorption capacity. Flame atomic absorption spectrophotometry was used to examine the concentration of Cd(2+) in this study. The transformed yeast strains showed much higher resistance ability to Cd(2+) compared with the control. Strikingly, their Cd(2+) accumulation was almost twice as much as that of the wild-type yeast cells. Furthermore, surface-engineered yeast strains could effectively adsorb ultra-trace cadmium and accumulate Cd(2+) under a wide range of pH levels, from 3 to 7, without disturbing the Cu(2+) and Hg(2+). Four types of surfaceengineered Saccharomyces cerevisiae strains were constructed and they could be used to purify Cd(2+)-contaminated water and adsorb ultra-trace cadmium effectively. The surface-engineered Saccharomyces cerevisiae strains would be useful tools for the bioremediation and biosorption of environmental cadmium contaminants. PMID:26838339

  12. Rapid Identification and Enumeration of Saccharomyces cerevisiae Cells in Wine by Real-Time PCR

    PubMed Central

    Martorell, P.; Querol, A.; Fernández-Espinar, M. T.

    2005-01-01

    Despite the beneficial role of Saccharomyces cerevisiae in the food industry for food and beverage production, it is able to cause spoilage in wines. We have developed a real-time PCR method to directly detect and quantify this yeast species in wine samples to provide winemakers with a rapid and sensitive method to detect and prevent wine spoilage. Specific primers were designed for S. cerevisiae using the sequence information obtained from a cloned random amplified polymorphic DNA band that differentiated S. cerevisiae from its sibling species Saccharomyces bayanus, Saccharomyces pastorianus, and Saccharomyces paradoxus. The specificity of the primers was demonstrated for typical wine spoilage yeast species. The method was useful for estimating the level of S. cerevisiae directly in sweet wines and red wines without preenrichment when yeast is present in concentrations as low as 3.8 and 5 CFU per ml. This detection limit is in the same order as that obtained from glucose-peptone-yeast growth medium (GPY). Moreover, it was possible to quantify S. cerevisiae in artificially contaminated samples accurately. Limits for accurate quantification in wine were established, from 3.8 × 105 to 3.8 CFU/ml in sweet wine and from 5 × 106 to 50 CFU/ml in red wine. PMID:16269715

  13. Segregation of the Anodic Microbial Communities in a Microbial Fuel Cell Cascade

    PubMed Central

    Hodgson, Douglas M.; Smith, Ann; Dahale, Sonal; Stratford, James P.; Li, Jia V.; Grüning, André; Bushell, Michael E.; Marchesi, Julian R.; Avignone Rossa, C.

    2016-01-01

    Metabolic interactions within microbial communities are essential for the efficient degradation of complex organic compounds, and underpin natural phenomena driven by microorganisms, such as the recycling of carbon-, nitrogen-, and sulfur-containing molecules. These metabolic interactions ultimately determine the function, activity and stability of the community, and therefore their understanding would be essential to steer processes where microbial communities are involved. This is exploited in the design of microbial fuel cells (MFCs), bioelectrochemical devices that convert the chemical energy present in substrates into electrical energy through the metabolic activity of microorganisms, either single species or communities. In this work, we analyzed the evolution of the microbial community structure in a cascade of MFCs inoculated with an anaerobic microbial community and continuously fed with a complex medium. The analysis of the composition of the anodic communities revealed the establishment of different communities in the anodes of the hydraulically connected MFCs, with a decrease in the abundance of fermentative taxa and a concurrent increase in respiratory taxa along the cascade. The analysis of the metabolites in the anodic suspension showed a metabolic shift between the first and last MFC, confirming the segregation of the anodic communities. Those results suggest a metabolic interaction mechanism between the predominant fermentative bacteria at the first stages of the cascade and the anaerobic respiratory electrogenic population in the latter stages, which is reflected in the observed increase in power output. We show that our experimental system represents an ideal platform for optimization of processes where the degradation of complex substrates is involved, as well as a potential tool for the study of metabolic interactions in complex microbial communities. PMID:27242723

  14. Segregation of the Anodic Microbial Communities in a Microbial Fuel Cell Cascade.

    PubMed

    Hodgson, Douglas M; Smith, Ann; Dahale, Sonal; Stratford, James P; Li, Jia V; Grüning, André; Bushell, Michael E; Marchesi, Julian R; Avignone Rossa, C

    2016-01-01

    Metabolic interactions within microbial communities are essential for the efficient degradation of complex organic compounds, and underpin natural phenomena driven by microorganisms, such as the recycling of carbon-, nitrogen-, and sulfur-containing molecules. These metabolic interactions ultimately determine the function, activity and stability of the community, and therefore their understanding would be essential to steer processes where microbial communities are involved. This is exploited in the design of microbial fuel cells (MFCs), bioelectrochemical devices that convert the chemical energy present in substrates into electrical energy through the metabolic activity of microorganisms, either single species or communities. In this work, we analyzed the evolution of the microbial community structure in a cascade of MFCs inoculated with an anaerobic microbial community and continuously fed with a complex medium. The analysis of the composition of the anodic communities revealed the establishment of different communities in the anodes of the hydraulically connected MFCs, with a decrease in the abundance of fermentative taxa and a concurrent increase in respiratory taxa along the cascade. The analysis of the metabolites in the anodic suspension showed a metabolic shift between the first and last MFC, confirming the segregation of the anodic communities. Those results suggest a metabolic interaction mechanism between the predominant fermentative bacteria at the first stages of the cascade and the anaerobic respiratory electrogenic population in the latter stages, which is reflected in the observed increase in power output. We show that our experimental system represents an ideal platform for optimization of processes where the degradation of complex substrates is involved, as well as a potential tool for the study of metabolic interactions in complex microbial communities. PMID:27242723

  15. Cyclin-Dependent Kinase Co-Ordinates Carbohydrate Metabolism and Cell Cycle in S. cerevisiae.

    PubMed

    Zhao, Gang; Chen, Yuping; Carey, Lucas; Futcher, Bruce

    2016-05-19

    Cyclin-dependent kinases (CDKs) control cell division in eukaryotes by phosphorylating proteins involved in division. But successful proliferation requires co-ordination between division and cellular growth in mass. Previous proteomic studies suggested that metabolic proteins, as well as cell division proteins, could potentially be substrates of cyclin-dependent kinases. Here we focus on two metabolic enzymes of the yeast S. cerevisiae, neutral trehalase (Nth1) and glycogen phosphorylase (Gph1), and show that their activities are likely directly controlled by CDK activity, thus allowing co-ordinate regulation of carbohydrate metabolism with cell division processes. In this case, co-ordinate regulation may optimize the decision to undertake a final cell division as nutrients are being exhausted. Co-regulation of cell division processes and metabolic processes by CDK activity may be a general phenomenon important for co-ordinating the cell cycle with growth. PMID:27203179

  16. Comparison of cell wall proteins of Saccharomyces cerevisiae as anchors for cell surface expression of heterologous proteins.

    PubMed Central

    Van der Vaart, J M; te Biesebeke, R; Chapman, J W; Toschka, H Y; Klis, F M; Verrips, C T

    1997-01-01

    The carboxyl-terminal regions of five cell wall proteins (Cwp1p, Cwp2p, Ag alpha 1p, Tip1p, and Flo1p) and three potential cell wall proteins (Sed1p, YCR89w, and Tir1p) all proved capable of immobilizing alpha-galactosidase in the cell wall of Saccharomyces cerevisiae. The fraction of the total amount of fusion protein that was localized to the cell wall varied depending on the anchor domain used. The highest proportion of cell wall incorporation was achieved with Cwp2p, Ag alpha 1p, or Sed1p as an anchor. Although 80% of these fusion proteins were incorporated in the cell wall, the total production of alpha-galactosidase-Ag alpha 1p was sixfold lower than that of alpha-galactosidase-Cwp2p and eightfold lower than that of alpha-galactosidase-Sed1p. Differences in mRNA levels were not responsible for this discrepancy, nor was an intracellular accumulation of alpha-galactosidase-Ag alpha 1p detectable. A lower translation efficiency of the alpha-galactosidase-AG alpha 1 fusion construct is most likely to be responsible for the low level of protein production. alpha-Galactosidase immobilized by the carboxyl-terminal 67 amino acids of Cwp2p was most effective in the hydrolysis of the high-molecular-weight substrate guar gum from Cyamopsis tetragonoloba. This indicates that the use of a large anchoring domain does not necessarily result in a better exposure of the immobilized enzyme to the exterior of the yeast cell. PMID:9023939

  17. The Microbial Fuel Cell as an Education Tool

    ERIC Educational Resources Information Center

    Dewan, Alim; Van Wie, Bernard; Beyenal, Haluk; Lewandowski, Zbigniew

    2010-01-01

    Many chemical engineering programs offer courses from a variety of disciplines to teach their students multidisciplinary concepts, but often these courses lack appropriate tools for linking newly learned concepts to principles learned in the core courses. This paper describes our experience of incorporating a microbial fuel cell education module…

  18. Microbial Fuel Cell Performance with a Pressurized Cathode Chamber

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Microbial fuel cell (MFC) power densities are often constrained by the oxygen reduction reaction rate on the cathode electrode. One important factor for this is the normally low solubility of oxygen in the aqueous cathode solution creating mass transport limitations, which hinder oxygen reduction a...

  19. Oxygen - Enemy or Friend for Microbial Fuel Cell Anode Performance?

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Until recently, scientists and engineers have held a strong belief that oxygen intrusion into the anode chamber of a bioelectrochemical system (BES) is detrimental to microbial fuel cell (MFC) performance because oxygen acts as an alternate electron acceptor. This would, according to recent beliefs...

  20. The steady-state level and stability of TLS polymerase eta are cell cycle dependent in the yeast S. cerevisiae.

    PubMed

    Plachta, Michal; Halas, Agnieszka; McIntyre, Justyna; Sledziewska-Gojska, Ewa

    2015-05-01

    Polymerase eta (Pol eta) is a ubiquitous translesion DNA polymerase that is capable of bypassing UV-induced pyrimidine dimers in an error-free manner. However, this specialized polymerase is error prone when synthesizing through an undamaged DNA template. In Saccharomyces cerevisiae, both depletion and overproduction of Pol eta result in mutator phenotypes. Therefore, regulation of the cellular abundance of this enzyme is of particular interest. However, based on the investigation of variously tagged forms of Pol eta, mutually contradictory conclusions have been reached regarding the stability of this polymerase in yeast. Here, we optimized a protocol for the detection of untagged yeast Pol eta and established that the half-life of the native enzyme is 80 ± 14 min in asynchronously growing cultures. Experiments with synchronized cells indicated that the cellular abundance of this translesion polymerase changes throughout the cell cycle. Accordingly, we show that the stability of Pol eta, but not its mRNA level, is cell cycle stage dependent. The half-life of the polymerase is more than fourfold shorter in G1-arrested cells than in those at G2/M. Our results, in concert with previous data for Rev1, indicate that cell cycle regulation is a general property of Y family TLS polymerases in S. cerevisiae. PMID:25766643

  1. Microbial community dynamics in continuous microbial fuel cells fed with synthetic wastewater and pig slurry.

    PubMed

    Sotres, Ana; Tey, Laura; Bonmatí, August; Viñas, Marc

    2016-10-01

    Two-chambered microbial fuel cells (MFCs) operating with synthetic wastewater and pig slurry were assessed. Additionally, the use of 2-bromoethanesulfonate (BES-Inh) was studied. The synthetic wastewater-fed MFC (MFCSW) showed a maximum power density (PDmax) of 2138mWm(-3), and the addition of BES-Inh (10mM) did not show any improvement in its performance (PDmax=2078mWm(-3)). When pig slurry was used as feed (MFCPS), PDmax increased up to 5623mWm(-3). The microbial community composition was affected by the type of substrate used. While, Pseudomonadaceae and Clostridiaceae were the most representative families within the acetate-based medium, Flavobacteriaceae, Chitinophagaceae, Comamonadaceae and Nitrosomonadaceae were predominant when pig slurry was used as feed. Otherwise, only the Eubacterial microbial community composition was strongly modified when adding BES-Inh, thus leading to an enrichment of the Bacteroidetes phylum. Oppositely, the Archaeal community was less affected by the addition of BES-Inh, and Methanosarcina sp., arose as the predominant family in both situations. Despite all the differences in microbial communities, 6 operational taxonomic units (OTUs) belonging to Bacteroidetes (Porphyromonadaceae and Marinilabiaceae) and Firmicutes (Clostridiales) were found to be common to both MFCs, also for different contents of COD and N-NH4(+), and therefore could be considered as the bioanode core microbiome. PMID:27243446

  2. Multi-omics data driven analysis establishes reference codon biases for synthetic gene design in microbial and mammalian cells.

    PubMed

    Ang, Kok Siong; Kyriakopoulos, Sarantos; Li, Wei; Lee, Dong-Yup

    2016-06-01

    In this study, we analyzed multi-omics data and subsets thereof to establish reference codon usage biases for codon optimization in synthetic gene design. Specifically, publicly available genomic, transcriptomic, proteomic and translatomic data for microbial and mammalian expression hosts, Escherichia coli, Saccharomyces cerevisiae, Pichia pastoris and Chinese hamster ovary (CHO) cells, were compiled to derive their individual codon and codon pair frequencies. Then, host dependent and -omics specific codon biases were generated and compared by principal component analysis and hierarchical clustering. Interestingly, our results indicated the similar codon bias patterns of the highly expressed transcripts, highly abundant proteins, and efficiently translated mRNA in microbial cells, despite the general lack of correlation between mRNA and protein expression levels. However, for CHO cells, the codon bias patterns among various -omics subsets are not distinguishable, forming one cluster. Thus, we further investigated the effect of different input codon biases on codon optimized sequences using the codon context (CC) and individual codon usage (ICU) design parameters, via in silico case study on the expression of human IFNγ sequence in CHO cells. The results supported that CC is more robust design parameter than ICU for improved heterologous gene design. PMID:26850284

  3. Microbial fuel cell treatment of fuel process wastewater

    DOEpatents

    Borole, Abhijeet P; Tsouris, Constantino

    2013-12-03

    The present invention is directed to a method for cleansing fuel processing effluent containing carbonaceous compounds and inorganic salts, the method comprising contacting the fuel processing effluent with an anode of a microbial fuel ell, the anode containing microbes thereon which oxidatively degrade one or more of the carbonaceous compounds while producing electrical energy from the oxidative degradation, and directing the produced electrical energy to drive an electrosorption mechanism that operates to reduce the concentration of one or more inorganic salts in the fuel processing effluent, wherein the anode is in electrical communication with a cathode of the microbial fuel cell. The invention is also directed to an apparatus for practicing the method.

  4. Coastal microbial fuel cell: scaling laws and systems

    NASA Astrophysics Data System (ADS)

    Bandyopadhyay, Promode R.; McNeilly, Frank J.; Thivierge, Daniel P.; Fredette, Albert R.

    2006-05-01

    Microbes, like Geobacters, have inhabited the seafloors around the world since the early days of earth. Such regions are anaerobic and they gain energy by using the widely prevalent iron oxides and organic matters. Because they appear to colonize conducting surfaces that act as sinks of electrons, microbial fuel cells have been shown to convert organic matter to electricity. A microbial fuel cell system has been deployed in Narragansett Bay in Newport, Rhode Island for a year. Currently, the cathode and anode areas are of the order of that of a small wind mill. Measurements have been carried out to determine the marine scaling laws of power harvesting in passive benthic microbial fuel cells. The focus has been on the ocean engineering aspects such as marine scaling laws and the integration of the biochemical and the electronic systems. The characteristics examined are: the relationship of electrode surface area and power produced, the stabilization rates of ionic paths, that is, the effects of location depth of cathodes on stabilization after deployment, the effects of solar and lunar cycles in the Narragansett Bay on the dynamic components of power produced, and the hysteresis effects between periods of active power harvesting and dormancy; the effects of 'on sediment surface' versus 'in sediment' anode deployment have been examined for smaller electrode areas so far. A capacitance model of power consumption and harvesting has been proposed for the marine environment. It is assumed that the primordial benthic microbe laden layer of the earth acts like a giant capacitor. In the microbial fuel cell, this charged benthic layer acts in series with a smaller constant voltage DC power source. This giant benthic capacitance is a result of untapped accumulated charge from the microbes while the DC source originates from the real-time production due to the microbes. Finally, the microbial fuel cell is integrated with a power conversion system to intermittently energize a

  5. Programmed Cell Death and Complexity in Microbial Systems.

    PubMed

    Durand, Pierre M; Sym, Stuart; Michod, Richard E

    2016-07-11

    Programmed cell death (PCD) is central to organism development and for a long time was considered a hallmark of multicellularity. Its discovery, therefore, in unicellular organisms presents compelling questions. Why did PCD evolve? What is its ecological effect on communities? To answer these questions, one is compelled to consider the impacts of PCD beyond the cell, for death obviously lowers the fitness of the cell. Here, we examine the ecological effects of PCD in different microbial scenarios and conclude that PCD can increase biological complexity. In mixed microbial communities, the mode of death affects the microenvironment, impacting the interactions between taxa. Where the population comprises groups of relatives, death has a more explicit effect. Death by lysis or other means can be harmful, while PCD can evolve by providing advantages to relatives. The synchronization of death between individuals suggests a group level property is being maintained and the mode of death also appears to have had an impact during the origin of multicellularity. PCD can result in the export of fitness from the cell to the group level via re-usable resources and PCD may also provide a mechanism for how groups beget new groups comprising kin. Furthermore, PCD is a means for solving a central problem of group living - the toxic effects of death - by making resources in dying cells beneficial to others. What emerges from the data reviewed here is that while PCD carries an obvious cost to the cell, it can be a driver of complexity in microbial communities. PMID:27404254

  6. Evaluation of hydrolysis and fermentation rates in microbial fuel cells.

    PubMed

    Velasquez-Orta, Sharon B; Yu, Eileen; Katuri, Krishna P; Head, Ian M; Curtis, Tom P; Scott, Keith

    2011-04-01

    This study determined the influence of substrate degradation on power generation in microbial fuel cells (MFCs) and microbial community selection on the anode. Air cathode MFCs were fed synthetic medium containing different substrates (acetate, glucose and starch) using primary clarifier sewage as source of electroactive bacteria. The complexity of the substrate affected the MFC performance both for power generation and COD removal. Power output decreased with an increase in substrate complexity from 99±2 mWm(-2) for acetate to 4±2 mWm(-2) for starch. The organic matter removal and coulombic efficiency (CE) of MFCs with acetate and glucose (82% of COD removal and 26% CE) were greater than MFCs using starch (60% of COD removal and 19% of CE). The combined hydrolysis-fermentation rate obtained (0.0024 h(-1)) was considerably lower than the fermentation rate (0.018 h(-1)), indicating that hydrolysis of complex compounds limits current output over fermentation. Statistical analysis of microbial community fingerprints, developed on the anode, showed that microbial communities were enriched according to the type of substrate used. Microbial communities producing high power outputs (fed acetate) clustered separately from bacterial communities producing low power outputs (fed complex compounds). PMID:21347728

  7. Simultaneous Alcoholic and Malolactic Fermentations by Saccharomyces cerevisiae and Oenococcus oeni Cells Co-immobilized in Alginate Beads

    PubMed Central

    Bleve, Gianluca; Tufariello, Maria; Vetrano, Cosimo; Mita, Giovanni; Grieco, Francesco

    2016-01-01

    Malolactic fermentation (MLF) usually takes place after the end of alcoholic fermentation (AF). However, the inoculation of lactic acid bacteria together with yeast starter cultures is a promising system to enhance the quality and safety of wine. In recent years, the use of immobilized cell systems has been investigated, with interesting results, for the production of different fermented foods and beverages. In this study we have carried out the simultaneous immobilization of Saccharomyces cerevisiae and Oenococcus oeni in alginate beads and used them in microvinifications tests to produce Negroamaro wine. The process was monitored by chemical and sensorial analyses and dominance of starters and cell leaking from beads were also checked. Co-immobilization of S. cerevisiae and O. oeni allowed to perform an efficient fermentation process, producing low volatile acidity levels and ethanol and glycerol concentrations comparable with those obtained by cell sequential inoculum and co-inoculum of yeast and bacteria cells in free form. More importantly, co-immobilization strategy produced a significant decrease of the time requested to complete AF and MLF. The immobilized cells could be efficiently reused for the wine fermentation at least three times without any apparent loss of cell metabolic activities. This integrated biocatalytic system is able to perform simultaneously AF and MLF, producing wines similar in organoleptic traits in comparison with wines fermented following traditional sequential AF and MLF with free cell starters. The immobilized-cell system, that we here describe for the first time in our knowledge, offers many advantages over conventional free cell fermentations, including: (i) elimination of non-productive cell growth phases; (ii) feasibility of continuous processing; (iii) re-use of the biocatalyst. PMID:27379072

  8. Identifying the microbial communities and operational conditions for optimized wastewater treatment in microbial fuel cells.

    PubMed

    Ishii, Shun'ichi; Suzuki, Shino; Norden-Krichmar, Trina M; Wu, Angela; Yamanaka, Yuko; Nealson, Kenneth H; Bretschger, Orianna

    2013-12-01

    Microbial fuel cells (MFCs) are devices that exploit microorganisms as "biocatalysts" to recover energy from organic matter in the form of electricity. MFCs have been explored as possible energy neutral wastewater treatment systems; however, fundamental knowledge is still required about how MFC-associated microbial communities are affected by different operational conditions and can be optimized for accelerated wastewater treatment rates. In this study, we explored how electricity-generating microbial biofilms were established at MFC anodes and responded to three different operational conditions during wastewater treatment: 1) MFC operation using a 750 Ω external resistor (0.3 mA current production); 2) set-potential (SP) operation with the anode electrode potentiostatically controlled to +100 mV vs SHE (4.0 mA current production); and 3) open circuit (OC) operation (zero current generation). For all reactors, primary clarifier effluent collected from a municipal wastewater plant was used as the sole carbon and microbial source. Batch operation demonstrated nearly complete organic matter consumption after a residence time of 8-12 days for the MFC condition, 4-6 days for the SP condition, and 15-20 days for the OC condition. These results indicate that higher current generation accelerates organic matter degradation during MFC wastewater treatment. The microbial community analysis was conducted for the three reactors using 16S rRNA gene sequencing. Although the inoculated wastewater was dominated by members of Epsilonproteobacteria, Gammaproteobacteria, and Bacteroidetes species, the electricity-generating biofilms in MFC and SP reactors were dominated by Deltaproteobacteria and Bacteroidetes. Within Deltaproteobacteria, phylotypes classified to family Desulfobulbaceae and Geobacteraceae increased significantly under the SP condition with higher current generation; however those phylotypes were not found in the OC reactor. These analyses suggest that species

  9. Cell Walls of Saccharomyces cerevisiae Differentially Modulated Innate Immunity and Glucose Metabolism during Late Systemic Inflammation

    PubMed Central

    Baurhoo, Bushansingh; Ferket, Peter; Ashwell, Chris M.; de Oliviera, Jean; Zhao, Xin

    2012-01-01

    Background Salmonella causes acute systemic inflammation by using its virulence factors to invade the intestinal epithelium. But, prolonged inflammation may provoke severe body catabolism and immunological diseases. Salmonella has become more life-threatening due to emergence of multiple-antibiotic resistant strains. Mannose-rich oligosaccharides (MOS) from cells walls of Saccharomyces cerevisiae have shown to bind mannose-specific lectin of Gram-negative bacteria including Salmonella, and prevent their adherence to intestinal epithelial cells. However, whether MOS may potentially mitigate systemic inflammation is not investigated yet. Moreover, molecular events underlying innate immune responses and metabolic activities during late inflammation, in presence or absence of MOS, are unknown. Methods and Principal Findings Using a Salmonella LPS-induced systemic inflammation chicken model and microarray analysis, we investigated the effects of MOS and virginiamycin (VIRG, a sub-therapeutic antibiotic) on innate immunity and glucose metabolism during late inflammation. Here, we demonstrate that MOS and VIRG modulated innate immunity and metabolic genes differently. Innate immune responses were principally mediated by intestinal IL-3, but not TNF-α, IL-1 or IL-6, whereas glucose mobilization occurred through intestinal gluconeogenesis only. MOS inherently induced IL-3 expression in control hosts. Consequent to LPS challenge, IL-3 induction in VIRG hosts but not differentially expressed in MOS hosts revealed that MOS counteracted LPS's detrimental inflammatory effects. Metabolic pathways are built to elucidate the mechanisms by which VIRG host's higher energy requirements were met: including gene up-regulations for intestinal gluconeogenesis (PEPCK) and liver glycolysis (ENO2), and intriguingly liver fatty acid synthesis through ATP citrate synthase (CS) down-regulation and ATP citrate lyase (ACLY) and malic enzyme (ME) up-regulations. However, MOS host's lower energy

  10. Efficacy of beer fermentation residue containing Saccharomyces cerevisiae cells for ameliorating aflatoxicosis in broilers.

    PubMed

    Bovo, F; Franco, L T; Kobashigawa, E; Rottinghaus, G E; Ledoux, D R; Oliveira, C A F

    2015-05-01

    This study aimed to determine the aflatoxin B1 (AFB1) binding capacity of a beer fermentation residue (BFR) containing Saccharomyces cerevisiae cells, and the efficacy of BFR to ameliorate the toxic effects of AFB1 on performance, serum biochemistry, and histology of broilers. The BFR was collected from a microbrewery, and the yeast cells were counted, dried, and milled before it was used in the study. In vitro evaluation of the BFR was conducted using different concentrations of AFB1 (2.0, 4.0, 8.0, 16.0, and 32.0 μg AFB1/mL) and 100 mg/10 mL of BFR at pH 3.0 or 6.0. Two hundred 1-day-old male broilers (Ross 308) were assigned to chick batteries and allowed ad libitum access to feed and water. A completely randomized design was used with 5 replicate pens of 5 chicks assigned to each of 4 dietary treatments from hatch to 21 d, which included: 1) basal diet (BD), with no BFR or AFB1; 2) BD supplemented with 1% BFR; 3) BD supplemented with 2 mg AFB1/kg of feed; and 4) BD supplemented with 2 mg AFB1/kg feed and 1% BFR. Performance variables were determined weekly, while serum analyses were performed on d 14 and 21. At the end of the study, chicks were anesthetized with carbon dioxide, euthanized by cervical dislocation, and the kidney, liver, and bursa of Fabricius were removed for determination of relative weights, and for histological evaluation. In vitro assays showed that the higher the initial AFB1 concentration in solution, the greater the AFB1 amount adsorbed by BFR at both pHs tested. Feed intake, BW gain, and concentrations of albumin, total protein, and globulin increased (P < 0.05) in broilers fed BFR+AFB1 (Diet 4), when compared to the birds receiving only AFB1 (Diet 2). Although BFR was not able to reduce or prevent the effects of AFB1 on relative weights of kidneys and liver, it reduced the severity of histological changes in the liver and kidney caused by AFB1. PMID:25743420

  11. Interactions of Condensed Tannins with Saccharomyces cerevisiae Yeast Cells and Cell Walls: Tannin Location by Microscopy.

    PubMed

    Mekoue Nguela, Julie; Vernhet, Aude; Sieczkowski, Nathalie; Brillouet, Jean-Marc

    2015-09-01

    Interactions between grape tannins/red wine polyphenols and yeast cells/cell walls was previously studied within the framework of red wine aging and the use of yeast-derived products as an alternative to aging on lees. Results evidenced a quite different behavior between whole cells (biomass grown to elaborate yeast-derived products, inactivated yeast, and yeast inactivated after autolysis) and yeast cell walls (obtained from mechanical disruption of the biomass). Briefly, whole cells exhibited a high capacity to irreversibly adsorb grape and wine tannins, whereas only weak interactions were observed for cell walls. This last point was quite unexpected considering the literature and called into question the real role of cell walls in yeasts' ability to fix tannins. In the present work, tannin location after interactions between grape and wine tannins and yeast cells and cell walls was studied by means of transmission electron microscopy, light epifluorescence, and confocal microscopy. Microscopy observations evidenced that if tannins interact with cell walls, and especially cell wall mannoproteins, they also diffuse freely through the walls of dead cells to interact with their plasma membrane and cytoplasmic components. PMID:26223789

  12. Development of microbial cell factories for bio-refinery through synthetic bioengineering.

    PubMed

    Kondo, Akihiko; Ishii, Jun; Hara, Kiyotaka Y; Hasunuma, Tomohisa; Matsuda, Fumio

    2013-01-20

    Synthetic bioengineering is a strategy for developing useful microbial strains with innovative biological functions. Novel functions are designed and synthesized in host microbes with the aid of advanced technologies for computer simulations of cellular processes and the system-wide manipulation of host genomes. Here, we review the current status and future prospects of synthetic bioengineering in the yeast Saccharomyces cerevisiae for bio-refinery processes to produce various commodity chemicals from lignocellulosic biomass. Previous studies to improve assimilation of xylose and production of glutathione and butanol suggest a fixed pattern of problems that need to be solved, and as a crucial step, we now need to identify promising targets for further engineering of yeast metabolism. Metabolic simulation, transcriptomics, and metabolomics are useful emerging technologies for achieving this goal, making it possible to optimize metabolic pathways. Furthermore, novel genes responsible for target production can be found by analyzing large-scale data. Fine-tuning of enzyme activities is essential in the latter stage of strain development, but it requires detailed modeling of yeast metabolic functions. Recombinant technologies and genetic engineering are crucial for implementing metabolic designs into microbes. In addition to conventional gene manipulation techniques, advanced methods, such as multicistronic expression systems, marker-recycle gene deletion, protein engineering, cell surface display, genome editing, and synthesis of very long DNA fragments, will facilitate advances in synthetic bioengineering. PMID:22728424

  13. Microbial fuel cell (MFC) for bioelectricity generation from organic wastes.

    PubMed

    Moqsud, M Azizul; Omine, Kiyoshi; Yasufuku, Noriyuki; Hyodo, Masayuki; Nakata, Yukio

    2013-11-01

    Microbial fuel cells (MFCs) have gained a lot of attention recently as a mode of converting organic matter into electricity. In this study, a compost-based microbial fuel cell that generates bioelectricity by biodegradation of organic matter is developed. Grass cuttings, along with leaf mold, rice bran, oil cake (from mustard plants) and chicken droppings (waste from chickens) were used as organic waste. The electric properties of the MFC under anaerobic fermentation condition were investigated along with the influence of different types of membranes, the mixing of fly ash, and different types of electrode materials. It is observed that the maximum voltage was increased by mixing fly ash. Cellophane showed the highest value of voltage (around 350mV). Bamboo charcoal is good for anode material; however carbon fiber is better for the cathode material in terms of optimization of power generated. This developed MFC is a simple cell to generate electricity from organic waste. PMID:23962448

  14. Microbial community structure accompanied with electricity production in a constructed wetland plant microbial fuel cell.

    PubMed

    Lu, Lu; Xing, Defeng; Ren, Zhiyong Jason

    2015-11-01

    This study reveals the complex structure of bacterial and archaeal communities associated with a Canna indica plant microbial fuel cell (PMFC) and its electricity production. The PMFC produced a maximum current of 105 mA/m(2) by utilizing rhizodeposits as the sole electron donor without any external nutrient or buffer supplements, which demonstrates the feasibility of PMFCs in practical oligotrophic conditions with low solution conductivity. The microbial diversity was significantly higher in the PMFC than non-plant controls or sediment-only controls, and pyrosequencing and clone library reveal that rhizodeposits conversion to current were carried out by syntrophic interactions between fermentative bacteria (e.g., Anaerolineaceae) and electrochemically active bacteria (e.g., Geobacter). Denitrifying bacteria and acetotrophic methanogens play a minor role in organics degradation, but abundant hydrogenotrophic methanogens and thermophilic archaea are likely main electron donor competitors. PMID:26066972

  15. Display of phytase on the cell surface of Saccharomyces cerevisiae to degrade phytate phosphorus and improve bioethanol production.

    PubMed

    Chen, Xianzhong; Xiao, Yan; Shen, Wei; Govender, Algasan; Zhang, Liang; Fan, You; Wang, Zhengxiang

    2016-03-01

    Currently, development of biofuels as an alternative fuel has gained much attention due to resource and environmental challenges. Bioethanol is one of most important and dominant biofuels, and production using corn or cassava as raw materials has become a prominent technology. However, phytate contained in the raw material not only decreases the efficiency of ethanol production, but also leads to an increase in the discharge of phosphorus, thus impacting on the environment. In this study, to decrease phytate and its phosphorus content in an ethanol fermentation process, Saccharomyces cerevisiae was engineered through a surface-displaying system utilizing the C-terminal half of the yeast α-agglutinin protein. The recombinant yeast strain, PHY, was constructed by successfully displaying phytase on the surface of cells, and enzyme activity reached 6.4 U/g wet biomass weight. Ethanol productions using various strains were compared, and the results demonstrated that the specific growth rate and average fermentation rate of the PHY strain were higher 20 and 18 %, respectively, compared to the control strain S. cerevisiae CICIMY0086, in a 5-L bioreactor process by simultaneous saccharification and fermentation. More importantly, the phytate phosphorus concentration decreased by 89.8 % and free phosphorus concentration increased by 142.9 % in dry vinasse compared to the control in a 5-L bioreactor. In summary, we constructed a recombinant S. cerevisiae strain displaying phytase on the cell surface, which could improve ethanol production performance and effectively reduce the discharge of phosphorus. The strain reported here represents a useful novel engineering platform for developing an environment-friendly system for bioethanol production from a corn substrate. PMID:26610799

  16. Microbial cell retention in a melting High Arctic snowpack, Svalbard

    NASA Astrophysics Data System (ADS)

    Zarsky, Jakub; Björkman, Mats; Kühnel, Rafael; Hell, Katherina; Hodson, Andy; Sattler, Birgit; Psenner, Roland

    2014-05-01

    Introduction The melting snow pack represents a highly dynamic system not only for chemical compounds but also for bacterial cells. Microbial activity was found at subzero temperatures in ice veins when liquid water persists due to high concentration of ions on the surface of snow crystals and brine channels between large ice crystals in ice. Several observations also suggest microbial activity under subzero temperatures in seasonal snow. Even with regard to the spatial and temporal relevance of snow ecosystems, microbial activity in such an extreme habitat represents a relatively small proportion in the carbon flux of the global ecosystem and even of the glacial ecosystems specifically. On the other hand, it represents a remarkable piece of mosaic of the microbial activity in glacial ecosystems because the snow pack represents the first contact between the atmosphere and cryosphere. This topic also embodies vital crossovers to biogeochemistry and ecotoxicology, offering a quantitative view of utilization of various substrates relevant for downstream ecosystems. Here we present our study of the dynamics of both solvents and cells suspended in meltwater of the melting snowpack on a high arctic glacier to demonstrate the spatio-temporal constraint of interaction between solvent and bacterial cells in this environment. Method We used 6 lysimeters inserted into the bottom of the snowpack to collect replicated samples of melt water before it comes into contact with basal ice or slush layer at the base of the snow pack. The sampling site was chosen at Midre Lovénbreen (Svalbard, Kongsfjorden, MLB stake 6) where the snow pack showed melting on the surface but the basal ice was still dry. Sampling was conducted in June 2010 for a period of 10 days once per day and the snow profile was sampled according to distinguished layers in the profile at the beginning of the field mission and as bulk at its end. The height of snow above the lysimeters dropped from the initial 74 cm

  17. Isolation of mannan-protein complexes from viable cells of Saccharomyces cerevisiae X2180-1A wild type and Saccharomyces cerevisiae X2180-1 A-5 mutant strains by the action of Zymolyase-60,000.

    PubMed Central

    Shibata, N; Mizugami, K; Takano, K; Suzuki, S

    1983-01-01

    The viable whole cells of Saccharomyces cerevisiae X2180-1A wild type and its mannan mutant strain S. cerevisiae X2180-1A-5, were treated with an Arthrobacter sp. beta-1,3-glucanase in the presence of a serine protease inhibitor, phenyl-methylsulfonyl fluoride. Fractionation of the solubilized materials of each strain with Cetavlon (cetyltrimethylammonium bromide) yielded one mannan-protein complex. Molecular weights of these complexes were almost the same as that of the mannoprotein of the mutant strain prepared by Nakajima and Ballou, which had a molecular weight of 133,000 and were approximately three times larger than those of the mannans isolated from the same cells by hot-water extraction. Each mannan-protein complex contained up to 2% glucose residue, which was not removed by specific precipitation with anti-mannan sera or by affinity chromatography on a column of concanavalin A-Sepharose. Treatment of these complexes with alkaline NaBH4 produced peptide-free mannan containing small amounts of glucose nearly identical to those of the parent complexes. The above findings provide evidence that the glucose residues exist in a covalently linked form to the mannan moiety. Fractionation of the mannan-protein complex of the S. cerevisiae wild-type strain by DEAE-Sephadex chromatography yielded five subfractions of different phosphate content, indicating that these highly intact mannan-protein complexes were of heterogeneous material consisting of many molecular species of different phosphate content. PMID:6355061

  18. Enhanced microbial reduction of vanadium (V) in groundwater with bioelectricity from microbial fuel cells

    NASA Astrophysics Data System (ADS)

    Hao, Liting; Zhang, Baogang; Tian, Caixing; Liu, Ye; Shi, Chunhong; Cheng, Ming; Feng, Chuanping

    2015-08-01

    Bioelectricity generated from the microbial fuel cell (MFC) is applied to the bioelectrical reactor (BER) directly to enhance microbial reduction of vanadium (V) (V(V)) in groundwater. With the maximum power density of 543.4 mW m-2 from the MFC, V(V) removal is accelerated with efficiency of 93.6% during 12 h operation. Higher applied voltage can facilitate this process. V(V) removals decrease with the increase of initial V(V) concentration, while extra addition of chemical oxygen demand (COD) has little effect on performance improvement. Microbial V(V) reduction is enhanced and then suppressed with the increase of conductivity. High-throughput 16S rRNA gene pyrosequencing analysis implies the accumulated Enterobacter and Lactococcus reduce V(V) with products from fermentative microorganisms such as Macellibacteroides. The presentation of electrochemically active bacteria as Enterobacter promotes electron transfers. This study indicates that application of bioelectricity from MFCs is a promising strategy to improve the efficiency of in-situ bioremediation of V(V) polluted groundwater.

  19. Effect of electricity on microbial community of microbial fuel cell simultaneously treating sulfide and nitrate

    NASA Astrophysics Data System (ADS)

    Cai, Jing; Zheng, Ping; Xing, Yajuan; Qaisar, Mahmood

    2015-05-01

    The effect of electric current on microbial community is explored in Microbial Fuel Cells (MFCs) simultaneously treating sulfide and nitrate. The MFCs are operated under four different conditions which exhibited different characteristics of electricity generation. In batch mode, MFCs generate intermittently high current pulses in the beginning, and the current density is instable subsequently, while the current density of MFCs in continuous mode is relatively stable. All operational parameters show good capacity for substrate removal, and nitrogen and sulfate were the main reaction products. Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) analysis is employed to obtain profiles of the bacterial communities present in inoculum and suspension of four MFCs. Based on the community diversity indices and Spearman correlation analyses, significant correlation exists between Richness of the community of anode chamber and the electricity generated, while no strong correlation is evident between other indexes (Shannon index, Simpson index and Equitability index) and the electricity. Additionally, the results of Principal Component Analysis (PCA) suggest that MFCs suffering from current shock have similar suspension communities, while the others have diverse microbial communities.

  20. Microbial community structures differentiated in a single-chamber air-cathode microbial fuel cell fueled with rice straw hydrolysate

    PubMed Central

    2014-01-01

    Background The microbial fuel cell represents a novel technology to simultaneously generate electric power and treat wastewater. Both pure organic matter and real wastewater can be used as fuel to generate electric power and the substrate type can influence the microbial community structure. In the present study, rice straw, an important feedstock source in the world, was used as fuel after pretreatment with diluted acid method for a microbial fuel cell to obtain electric power. Moreover, the microbial community structures of anodic and cathodic biofilm and planktonic culturewere analyzed and compared to reveal the effect of niche on microbial community structure. Results The microbial fuel cell produced a maximum power density of 137.6 ± 15.5 mW/m2 at a COD concentration of 400 mg/L, which was further increased to 293.33 ± 7.89 mW/m2 through adjusting the electrolyte conductivity from 5.6 mS/cm to 17 mS/cm. Microbial community analysis showed reduction of the microbial diversities of the anodic biofilm and planktonic culture, whereas diversity of the cathodic biofilm was increased. Planktonic microbial communities were clustered closer to the anodic microbial communities compared to the cathodic biofilm. The differentiation in microbial community structure of the samples was caused by minor portion of the genus. The three samples shared the same predominant phylum of Proteobacteria. The abundance of exoelectrogenic genus was increased with Desulfobulbus as the shared most abundant genus; while the most abundant exoelectrogenic genus of Clostridium in the inoculum was reduced. Sulfate reducing bacteria accounted for large relative abundance in all the samples, whereas the relative abundance varied in different samples. Conclusion The results demonstrated that rice straw hydrolysate can be used as fuel for microbial fuel cells; microbial community structure differentiated depending on niches after microbial fuel cell operation; exoelectrogens were

  1. The linkage of (1-3)-beta-glucan to chitin during cell wall assembly in Saccharomyces cerevisiae.

    PubMed

    Hartland, R P; Vermeulen, C A; Klis, F M; Sietsma, J H; Wessels, J G

    1994-12-01

    Pulse-chase experiments with [14C]glucose demonstrated that in the cell wall of wild-type Saccharomyces cerevisiae alkali-soluble (1-3)-beta-glucan serves as a precursor for alkali-insoluble (1-3)-beta-glucan. The following observations support the notion that the insolubilization of the glucan is caused by linkage to chitin: (i) degradation of chitin by chitinase completely dissolved the glucan, and (ii) disruption of the gene for chitin synthase 3 prevented the formation of alkali-insoluble glucan. These cells, unable to form a glucan-chitin complex, were highly vulnerable to hypo-osmotic shock indicating that the linkage of the two polymers significantly contributes to the mechanical strength of the cell wall. Conversion of alkali-soluble glucan into alkali-insoluble glucan occurred both early and late during budding and also in the ts-mutant cdc24-1 in the absence of bud formation. PMID:7725794

  2. Determination of maintenance coefficients of Saccharomyces cerevisiae cultures with cell recycle by cross-flow membrane filtration.

    PubMed

    Uribelarrea, J L; Winter, J; Goma, G; Pareilleux, A

    1990-01-20

    The fermentation of glucose by a strain of Saccharomyces cerevisiae was studied in a continuous single-stage process with recycle of the cells via cross-flow micro-filtration membranes. Operating conditions were selected such that the culture was not carbon limited and inhibition by ethanol and cell death were minimized.Steady states were obtained for various biomass bleeding rates, i.e., various specific growth rates. From the experimental data, the stoichiometry of the simultaneous reactions, cell growth, ethanol production and maintenance were established using mass and degree of reduction balance relative to substrates (carbon source and oxygen) and products (biomass, ethanol, carbon dioxide etc.), and the growth parameters, yields, and maintenance cofficients were determined. It was shown that the oxygen consumption was not linked to the kinetics of the fermentation. The calculated growth constants were discussed and compared to the currently reported values. PMID:18592509

  3. Recent developments in microbial fuel cell technologies for sustainable bioenergy.

    PubMed

    Watanabe, Kazuya

    2008-12-01

    Microbial fuel cells (MFCs) are devices that exploit microbial catabolic activities to generate electricity from a variety of materials, including complex organic waste and renewable biomass. These sources provide MFCs with a great advantage over chemical fuel cells that can utilize only purified reactive fuels (e.g., hydrogen). A developing primary application of MFCs is its use in the production of sustainable bioenergy, e.g., organic waste treatment coupled with electricity generation, although further technical developments are necessary for its practical use. In this article, recent advances in MFC technologies that can become fundamentals for future practical MFC developments are summarized. Results of recent studies suggest that MFCs will be of practical use in the near future and will become a preferred option among sustainable bioenergy processes. PMID:19134546

  4. Produced Water Treatment Using Microbial Fuel Cell Technology

    SciTech Connect

    Borole, A. P.; Campbell, R.

    2011-05-20

    ORNL has developed a treatment for produced water using a combination of microbial fuel cells and electrosorption. A collaboration between Campbell Applied Physics and ORNL was initiated to further investigate development of the technology and apply it to treatment of field produced water. The project successfully demonstrated the potential of microbial fuel cells to generate electricity from organics in produced water. A steady voltage was continuously generated for several days using the system developed in this study. In addition to the extraction of electrical energy from the organic contaminants, use of the energy at the representative voltage was demonstrated for salts removal or desalination of the produced water. Thus, the technology has potential to remove organic as well as ionic contaminants with minimal energy input using this technology. This is a novel energy-efficient method to treat produced water. Funding to test the technology at larger scale is being pursued to enable application development.

  5. New insights in Microbial Fuel Cells: novel solid phase anolyte.

    PubMed

    Tommasi, Tonia; Salvador, Gian Paolo; Quaglio, Marzia

    2016-01-01

    For the development of long lasting portable microbial fuel cells (MFCs) new strategies are necessary to overcome critical issues such as hydraulic pump system and the biochemical substrate retrieval overtime to sustain bacteria metabolism. The present work proposes the use of a synthetic solid anolyte (SSA), constituted by agar, carbonaceous and nitrogen sources dissolved into diluted seawater. Results of a month-test showed the potential of the new SSA-MFC as a long lasting low energy consuming system. PMID:27375205

  6. Enzyme Amplified Detection of Microbial Cell Wall Components

    NASA Technical Reports Server (NTRS)

    Wainwright, Norman R.

    2004-01-01

    This proposal is MBL's portion of NASA's Johnson Space Center's Astrobiology Center led by Principal Investigator, Dr. David McKay, entitled: 'Institute for the Study of Biomarkers in Astromaterials.' Dr. Norman Wainwright is the principal investigator at MBL and is responsible for developing methods to detect trace quantities of microbial cell wall chemicals using the enzyme amplification system of Limulus polyphemus and other related methods.

  7. New insights in Microbial Fuel Cells: novel solid phase anolyte

    PubMed Central

    Tommasi, Tonia; Salvador, Gian Paolo; Quaglio, Marzia

    2016-01-01

    For the development of long lasting portable microbial fuel cells (MFCs) new strategies are necessary to overcome critical issues such as hydraulic pump system and the biochemical substrate retrieval overtime to sustain bacteria metabolism. The present work proposes the use of a synthetic solid anolyte (SSA), constituted by agar, carbonaceous and nitrogen sources dissolved into diluted seawater. Results of a month-test showed the potential of the new SSA-MFC as a long lasting low energy consuming system. PMID:27375205

  8. New insights in Microbial Fuel Cells: novel solid phase anolyte

    NASA Astrophysics Data System (ADS)

    Tommasi, Tonia; Salvador, Gian Paolo; Quaglio, Marzia

    2016-07-01

    For the development of long lasting portable microbial fuel cells (MFCs) new strategies are necessary to overcome critical issues such as hydraulic pump system and the biochemical substrate retrieval overtime to sustain bacteria metabolism. The present work proposes the use of a synthetic solid anolyte (SSA), constituted by agar, carbonaceous and nitrogen sources dissolved into diluted seawater. Results of a month-test showed the potential of the new SSA-MFC as a long lasting low energy consuming system.

  9. A computational model for biofilm-based microbial fuel cells.

    PubMed

    Picioreanu, Cristian; Head, Ian M; Katuri, Krishna P; van Loosdrecht, Mark C M; Scott, Keith

    2007-07-01

    This study describes and evaluates a computational model for microbial fuel cells (MFCs) based on redox mediators with several populations of suspended and attached biofilm microorganisms, and multiple dissolved chemical species. A number of biological, chemical and electrochemical reactions can occur in the bulk liquid, in the biofilm and at the electrode surface. The evolution in time of important MFC parameters (current, charge, voltage and power production, consumption of substrates, suspended and attached biomass growth) has been simulated under several operational conditions. Model calculations evaluated the effect of different substrate utilization yields, standard potential of the redox mediator, ratio of suspended to biofilm cells, initial substrate and mediator concentrations, mediator diffusivity, mass transfer boundary layer, external load resistance, endogenous metabolism, repeated substrate additions and competition between different microbial groups in the biofilm. Two- and three-dimensional model simulations revealed the heterogeneous current distribution over the planar anode surface for younger and patchy biofilms, but becoming uniform in older and more homogeneous biofilms. For uniformly flat biofilms one-dimensional models should give sufficiently accurate descriptions of produced currents. Voltage- and power-current characteristics can also be calculated at different moments in time to evaluate the limiting regime in which the MFC operates. Finally, the model predictions are tested with previously reported experimental data obtained in a batch MFC with a Geobacter biofilm fed with acetate. The potential of the general modeling framework presented here is in the understanding and design of more complex cases of wastewater-fed microbial fuel cells. PMID:17537478

  10. In Situ fuel processing in a microbial fuel cell.

    PubMed

    Bahartan, Karnit; Amir, Liron; Israel, Alvaro; Lichtenstein, Rachel G; Alfonta, Lital

    2012-09-01

    A microbial fuel cell (MFC) was designed in which fuel is generated in the cell by the enzyme glucoamylase, which is displayed on the surface of yeast. The enzyme digests starch specifically into monomeric glucose units and as a consequence enables further glucose oxidation by microorganisms present in the MFC anode. The oxidative enzyme glucose oxidase was coupled to the glucoamylase digestive enzyme. When both enzymes were displayed on the surface of yeast cells in a mixed culture, superior fuel-cell performance was observed in comparison with other combinations of yeast cells, unmodified yeast, or pure enzymes. The feasibility of the use of the green macroalgae Ulva lactuca in such a genetically modified MFC was also demonstrated. Herein, we report the performance of such fuel cells as a proof of concept for the enzymatic digestion of complex organic fuels in the anode of MFCs to render the fuel more available to microorganisms. PMID:22833422

  11. Recent Advances in Genetic Technique of Microbial Report Cells and Their Applications in Cell Arrays

    PubMed Central

    Kim, Do Hyun; Kim, Moon Il; Park, Hyun Gyu

    2015-01-01

    Microbial cell arrays have attracted consistent attention for their ability to provide unique global data on target analytes at low cost, their capacity for readily detectable and robust cell growth in diverse environments, their high degree of convenience, and their capacity for multiplexing via incorporation of molecularly tailored reporter cells. To highlight recent progress in the field of microbial cell arrays, this review discusses research on genetic engineering of reporter cells, technologies for patterning live cells on solid surfaces, cellular immobilization in different polymers, and studies on their application in environmental monitoring, disease diagnostics, and other related fields. On the basis of these results, we discuss current challenges and future prospects for novel microbial cell arrays, which show promise for use as potent tools for unraveling complex biological processes. PMID:26436087

  12. Proline accumulation protects Saccharomyces cerevisiae cells in stationary phase from ethanol stress by reducing reactive oxygen species levels.

    PubMed

    Takagi, Hiroshi; Taguchi, Junpei; Kaino, Tomohiro

    2016-08-01

    During fermentation processes, Saccharomyces cerevisiae cells are exposed to multiple stresses, including a high concentration of ethanol that represents toxicity through intracellular reactive oxygen species (ROS) generation. We previously reported that proline protected yeast cells from damage caused by various stresses, such as freezing and ethanol. As an anti-oxidant, proline is suggested to scavenge intracellular ROS. In this study, we examined the role of intracellular proline during ethanol treatment in S. cerevisiae strains that accumulate different concentrations of proline. When cultured in YPD medium, there was a significant accumulation of proline in the put1 mutant strain, which is deficient in proline oxidase, in the stationary phase. Expression of the mutant PRO1 gene, which encodes the γ-glutamyl kinase variant (Asp154Asn or Ile150Thr) with desensitization to feedback inhibition by proline in the put1 mutant strain, showed a prominent increase in proline content as compared with that of the wild-type strain. The oxidation level was clearly increased in wild-type cells after exposure to ethanol, indicating that the generation of ROS occurred. Interestingly, proline accumulation significantly reduces the ROS level and increases the survival rate of yeast cells in the stationary phase under ethanol stress conditions. However, there was not a clear correlation between proline content and survival rate in yeast cells. An appropriate level of intracellular proline in yeast might be important for its stress-protective effect. Hence, the engineering of proline metabolism could be promising for breeding stress-tolerant industrial yeast strains. Copyright © 2016 John Wiley & Sons, Ltd. PMID:26833688

  13. Mitochondrial Function in Cell Wall Glycoprotein Synthesis in Saccharomyces cerevisiae NCYC 625 (Wild Type) and [rho0] Mutants

    PubMed Central

    Iung, Annie Rakotoarivony; Coulon, Joël; Kiss, Ferenc; Ekome, Jacques Ngondi; Vallner, Judit; Bonaly, Roger

    1999-01-01

    We studied phosphopeptidomannans (PPMs) of two Saccharomyces cerevisiae NCYC 625 strains (S. diastaticus): a wild type strain grown aerobically, anaerobically, and in the presence of antimycin and a [rho0] mutant grown aerobically and anaerobically. The aerobic wild-type cultures were highly flocculent, but all others were weakly flocculent. Ligands implicated in flocculation of mutants or antimycin-treated cells were not aggregated as much by concanavalin A as were those of the wild type. The [rho0] mutants and antimycin-treated cells differ from the wild type in PPM composition and invertase, acid phosphatase, and glucoamylase activities. PPMs extracted from different cells differ in the protein but not in the glycosidic moiety. The PPMs were less stable in mitochondrion-deficient cells than in wild-type cells grown aerobically, and this difference may be attributable to defective mitochondrial function during cell wall synthesis. The reduced flocculation of cells grown in the presence of antimycin, under anaerobiosis, or carrying a [rho0] mutation may be the consequence of alterations of PPM structures which are the ligands of lectins, both involved in this cell-cell recognition phenomenon. These respiratory chain alterations also affect peripheral, biologically active glycoproteins such as extracellular enzymes and peripheral PPMs. PMID:10583995

  14. Sensitivity of Saccharomyces cerevisiae to the cell-penetrating antifungal peptide PAF26 correlates with endogenous nitric oxide (NO) production.

    PubMed

    Carmona, Lourdes; Gandía, Mónica; López-García, Belén; Marcos, Jose F

    2012-01-01

    PAF26 is a synthetic fungicidal hexapeptide with cell-penetration properties and non-lytic mode of action. We demonstrate herein the endogenous accumulation of reactive oxygen species (ROS) and nitric oxide (NO) in the model fungus Saccharomyces cerevisiae treated with PAF26. However, the S. cerevisiae deletion mutant of YAP1 - the major inductor of defense to oxidative stress - did not show high sensitivity to PAF26 but rather increased resistance, and its ROS accumulation did not differ from that of the parental strain. Cross-protection experiments suggest that the oxidant H(2)O(2) and PAF26 kill yeast through different pathways. Overall, the data indicate that ROS are not the primary antifungal mechanism of the peptide. On the contrary, the PAF26-induced intracellular production of NO was blocked in two distinct resistant mutants: the above mentioned Δyap1, which had the induction of NO disrupted, and the previously reported Δarg1 from the biosynthetic pathway of arginine, which has reduced basal NO levels. The NO synthase inhibitor l-NAME partially restored yeast growth in the presence of PAF26. These findings correlate antifungal activity of PAF26 with NO production and provide a plausible explanation for the resistance phenotype of Δarg1 through its involvement in NO biosynthesis. PMID:22120633

  15. alpha-Factor-mediatd modification of a 32P-labeled protein by MATa cells of Saccharomyces cerevisiae.

    PubMed

    Finkelstein, D B; McAlister, L

    1981-03-10

    Addition of the polypeptide mating pheromone alpha-factor to haploid MATa cells of Saccharomyces cerevisiae results in the modification of a 32P-labeled protein (P17) with an apparent Mr of 17,000 to a form having an apparent Mr of 17,500 (P17). 32P associated with both P17 and P17 exhibits an unusually rapid rate of turnover. The conversion of P17 to P17 precedes the appearance of morphologically abnormal cells and, in contrast to other responses elicited by this pheromone, this change in apparent molecular weight does not require protein synthesis. Upon removal of alpha-factor, the P17/P17 ratio returns to pretreatment levels. PMID:7007388

  16. Sustainable wastewater treatment: how might microbial fuel cells contribute.

    PubMed

    Oh, Sung T; Kim, Jung Rae; Premier, Giuliano C; Lee, Tae Ho; Kim, Changwon; Sloan, William T

    2010-01-01

    The need for cost-effective low-energy wastewater treatment has never been greater. Clean water for our expanding and predominantly urban global population will be expensive to deliver, eats into our diminishing carbon-based energy reserves and consequently contributes to green house gases in the atmosphere and climate change. Thus every potential cost and energy cutting measure for wastewater treatment should be explored. Microbial fuel cells (MFCs) could potentially yield such savings but, to achieve this, requires significant advances in our understanding in a few critical areas and in our designs of the overall systems. Here we review the research which might accelerate our progress towards sustainable wastewater treatment using MFCs: system control and modelling and the understanding of the ecology of the microbial communities that catalyse the generation of electricity. PMID:20688144

  17. Energy generation by fermentation of glucose in a batch flow microbial fuel cell

    NASA Astrophysics Data System (ADS)

    Badea, Silviu-Laurentiu; Enache, Stanica; Tamaian, Radu; Buga, Mihaela-Ramona; Pirvu, Cristian; Varlam, Mihai

    2016-04-01

    In the last years, microbial fuel cells (MFCs) have emerged like a novel research technologies for production of sustainable and clean electricity energy through bioxidation of organic materials, representing a promising alternative to combustion energy sources. In this study, production of bioelectricity in MFC in batch system (dual chambered MFC) was investigated. A dual chambered MFC from glass was built for this purpose. Saccharomyces cerevisiae as an active biocatalyst was explored for power generation. Graphite plates were used as electrodes and glucose as substrate. Saccharomyces cerevisiae was initially grown on a period of 72h at 30 degree Celsius, on medium of modified Sabouraud liquid medium containing 30 g glucose/L. A volume of inoculated medium (80 mL) was transferred in the anode compartment of MFC together with 20 mL glucose 1M, while neutral red was used as mediator (electron shuttle) in concentration of 200 μM in anaerobic anode chamber. Potassium permanganate (KMnO4) was used as oxidizing agent in the cathode in wide concentration range (400 μM-40 000 μM). Cathodic compartment was loaded initially with 40 mM potassium permanganate, and afterwards was supplied two times more with KMnO4 of the same concentration, in order to maintain MFC functionality. The MFC was operated on a water bath heated by a combined hot-plate magnetic-stirrer device at 30 degree Celsius and mixed at 180 rpm. The maximum open circuit potential (OCV) recorded of about 0.6 V was reached after the 3rd loading with 40 milimolles of potassium permanganate. Using a potentiostat, the polarization curve was recorded by varying the potential between 0.5 V and 0.0 V, while the intensity of current increased from 0.0 to about 1.5 mA respectively, corresponding to an anodic current density of about 0.81 A/m2. In order to optimize the design and performance of the MFC, the goal of the further research is to use variously concentrations of potassium permanganate. Furthermore, a dual

  18. Cell cycle execution point analysis of ORC function and characterization of the checkpoint response to ORC inactivation in Saccharomyces cerevisiae.

    PubMed

    Gibson, Daniel G; Bell, Stephen P; Aparicio, Oscar M

    2006-06-01

    Chromosomal replication initiates through the assembly of a prereplicative complex (pre-RC) at individual replication origins in the G1-phase, followed by activation of these complexes in the S-phase. In Saccharomyces cerevisiae, the origin recognition complex (ORC) binds replication origins throughout the cell cycle and participates in pre-RC assembly. Whether the ORC plays an additional role subsequent to pre-RC assembly in replication initiation or any other essential cell cycle process is not clear. To study the function of the ORC during defined cell cycle periods, we performed cell cycle execution point analyses with strains containing a conditional mutation in the ORC1, ORC2 or ORC5 subunit of ORC. We found that the ORC is essential for replication initiation, but is dispensable for replication elongation or later cell cycle events. Defective initiation in ORC mutant cells results in incomplete replication and mitotic arrest enforced by the DNA damage and spindle assembly checkpoint pathways. The involvement of the spindle assembly checkpoint implies a defect in kinetochore-spindle attachment or sister chromatid cohesion due to incomplete replication and/or DNA damage. Remarkably, under semipermissive conditions for ORC1 function, the spindle checkpoint alone suffices to block proliferation, suggesting this checkpoint is highly sensitive to replication initiation defects. We discuss the potential significance of these overlapping checkpoints and the impact of our findings on previously postulated role(s) of ORCs in other cell cycle functions. PMID:16716188

  19. Engineering microbial fuels cells: recent patents and new directions.

    PubMed

    Biffinger, Justin C; Ringeisen, Bradley R

    2008-01-01

    Fundamental research into how microbes generate electricity within microbial fuel cells (MFCs) has far outweighed the practical application and large scale development of microbial energy harvesting devices. MFCs are considered alternatives to standard commercial polymer electrolyte membrane (PEM) fuel cell technology because the fuel supply does not need to be purified, ambient operating temperatures are maintained with biologically compatible materials, and the biological catalyst is self-regenerating. The generation of electricity during wastewater treatment using MFCs may profoundly affect the approach to anaerobic treatment technologies used in wastewater treatment as a result of developing this energy harvesting technology. However, the materials and engineering designs for MFCs were identical to commercial fuel cells until 2003. Compared to commercial fuel cells, MFCs will remain underdeveloped as long as low power densities are generated from the best systems. The variety of designs for MFCs has expanded rapidly in the last five years in the literature, but the patent protection has lagged behind. This review will cover recent and important patents relating to MFC designs and progress. PMID:19075862

  20. Osteoprotegerin Regulates Pancreatic β-Cell Homeostasis upon Microbial Invasion.

    PubMed

    Kuroda, Yukiko; Maruyama, Kenta; Fujii, Hideki; Sugawara, Isamu; Ko, Shigeru B H; Yasuda, Hisataka; Matsui, Hidenori; Matsuo, Koichi

    2016-01-01

    Osteoprotegerin (OPG), a decoy receptor for receptor activator of NF-κB ligand (RANKL), antagonizes RANKL's osteoclastogenic function in bone. We previously demonstrated that systemic administration of lipopolysaccharide (LPS) to mice elevates OPG levels and reduces RANKL levels in peripheral blood. Here, we show that mice infected with Salmonella, Staphylococcus, Mycobacteria or influenza virus also show elevated serum OPG levels. We then asked whether OPG upregulation following microbial invasion had an effect outside of bone. To do so, we treated mice with LPS and observed OPG production in pancreas, especially in β-cells of pancreatic islets. Insulin release following LPS administration was enhanced in mice lacking OPG, suggesting that OPG inhibits insulin secretion under acute inflammatory conditions. Consistently, treatment of MIN6 pancreatic β-cells with OPG decreased their insulin secretion following glucose stimulation in the presence of LPS. Finally, our findings suggest that LPS-induced OPG upregulation is mediated in part by activator protein (AP)-1 family transcription factors, particularly Fos proteins. Overall, we report that acute microbial infection elevates serum OPG, which maintains β-cell homeostasis by restricting glucose-stimulated insulin secretion, possibly preventing microbe-induced exhaustion of β-cell secretory capacity. PMID:26751951

  1. Design and development of synthetic microbial platform cells for bioenergy

    PubMed Central

    Lee, Sang Jun; Lee, Sang-Jae; Lee, Dong-Woo

    2013-01-01

    The finite reservation of fossil fuels accelerates the necessity of development of renewable energy sources. Recent advances in synthetic biology encompassing systems biology and metabolic engineering enable us to engineer and/or create tailor made microorganisms to produce alternative biofuels for the future bio-era. For the efficient transformation of biomass to bioenergy, microbial cells need to be designed and engineered to maximize the performance of cellular metabolisms for the production of biofuels during energy flow. Toward this end, two different conceptual approaches have been applied for the development of platform cell factories: forward minimization and reverse engineering. From the context of naturally minimized genomes,non-essential energy-consuming pathways and/or related gene clusters could be progressively deleted to optimize cellular energy status for bioenergy production. Alternatively, incorporation of non-indigenous parts and/or modules including biomass-degrading enzymes, carbon uptake transporters, photosynthesis, CO2 fixation, and etc. into chassis microorganisms allows the platform cells to gain novel metabolic functions for bioenergy. This review focuses on the current progress in synthetic biology-aided pathway engineering in microbial cells and discusses its impact on the production of sustainable bioenergy. PMID:23626588

  2. Osteoprotegerin Regulates Pancreatic β-Cell Homeostasis upon Microbial Invasion

    PubMed Central

    Kuroda, Yukiko; Maruyama, Kenta; Fujii, Hideki; Sugawara, Isamu; Ko, Shigeru B. H.; Yasuda, Hisataka; Matsui, Hidenori; Matsuo, Koichi

    2016-01-01

    Osteoprotegerin (OPG), a decoy receptor for receptor activator of NF-κB ligand (RANKL), antagonizes RANKL’s osteoclastogenic function in bone. We previously demonstrated that systemic administration of lipopolysaccharide (LPS) to mice elevates OPG levels and reduces RANKL levels in peripheral blood. Here, we show that mice infected with Salmonella, Staphylococcus, Mycobacteria or influenza virus also show elevated serum OPG levels. We then asked whether OPG upregulation following microbial invasion had an effect outside of bone. To do so, we treated mice with LPS and observed OPG production in pancreas, especially in β-cells of pancreatic islets. Insulin release following LPS administration was enhanced in mice lacking OPG, suggesting that OPG inhibits insulin secretion under acute inflammatory conditions. Consistently, treatment of MIN6 pancreatic β-cells with OPG decreased their insulin secretion following glucose stimulation in the presence of LPS. Finally, our findings suggest that LPS-induced OPG upregulation is mediated in part by activator protein (AP)-1 family transcription factors, particularly Fos proteins. Overall, we report that acute microbial infection elevates serum OPG, which maintains β-cell homeostasis by restricting glucose-stimulated insulin secretion, possibly preventing microbe-induced exhaustion of β-cell secretory capacity. PMID:26751951

  3. Scale-up of sediment microbial fuel cells

    NASA Astrophysics Data System (ADS)

    Ewing, Timothy; Ha, Phuc Thi; Babauta, Jerome T.; Tang, Nghia Trong; Heo, Deukhyoun; Beyenal, Haluk

    2014-12-01

    Sediment microbial fuel cells (SMFCs) are used as renewable power sources to operate remote sensors. However, increasing the electrode surface area results in decreased power density, which demonstrates that SMFCs do not scale up with size. As an alternative to the physical scale-up of SMFCs, we proposed that it is possible to scale up power by using smaller-sized individually operated SMFCs connected to a power management system that electrically isolates the anodes and cathodes. To demonstrate our electronic scale-up approach, we operated one 0.36-m2 SMFC (called a single-equivalent SMFC) and four independent SMFCs of 0.09 m2 each (called scaled-up SMFCs) and managed the power using an innovative custom-developed power management system. We found that the single-equivalent SMFC and the scaled-up SMFCs produced similar power for the first 155 days. However, in the long term (>155 days) our scaled-up SMFCs generated significantly more power than the single-equivalent SMFC (2.33 mW vs. 0.64 mW). Microbial community analysis of the single-equivalent SMFC and the scaled-up SMFCs showed very similar results, demonstrating that the difference in operation mode had no significant effect on the microbial community. When we compared scaled-up SMFCs with parallel SMFCs, we found that the scaled-up SMFCs generated more power. Our novel approach demonstrates that SMFCs can be scaled up electronically.

  4. Comparative analysis of microbial community between different cathode systems of microbial fuel cells for denitrification.

    PubMed

    Li, Chao; Xu, Ming; Lu, Yi; Fang, Fang; Cao, Jiashun

    2016-03-01

    Two types of cathodic biofilm in microbial fuel cells (MFC) were established for comparison on their performance and microbial communities. Complete autotrophic simultaneous nitrification and denitrification (SND) without organics addition was achieved in nitrifying-MFC (N-MFC) with a total nitrogen (TN) removal rate of 0.35 mg/(L·h), which was even higher than that in denitrifying-MFC (D-MFC) at same TN level. Integrated denaturing gradient gel electrophoresis analysis based on both 16S rRNA and nirK genes showed that Alpha-, Gammaproteobacteria were the main denitrifier communities. Some potential autotrophic denitrifying bacteria which can use electrons and reducing power from cathodes, such as Shewanella oneidensis, Shewanella loihica, Pseudomonas aeruginosa, Starkeya novella and Rhodopseudomonas palustris were identified and selectively enriched on cathode biofilms. Further, relative abundance of denitrifying bacteria characterized by nirK/16S ratios was much higher in biofilm than suspended sludge according to real-time polymerase chain reaction. The highest enrichment efficiency for denitrifiers was obtained in N-MFC cathode biofilms, which confirmed autotrophic denitrifying bacteria enrichment is the key factor for a D-MFC system. PMID:26278100

  5. Microbial electricity generation in rice paddy fields: recent advances and perspectives in rhizosphere microbial fuel cells.

    PubMed

    Kouzuma, Atsushi; Kaku, Nobuo; Watanabe, Kazuya

    2014-12-01

    Microbial fuel cells (MFCs) are devices that use living microbes for the conversion of organic matter into electricity. MFC systems can be applied to the generation of electricity at water/sediment interfaces in the environment, such as bay areas, wetlands, and rice paddy fields. Using these systems, electricity generation in paddy fields as high as ∼80 mW m(-2) (based on the projected anode area) has been demonstrated, and evidence suggests that rhizosphere microbes preferentially utilize organic exudates from rice roots for generating electricity. Phylogenetic and metagenomic analyses have been conducted to identify the microbial species and catabolic pathways that are involved in the conversion of root exudates into electricity, suggesting the importance of syntrophic interactions. In parallel, pot cultures of rice and other aquatic plants have been used for rhizosphere MFC experiments under controlled laboratory conditions. The findings from these studies have demonstrated the potential of electricity generation for mitigating methane emission from the rhizosphere. Notably, however, the presence of large amounts of organics in the rhizosphere drastically reduces the effect of electricity generation on methane production. Further studies are necessary to evaluate the potential of these systems for mitigating methane emission from rice paddy fields. We suggest that paddy-field MFCs represent a promising approach for harvesting latent energy of the natural world. PMID:25394406

  6. Size and Carbon Content of Sub-seafloor Microbial Cells

    NASA Astrophysics Data System (ADS)

    Braun, S.; Morono, Y.; Littmann, S.; Jørgensen, B. B.; Lomstein, B. A.

    2015-12-01

    Into the seafloor, a radical decline in nutrient and energy availability poses strong metabolic demands to any residing life. However, a sedimentary microbial ecosystem seems to maintain itself close to what we understand to be the energetic limit of life. Since a complex sediment matrix is interfering with the analysis of whole cells and sub-cellular compounds such as cell wall and membrane molecules, little is known about the physiological properties of cells in the deep biosphere. Here we focus on the size and carbon content of cells from a 90-m sediment drill core retrieved in October 2013 at Landsort Deep, Baltic Sea, in 437 meters water depth. To determine their shape and volume, cells were separated from the sediment matrix by multi-layer density centrifugation and visualized via fluorescence microscopy (FM), scanning electron microscopy (SEM), and stimulated emission depletion microscopy (STED). Total cell-carbon was calculated from amino acid-carbon, which was analyzed by high-performance liquid chromatography after cells had additionally been purified by fluorescence activated cell sorting (FACS). Cell-carbon turnover times were estimated using an amino acid racemization model that is based on the built-in molecular clock of aspartic acid, which due to racemization alternates between the D- and L-isomeric configurations over timescales of thousands of years at low in-situ temperatures (≈4˚C). We find that the majority of microbial cells in the sediment have coccoid or rod-shaped morphology, and that absolute values for cell volume are strongly dependent on the method used, spanning three orders of magnitude from approximately 0.001 to 1 µm3 for both coccoid and rod-shaped cells. From the surface to the deepest sample measured (≈60 mbsf), cell volume decreases by an order of magnitude, and carbon content is in the lower range (<20 fg C cell-1) of what has been reported in the literature as conversion factors. Cell-carbon is turned over approximately

  7. High power density yeast catalyzed microbial fuel cells

    NASA Astrophysics Data System (ADS)

    Ganguli, Rahul

    Microbial fuel cells leverage whole cell biocatalysis to convert the energy stored in energy-rich renewable biomolecules such as sugar, directly to electrical energy at high efficiencies. Advantages of the process include ambient temperature operation, operation in natural streams such as wastewater without the need to clean electrodes, minimal balance-of-plant requirements compared to conventional fuel cells, and environmentally friendly operation. These make the technology very attractive as portable power sources and waste-to-energy converters. The principal problem facing the technology is the low power densities compared to other conventional portable power sources such as batteries and traditional fuel cells. In this work we examined the yeast catalyzed microbial fuel cell and developed methods to increase the power density from such fuel cells. A combination of cyclic voltammetry and optical absorption measurements were used to establish significant adsorption of electron mediators by the microbes. Mediator adsorption was demonstrated to be an important limitation in achieving high power densities in yeast-catalyzed microbial fuel cells. Specifically, the power densities are low for the length of time mediator adsorption continues to occur. Once the mediator adsorption stops, the power densities increase. Rotating disk chronoamperometry was used to extract reaction rate information, and a simple kinetic expression was developed for the current observed in the anodic half-cell. Since the rate expression showed that the current was directly related to microbe concentration close to the electrode, methods to increase cell mass attached to the anode was investigated. Electrically biased electrodes were demonstrated to develop biofilm-like layers of the Baker's yeast with a high concentration of cells directly connected to the electrode. The increased cell mass did increase the power density 2 times compared to a non biofilm fuel cell, but the power density

  8. Chemical modulation of the ultra-weak photon emission from Saccharomyces cerevisiae and differentiated HL-60 cells

    NASA Astrophysics Data System (ADS)

    Červinková, Kateřina; Nerudová, Michaela; Hašek, Jiří; Cifra, Michal

    2015-01-01

    The ultra-weak photon emission (UPE) is a universal phenomenon common to all cells with active oxidative metabolism. Generally accepted mechanism of the origin of the ultra-weak photon emission considers reactions of radical or nonradical reactive oxygen species (ROS) with biomolecules such as lipids and proteins which lead to the formation of electron excited species. During the transition to the ground state the excess energy is released as a photon with a wavelength in the visible range of the electromagnetic spectrum. Since the intensity of the light is very low it is possible to be measured only by highly sensitive devices. We used Hamamatsu Photonics PMT module H7360-01 mounted into a light-tight chamber for the purposes of this work. The goal of our research is to delineate an origin of UPE from two model organisms; differentiated HL-60 cells (human promyelocytic leukemia) and yeast cells Saccharomyces cerevisiae. While the UPE from the yeast cells arises spontaneously during the growth without any external stimuli, UPE from HL-60 is induced by phorbol 12-myristate, 13-acetate (PMA). It is possible to modulate the UPE production by certain antioxidants which scavenge ROS formed during the metabolism (yeast cells) or respiratory burst (HL-60 cells). The experiments are focused on the description of effects caused by antioxidants. Several kinds of antioxidants (ascorbic acid, mannitol, glutathione) with different concentration were used and we studied the changes in the UPE intensities of and the temporal developments of the optical signal.

  9. The Cytosolic pH of Individual Saccharomyces cerevisiae Cells Is a Key Factor in Acetic Acid Tolerance

    PubMed Central

    Fernández-Niño, Miguel; Marquina, Maribel; Swinnen, Steve; Rodríguez-Porrata, Boris

    2015-01-01

    It was shown recently that individual cells of an isogenic Saccharomyces cerevisiae population show variability in acetic acid tolerance, and this variability affects the quantitative manifestation of the trait at the population level. In the current study, we investigated whether cell-to-cell variability in acetic acid tolerance could be explained by the observed differences in the cytosolic pHs of individual cells immediately before exposure to the acid. Results obtained with cells of the strain CEN.PK113-7D in synthetic medium containing 96 mM acetic acid (pH 4.5) showed a direct correlation between the initial cytosolic pH and the cytosolic pH drop after exposure to the acid. Moreover, only cells with a low initial cytosolic pH, which experienced a less severe drop in cytosolic pH, were able to proliferate. A similar correlation between initial cytosolic pH and cytosolic pH drop was also observed in the more acid-tolerant strain MUCL 11987-9. Interestingly, a fraction of cells in the MUCL 11987-9 population showed initial cytosolic pH values below the minimal cytosolic pH detected in cells of the strain CEN.PK113-7D; consequently, these cells experienced less severe drops in cytosolic pH. Although this might explain in part the difference between the two strains with regard to the number of cells that resumed proliferation, it was observed that all cells from strain MUCL 11987-9 were able to proliferate, independently of their initial cytosolic pH. Therefore, other factors must also be involved in the greater ability of MUCL 11987-9 cells to endure strong drops in cytosolic pH. PMID:26341199

  10. Microbial solar cells: applying photosynthetic and electrochemically active organisms.

    PubMed

    Strik, David P B T B; Timmers, Ruud A; Helder, Marjolein; Steinbusch, Kirsten J J; Hamelers, Hubertus V M; Buisman, Cees J N

    2011-01-01

    Microbial solar cells (MSCs) are recently developed technologies that utilize solar energy to produce electricity or chemicals. MSCs use photoautotrophic microorganisms or higher plants to harvest solar energy, and use electrochemically active microorganisms in the bioelectrochemical system to generate electrical current. Here, we review the principles and performance of various MSCs in an effort to identify the most promising systems, as well as the bottlenecks and potential solutions, for "real-life" MSC applications. We present an outlook on future applications based on the intrinsic advantages of MSCs, specifically highlighting how these living energy systems can facilitate the development of an electricity-producing green roof. PMID:21067833

  11. An overview of electrode materials in microbial fuel cells

    NASA Astrophysics Data System (ADS)

    Zhou, Minghua; Chi, Meiling; Luo, Jianmei; He, Huanhuan; Jin, Tao

    2011-05-01

    Electrode materials play an important role in the performance (e.g., power output) and cost of microbial fuel cells (MFCs), which use bacteria as the catalysts to oxidize organic (inorganic) matter and convert chemical energy into electricity. In this paper, the recent progress of anode/cathode materials and filling materials as three-dimensional electrodes for MFCs has been systematically reviewed, resulting in comprehensive insights into the characteristics, options, modifications, and evaluations of the electrode materials and their effects on different actual wastewater treatment. Some existing problems of electrode materials in current MFCs are summarized, and outlooks for future development are also suggested.

  12. Microbial fuel cells as pollutant treatment units: Research updates.

    PubMed

    Zhang, Quanguo; Hu, Jianjun; Lee, Duu-Jong

    2016-10-01

    Microbial fuel cells (MFC) are a device that can convert chemical energy in influent substances to electricity via biological pathways. Based on the consent that MFC technology should be applied as a waste/wastewater treatment unit rather than a renewable energy source, this mini-review discussed recent R&D efforts on MFC technologies for pollutant treatments and highlighted the challenges and research and development needs. Owing to the low power density levels achievable by larger-scale MFC, the MFC should be used as a device other than energy source such as being a pollutant treatment unit. PMID:26906446

  13. Stacked microbial desalination cells to enhance water desalination efficiency.

    PubMed

    Chen, Xi; Xia, Xue; Liang, Peng; Cao, Xiaoxin; Sun, Haotian; Huang, Xia

    2011-03-15

    Microbial desalination cell (MDC) is a new method to obtain clean water from brackish water using electricity generated from organic matters by exoelectrogenic bacteria. Anions and cations, derived from salt solution filled in the desalination chamber between the anode and cathode, move to the anode and cathode chambers under the force of electrical field, respectively. On the basis of the primitive single-desalination-chambered MDC, stacked microbial desalination cells (SMDCs) were developed in order to promote the desalination rate in the present study. The effects of desalination chamber number and external resistance were investigated. Results showed that a remarkable increase in the total desalination rate (TDR) could be obtained by means of increasing the desalination cell number and reducing the external resistance, which caused the charge transfer efficiency increased since the SMDCs enabled more pairs of ions separated while one electron passed through the external circuit. The maximum TDR of 0.0252 g/h was obtained using a two-desalination-chambered SMDC with an external resistance of 10 Ω, which was 1.4 times that of single-desalination-chambered MDC. SMDCs proved to be an effective approach to increase the total water desalination rate if provided a proper desalination chamber number and external resistance. PMID:21322552

  14. Characterization of the Cell Surface Properties of Drinking Water Pathogens by Microbial Adhesion to Hydrocarbon and Electrophoretic Mobility Measurements

    EPA Science Inventory

    The surface characteristics of microbial cells directly influence their mobility and behavior within aqueous environments. The cell surface hydrophobicity (CSH) and electrophoretic mobility (EPM) of microbial cells impact a number of interactions and processes including aggregati...

  15. Cell-to-cell contact and antimicrobial peptides play a combined role in the death of Lachanchea thermotolerans during mixed-culture alcoholic fermentation with Saccharomyces cerevisiae.

    PubMed

    Kemsawasd, Varongsiri; Branco, Patrícia; Almeida, Maria Gabriela; Caldeira, Jorge; Albergaria, Helena; Arneborg, Nils

    2015-07-01

    The roles of cell-to-cell contact and antimicrobial peptides in the early death of Lachanchea thermotolerans CBS2803 during anaerobic, mixed-culture fermentations with Saccharomyces cerevisiae S101 were investigated using a commercially available, double-compartment fermentation system separated by cellulose membranes with different pore sizes, i.e. 1000 kDa for mixed- and single-culture fermentations, and 1000 and 3.5-5 kDa for compartmentalized-culture fermentations. SDS-PAGE and gel filtration chromatography were used to determine an antimicrobial peptidic fraction in the fermentations. Our results showed comparable amounts of the antimicrobial peptidic fraction in the inner compartments of the mixed-culture and 1000 kDa compartmentalized-culture fermentations containing L. thermotolerans after 4 days of fermentation, but a lower death rate of L. thermotolerans in the 1000 kDa compartmentalized-culture fermentation than in the mixed-culture fermentation. Furthermore, L. thermotolerans died off even more slowly in the 3.5-5 kDa than in the 1000 kDa compartmentalized-culture fermentation, which coincided with the presence of less of the antimicrobial peptidic fraction in the inner compartment of that fermentation than of the 1000 kDa compartmentalized-culture fermentation. Taken together, these results indicate that the death of L. thermotolerans in mixed cultures with S. cerevisiae is caused by a combination of cell-to-cell contact and antimicrobial peptides. PMID:26109361

  16. The Role of Soil Organic Matter, Nutrients, and Microbial Community Structure on the Performance of Microbial Fuel Cells

    NASA Astrophysics Data System (ADS)

    Rooney-Varga, J. N.; Dunaj, S. J.; Vallino, J. J.; Hines, M. E.; Gay, M.; Kobyljanec, C.

    2011-12-01

    Microbial fuel cells (MFCs) offer the potential for generating electricity, mitigating greenhouse gas emissions, and bioremediating pollutants through utilization of a plentiful, natural, and renewable resource: soil organic carbon. In the current study, we analyzed microbial community structure, MFC performance, and soil characteristics in different microhabitats (bulk soil, anode, and cathode) within MFCs constructed from agricultural or forest soils in order to determine how soil type and microbial dynamics influence MFC performance. MFCs were constructed with soils from agricultural and hardwood forest sites at Harvard Forest (Petersham, MA). The bulk soil characteristics were analyzed, including polyphenols, short chain fatty acids, total organic C and N, abiotic macronutrients, N and P mineralization rates, CO2 respiration rates, and MFC power output. Microbial community structure of the anodes, cathodes, and bulk soils was determined with molecular fingerprinting methods, which included terminal restriction length polymorphism (T-RFLP) analysis and 16S rRNA gene sequencing analysis. Our results indicated that MFCs constructed from agricultural soil had power output about 17 times that of forest soil-based MFCs and respiration rates about 10 times higher than forest soil MFCs. Agricultural soil MFCs had lower C:N ratios, polyphenol content, and acetate concentrations than forest soil MFCs, suggesting that active agricultural MFC microbial communities were supported by higher quality organic carbon. Microbial community profile data indicate that the microbial communities at the anode of the high power MFCs were less diverse than in low power MFCs and were dominated by Deltaproteobacteria, Geobacter, and, to a lesser extent, Clostridia, while low-power MFC anode communities were dominated by Clostridia. These data suggest that the presence of organic carbon substrate (acetate) was not the major limiting factor in selecting for highly electrogenic microbial

  17. Monitoring microbial population dynamics at low densities

    NASA Astrophysics Data System (ADS)

    Julou, Thomas; Desprat, Nicolas; Bensimon, David; Croquette, Vincent

    2012-07-01

    We propose a new and simple method for the measurement of microbial concentrations in highly diluted cultures. This method is based on an analysis of the intensity fluctuations of light scattered by microbial cells under laser illumination. Two possible measurement strategies are identified and compared using simulations and measurements of the concentration of gold nanoparticles. Based on this comparison, we show that the concentration of Escherichia coli and Saccharomyces cerevisiae cultures can be easily measured in situ across a concentration range that spans five orders of magnitude. The lowest measurable concentration is three orders of magnitude (1000×) smaller than in current optical density measurements. We show further that this method can also be used to measure the concentration of fluorescent microbial cells. In practice, this new method is well suited to monitor the dynamics of population growth at early colonization of a liquid culture medium. The dynamic data thus obtained are particularly relevant for microbial ecology studies.

  18. Microbial reverse electrodialysis cells for synergistically enhanced power production.

    PubMed

    Kim, Younggy; Logan, Bruce E

    2011-07-01

    A new type of bioelectrochemical system for producing electrical power, called a microbial reverse-electrodialysis cell (MRC), was developed to increase voltages and power densities compared to those generated individually by microbial fuel cells (MFCs) or reverse electrodialysis (RED) systems. In RED systems, electrode overpotentials create significant energy losses due to thermodynamically unfavorable electrode reactions, and therefore a large number of stacked cells must be used to have significant energy recovery. This results in high capital costs for the large number of membranes, and increases energy losses from pumping water through a large number of cells. In an MRC, high overpotentials are avoided through oxidation of organic matter by exoelectrogenic bacteria on the anode and oxygen reduction on the cathode. An MRC containing only five pairs of RED cells, fed solutions typical of seawater (600 mM NaCl) and river water (12 mM NaCl) at 0.85 mL/min, produced up to 3.6 W/m(2) (cathode surface area) and 1.2-1.3 V with acetate as a substrate. Pumping accounted for <2% of the produced power. A higher flow rate (1.55 mL/min) increased power densities up to 4.3 W/m(2). COD removal was 98% with a Coulombic efficiency of 64%. Power production by the individual components was substantially lower with 0.7 W/m(2) without salinity driven energy, and <0.015 W/m(2) with reduced exoelectrogenic activity due to substrate depletion. These results show that the combination of an MFC and a RED stack synergistically increases performance relative to the individual systems, producing a new type of system that can be used to more efficiently capture salinity driven energy from seawater and river water. PMID:21644573

  19. DNA Damage Response Checkpoint Activation Drives KP1019 Dependent Pre-Anaphase Cell Cycle Delay in S. cerevisiae

    PubMed Central

    Bierle, Lindsey A.; Reich, Kira L.; Taylor, Braden E.; Blatt, Eliot B.; Middleton, Sydney M.; Burke, Shawnecca D.; Stultz, Laura K.; Hanson, Pamela K.; Partridge, Janet F.; Miller, Mary E.

    2015-01-01

    Careful regulation of the cell cycle is required for proper replication, cell division, and DNA repair. DNA damage–including that induced by many anticancer drugs–results in cell cycle delay or arrest, which can allow time for repair of DNA lesions. Although its molecular mechanism of action remains a matter of debate, the anticancer ruthenium complex KP1019 has been shown to bind DNA in biophysical assays and to damage DNA of colorectal and ovarian cancer cells in vitro. KP1019 has also been shown to induce mutations and induce cell cycle arrest in Saccharomyces cerevisiae, suggesting that budding yeast can serve as an appropriate model for characterizing the cellular response to the drug. Here we use a transcriptomic approach to verify that KP1019 induces the DNA damage response (DDR) and find that KP1019 dependent expression of HUG1 requires the Dun1 checkpoint; both consistent with KP1019 DDR in budding yeast. We observe a robust KP1019 dependent delay in cell cycle progression as measured by increase in large budded cells, 2C DNA content, and accumulation of Pds1 which functions to inhibit anaphase. Importantly, we also find that deletion of RAD9, a gene required for the DDR, blocks drug-dependent changes in cell cycle progression, thereby establishing a causal link between the DDR and phenotypes induced by KP1019. Interestingly, yeast treated with KP1019 not only delay in G2/M, but also exhibit abnormal nuclear position, wherein the nucleus spans the bud neck. This morphology correlates with short, misaligned spindles and is dependent on the dynein heavy chain gene DYN1. We find that KP1019 creates an environment where cells respond to DNA damage through nuclear (transcriptional changes) and cytoplasmic (motor protein activity) events. PMID:26375390

  20. Alkali extraction of beta-d-glucans from Saccharomyces cerevisiae cell wall and study of their adsorptive properties toward zearalenone.

    PubMed

    Yiannikouris, Alexandros; François, Jean; Poughon, Laurent; Dussap, Claude-Gilles; Bertin, Gérard; Jeminet, Georges; Jouany, Jean-Pierre

    2004-06-01

    The isolated cell wall of Saccharomyces cerevisiae has some capacity to adsorb zearalenone (affinity near 30%) and reduce the bioavailability of toxins in the digestive tract. The adsorption process was quantified in vitro, and the data obtained when plotted with Hill's equation indicated a cooperative process. The model showed that the adsorption capacity was related to the yeast cell wall composition. This work focused on the role of various beta-d-glucan types in the efficacy of zearalenone adsorption by yeast cell wall and sought to elucidate some of the adsorption mechanisms. Zearalenone was mixed at 37 degrees C with a constant quantity of alkali-soluble or alkali-insoluble beta-d-glucans isolated from yeast cell walls, and the amount of adsorbed zearalenone was measured. Given that the alkali solubility of beta-d-glucans is a determining factor for their three-dimensional conformation and that the alkali-insoluble fraction had a greater affinity (up to 50%) than the alkali-soluble fraction ( approximately 16%), it was concluded that the three-dimensional structure strongly influences the adsorption process. The alkali insolubility of beta-d-glucans led to the formation of single and/or triple helices, which have been identified as the most favorable structures for zearalenone adsorption efficacy. The beta(1,3)-d-glucan and beta(1,6)-d-glucan compositions of the two alkali-extracted fractions and their involvement in the adsorption process are discussed. PMID:15161247

  1. First steps towards a constructal Microbial Fuel Cell.

    PubMed

    Lepage, Guillaume; Perrier, Gérard; Ramousse, Julien; Merlin, Gérard

    2014-06-01

    In order to reach real operating conditions with consequent organic charge flow, a multi-channel reactor for Microbial Fuel Cells is designed. The feed-through double chamber reactor is a two-dimensional system with four parallel channels and Reticulated Vitreous Carbon as electrodes. Based on thermodynamical calculations, the constructal-inspired distributor is optimized with the aim to reduce entropy generation along the distributing path. In the case of negligible singular pressure drops, the Hess-Murray law links the lengths and the hydraulic diameters of the successive reducing ducts leading to one given working channel. The determination of generated entropy in the channels of our constructal MFC is based on the global hydraulic resistance caused by both regular and singular pressure drops. Polarization, power and Electrochemical Impedance Spectroscopy show the robustness and the efficiency of the cell, and therefore the potential of the constructal approach. Routes towards improvements are suggested in terms of design evolutions. PMID:24747390

  2. Low external pH induces HOG1-dependent changes in the organization of the Saccharomyces cerevisiae cell wall.

    PubMed

    Kapteyn, J C; ter Riet, B; Vink, E; Blad, S; De Nobel, H; Van Den Ende, H; Klis, F M

    2001-01-01

    Low environmental pH strongly affected the organization of the Saccharomyces cerevisiae cell wall, resulting in rapidly induced resistance to beta1,3-glucanase. At a molecular level, we found that a considerable amount of Cwp1p became anchored through a novel type of linkage for glycosylphosphatidylinositol (GPI)-dependent cell wall proteins, namely an alkali-labile linkage to beta1,3-glucan. This novel type of modification for Cwp1p did not require the presence of a GPI-derived structure connecting the protein with beta1,6-glucan. In addition, we found high levels of Cwp1p, which was double-anchored through both the novel alkali-sensitive bond to beta1,3-glucan and the alkali-resistant GPI-derived linkage to beta1,6-glucan. Further cell wall analyses demonstrated that Pir2p/Hsp150 and possibly other Pir cell wall proteins, which were already known to be linked to the beta1,3-glucan framework by an alkali-sensitive linkage, were also more efficiently retained in the cell wall at pH 3.5 than at pH 5.5. Consequently, the alkali-sensitive type of linkage of cell wall proteins to beta1,3-glucan was induced by low pH. The low pH-induced alterations in yeast cell wall architecture were demonstrated to be dependent on a functional HOG1 gene, but not on the Slt2p-mediated MAP kinase pathway. Consistent with this observation, DNA microarray studies revealed transcriptional induction of many known high-osmolarity glycerol (HOG) pathway-dependent genes, including four cell wall-related genes, namely CWP1, HOR7, SPI1 and YGP1. PMID:11136466

  3. Enhancement of Electricity Production by Graphene Oxide in Soil Microbial Fuel Cells and Plant Microbial Fuel Cells

    PubMed Central

    Goto, Yuko; Yoshida, Naoko; Umeyama, Yuto; Yamada, Takeshi; Tero, Ryugo; Hiraishi, Akira

    2015-01-01

    The effects of graphene oxide (GO) on electricity generation in soil microbial fuel cells (SMFCs) and plant microbial fuel cell (PMFCs) were investigated. GO at concentrations ranging from 0 to 1.9 g⋅kg−1 was added to soil and reduced for 10 days under anaerobic incubation. All SMFCs (GO-SMFCs) utilizing the soils incubated with GO produced electricity at a greater rate and in higher quantities than the SMFCs which did not contain GO. In fed-batch operations, the overall average electricity generation in GO-SMFCs containing 1.0 g⋅kg−1 of GO was 40 ± 19 mW⋅m−2, which was significantly higher than the value of 6.6 ± 8.9 mW⋅m−2 generated from GO-free SMFCs (p < 0.05). The increase in catalytic current at the oxidative potential was observed by cyclic voltammetry (CV) for GO-SMFC, with the CV curve suggesting the enhancement of electron transfer from oxidation of organic substances in the soil by the reduced form of GO. The GO-containing PMFC also displayed a greater generation of electricity compared to the PMFC with no added GO, with GO-PMFC producing 49 mW⋅m−2 of electricity after 27 days of operation. Collectively, this study demonstrates that GO added to soil can be microbially reduced in soil, and facilitates electron transfer to the anode in both SMFCs and PMFCs. PMID:25883931

  4. [Production of β-carotene by metabolically engineered Saccharomyces cerevisiae].

    PubMed

    Wang, Beibei; Shi, Mingyu; Wang, Dong; Xu, Jiaoyang; Liu, Yi; Yang, Hongjiang; Dai, Zhubo; Zhang, Xueli

    2014-08-01

    β-carotene has a wide range of application in food, pharmaceutical and cosmetic industries. For microbial production of β-carotene in Saccharomyces cerevisiae, the supply of geranylgeranyl diphosphate (GGPP) was firstly increased in S. cerevisiae BY4742 to obtain strain BY4742-T2 through over-expressing truncated 3-hydroxy-3-methylglutaryl-CoA reductase (tHMGR), which is the major rate-limiting enzyme in the mevalonate (MVA) pathway, and GGPP synthase (GGPS), which is a key enzyme in the diterpenoid synthetic pathway. The β-carotene synthetic genes of Pantoea agglomerans and Xanthophyllomyces dendrorhous were further integrated into strain BY4742-T2 for comparing β-carotene production. Over-expression of tHMGR and GGPS genes led to 26.0-fold increase of β-carotene production. In addition, genes from X. dendrorhous was more efficient than those from P. agglomerans for β-carotene production in S. cerevisiae. Strain BW02 was obtained which produced 1.56 mg/g (dry cell weight) β-carotene, which could be used further for constructing cell factories for β-carotene production. PMID:25507473

  5. [Production of β-carotene by metabolically engineered Saccharomyces cerevisiae].

    PubMed

    Wang, Beibei; Shi, Mingyu; Wang, Dong; Xu, Jiaoyang; Liu, Yi; Yang, Hongjiang; Dai, Zhubo; Zhang, Xueli

    2014-08-01

    β-carotene has a wide range of application in food, pharmaceutical and cosmetic industries. For microbial production of β-carotene in Saccharomyces cerevisiae, the supply of geranylgeranyl diphosphate (GGPP) was firstly increased in S. cerevisiae BY4742 to obtain strain BY4742-T2 through over-expressing truncated 3-hydroxy-3-methylglutaryl-CoA reductase (tHMGR), which is the major rate-limiting enzyme in the mevalonate (MVA) pathway, and GGPP synthase (GGPS), which is a key enzyme in the diterpenoid synthetic pathway. The β-carotene synthetic genes of Pantoea agglomerans and Xanthophyllomyces dendrorhous were further integrated into strain BY4742-T2 for comparing β-carotene production. Over-expression of tHMGR and GGPS genes led to 26.0-fold increase of β-carotene production. In addition, genes from X. dendrorhous was more efficient than those from P. agglomerans for β-carotene production in S. cerevisiae. Strain BW02 was obtained which produced 1.56 mg/g (dry cell weight) β-carotene, which could be used further for constructing cell factories for β-carotene production. PMID:25423750

  6. Direct observation of oxidative stress on the cell wall of Saccharomyces cerevisiae strains with atomic force microscopy.

    PubMed

    de Souza Pereira, R; Geibel, J

    1999-11-01

    We imaged pores on the surface of the cell wall of three different industrial strains of Saccharomyces cerevisiae using atomic force microscopy. The pores could be enlarged using 10 mM diamide, an SH residue oxidant that attacks surface proteins. We found that two strains showed signs of oxidative damage via changes in density and diameter of the surface pores. We found that the German strain was resistant to diamide induced oxidative damage, even when the concentration of the oxidant was increased to 50 mM. The normal pore size found on the cell walls of American strains had diameters of about 200 nm. Under conditions of oxidative stress the diameters changed to 400 nm. This method may prove to be a useful rapid screening process (45-60 min) to determine which strains are oxidative resistant, as well as being able to screen for groups of yeast that are sensitive to oxidative stress. This rapid screening tool may have direct applications in molecular biology (transference of the genes to inside of living cells) and biotechnology (biotransformations reactions to produce chiral synthons in organic chemistry. PMID:10630618

  7. RAD9-dependent G1 arrest defines a second checkpoint for damaged DNA in the cell cycle of Saccharomyces cerevisiae.

    PubMed Central

    Siede, W; Friedberg, A S; Friedberg, E C

    1993-01-01

    Exposure of the yeast Saccharomyces cerevisiae to ultraviolet (UV) light, the UV-mimetic chemical 4-nitroquinoline-1-oxide (4NQO), or gamma radiation after release from G1 arrest induced by alpha factor results in delayed resumption of the cell cycle. As is the case with G2 arrest following ionizing radiation damage [Weinert, T. A. & Hartwell, L. H. (1988) Science 241, 317-322], the normal execution of DNA damage-induced G1 arrest depends on a functional yeast RAD9 gene. We suggest that the RAD9 gene product may interact with cellular components common to the G1/S and G2/M transition points in the cell cycle of this yeast. These observations define a checkpoint in the eukaryotic cell cycle that may facilitate the repair of lesions that are otherwise processed to lethal and/or mutagenic damage during DNA replication. This checkpoint apparently operates after the mating pheromone-induced G1 arrest point but prior to replicative DNA synthesis, S phase-associated maximal induction of histone H2A mRNA, and bud emergence. Images Fig. 4 PMID:8367452

  8. The Saccharomyces cerevisiae RNase mitochondrial RNA processing is critical for cell cycle progression at the end of mitosis.

    PubMed Central

    Cai, Ti; Aulds, Jason; Gill, Tina; Cerio, Michael; Schmitt, Mark E

    2002-01-01

    We have identified a cell cycle delay in Saccharomyces cerevisiae RNase MRP mutants. Mutants delay with large budded cells, dumbbell-shaped nuclei, and extended spindles characteristic of "exit from mitosis" mutants. In accord with this, a RNase MRP mutation can be suppressed by overexpressing the polo-like kinase CDC5 or by deleting the B-type cyclin CLB1, without restoring the MRP-dependent rRNA-processing step. In addition, we identified a series of genetic interactions between RNase MRP mutations and mutations in CDC5, CDC14, CDC15, CLB2, and CLB5. As in most "exit from mitosis" mutants, levels of the Clb2 cyclin were increased. The buildup of Clb2 protein is not the result of a defect in the release of the Cdc14 phosphatase from the nucleolus, but rather the result of an increase in CLB2 mRNA levels. These results indicate a clear role of RNase MRP in cell cycle progression at the end of mitosis. Conservation of this function in humans may explain many of the pleiotropic phenotypes of cartilage hair hypoplasia. PMID:12136008

  9. Towards an engineering-oriented strategy for building microbial anodes for microbial fuel cells.

    PubMed

    Pocaznoi, Diana; Erable, Benjamin; Etcheverry, Luc; Delia, Marie-Line; Bergel, Alain

    2012-10-14

    The objective of the work was to give some first insight into an engineering-oriented approach to MFC design by focusing on anode optimisation. The effect of various parameters was firstly investigated in half cell set-ups under well-controlled conditions. Microbial anodes were formed from soil leachate under polarisation at -0.2 V vs. SCE with different concentrations of substrate, salt and buffer. It was shown that non-turnover CV could be used to assess the electroactive maturity of the anodes during polarisation. This first phase resulted in the definition of a set of optimal parameter values. In the second phase, an optimal anode was formed in a half-cell under the defined optimal conditions. A numerical approach was then developed to calculate the theoretical maximum power that the anode could provide in an ideal MFC. The concept of "ideal MFC" introduced here allowed the theoretical maximum power to be calculated on the sole basis of the kinetic characteristics of the anode. Finally, a MFC designed in the aim of approaching such ideal conditions generated stable power densities of 6.0 W m(-2), which were among the highest values reported so far. The discrepancy between the theoretical maximum (8.9 W m(-2)) and the experimental results pointed out some limit due to the source of inoculum and suggested possible paths to improvement. PMID:22932946

  10. Enhancement of gene targeting in human cells by intranuclear permeation of the Saccharomyces cerevisiae Rad52 protein

    PubMed Central

    Kalvala, Arjun; Rainaldi, Giuseppe; Di Primio, Cristina; Liverani, Vania; Falaschi, Arturo; Galli, Alvaro

    2010-01-01

    The introduction of exogenous DNA in human somatic cells results in a frequency of random integration at least 100-fold higher than gene targeting (GT), posing a seemingly insurmountable limitation for gene therapy applications. We previously reported that, in human cells, the stable over-expression of the Saccharomyces cerevisiae Rad52 gene (yRAD52), which plays the major role in yeast homologous recombination (HR), caused an up to 37-fold increase in the frequency of GT, indicating that yRAD52 interacts with the double-strand break repair pathway(s) of human cells favoring homologous integration. In the present study, we tested the effect of the yRad52 protein by delivering it directly to the human cells. To this purpose, we fused the yRAD52 cDNA to the arginine-rich domain of the TAT protein of HIV (tat11) that is known to permeate the cell membranes. We observed that a recombinant yRad52tat11 fusion protein produced in Escherichia coli, which maintains its ability to bind single-stranded DNA (ssDNA), enters the cells and the nuclei, where it is able to increase both intrachromosomal recombination and GT up to 63- and 50-fold, respectively. Moreover, the non-homologous plasmid DNA integration decreased by 4-fold. yRAD52tat11 proteins carrying point mutations in the ssDNA binding domain caused a lower or nil increase in recombination proficiency. Thus, the yRad52tat11 could be instrumental to increase GT in human cells and a ‘protein delivery approach’ offers a new tool for developing novel strategies for genome modification and gene therapy applications. PMID:20519199

  11. Atomic Force Microscopy in Microbiology: New Structural and Functional Insights into the Microbial Cell Surface

    PubMed Central

    2014-01-01

    ABSTRACT Microbial cells sense and respond to their environment using their surface constituents. Therefore, understanding the assembly and biophysical properties of cell surface molecules is an important research topic. With its ability to observe living microbial cells at nanometer resolution and to manipulate single-cell surface molecules, atomic force microscopy (AFM) has emerged as a powerful tool in microbiology. Here, we survey major breakthroughs made in cell surface microbiology using AFM techniques, emphasizing the most recent structural and functional insights. PMID:25053785

  12. Complete cobalt recovery from lithium cobalt oxide in self-driven microbial fuel cell - Microbial electrolysis cell systems

    NASA Astrophysics Data System (ADS)

    Huang, Liping; Yao, Binglin; Wu, Dan; Quan, Xie

    2014-08-01

    Complete cobalt recovery from lithium cobalt oxide requires to firstly leach cobalt from particles LiCoO2 and then recover cobalt from aqueous Co(II). A self-driven microbial fuel cell (MFC)-microbial electrolysis cell (MEC) system can completely carry out these two processes, in which Co(II) is firstly released from particles LiCoO2 on the cathodes of MFCs and then reduced on the cathodes of MECs which are powered by the cobalt leaching MFCs. A cobalt leaching rate of 46 ± 2 mg L-1 h-1 with yield of 1.5 ± 0.1 g Co g-1 COD (MFCs) and a Co(II) reduction rate of 7 ± 0 mg L-1 h-1 with yield of 0.8 ± 0.0 g Co g-1 COD (MECs), as well as a overall system cobalt yield of 0.15 ± 0.01 g Co g-1 Co can be achieved in this self-driven MFC-MEC system. Coulombic efficiencies reach 41 ± 1% (anodic MFCs), 75 ± 0% (anodic MECs), 100 ± 2% (cathodic MFCs), and 29 ± 1% (cathodic MECs) whereas overall system efficiency averages 34 ± 1%. These results provide a new process of linking MFCs to MECs for complete recovery of cobalt and recycle of spent lithium ion batteries with no external energy consumption.

  13. A comparison of simultaneous organic carbon and nitrogen removal in microbial fuel cells and microbial electrolysis cells.

    PubMed

    Hussain, Abid; Manuel, Michelle; Tartakovsky, Boris

    2016-05-15

    This study demonstrates simultaneous carbon and nitrogen removal in laboratory-scale continuous flow microbial fuel cell (MFC) and microbial electrolysis cell (MEC) and provides side-by side comparison of these bioelectrochemical systems. The maximum organic carbon removal rates in MFC and MEC tests were similar at 5.1 g L(-1) d(-1) and 4.16 g L(-1) d(-1), respectively, with a near 100% carbon removal efficiency at an organic load of 3.3 g L(-1) d(-1). An ammonium removal efficiency of 30-55% with near-zero nitrite and nitrate concentrations was observed in the MFC operated at an optimal external resistance, while open-circuit MFC operation resulted in a reduced carbon and ammonium removal of 53% and 21%, respectively. In the MEC ammonium removal was limited to 7-12% under anaerobic conditions, while micro-aerobic conditions increased the removal efficiency to 31%. Also, at zero applied voltage both carbon and ammonium removal efficiencies were reduced to 42% and 4%, respectively. Based on the observed performance under different operating conditions, it was concluded that simultaneous carbon and nitrogen removal was facilitated by concurrent anaerobic and aerobic biotransformation pathways at the anode and cathode, which balanced bioelectrochemical nitrification and denitrification reactions. PMID:26950500

  14. Integrated hydrogen production process from cellulose by combining dark fermentation, microbial fuel cells, and a microbial electrolysis cell.

    PubMed

    Wang, Aijie; Sun, Dan; Cao, Guangli; Wang, Haoyu; Ren, Nanqi; Wu, Wei-Min; Logan, Bruce E

    2011-03-01

    Hydrogen gas production from cellulose was investigated using an integrated hydrogen production process consisting of a dark fermentation reactor and microbial fuel cells (MFCs) as power sources for a microbial electrolysis cell (MEC). Two MFCs (each 25 mL) connected in series to an MEC (72 mL) produced a maximum of 0.43 V using fermentation effluent as a feed, achieving a hydrogen production rate from the MEC of 0.48 m(3) H(2)/m(3)/d (based on the MEC volume), and a yield of 33.2 mmol H(2)/g COD removed in the MEC. The overall hydrogen production for the integrated system (fermentation, MFC and MEC) was increased by 41% compared with fermentation alone to 14.3 mmol H(2)/g cellulose, with a total hydrogen production rate of 0.24 m(3) H(2)/m(3)/d and an overall energy recovery efficiency of 23% (based on cellulose removed) without the need for any external electrical energy input. PMID:21216594

  15. Self-sustained reduction of multiple metals in a microbial fuel cell-microbial electrolysis cell hybrid system.

    PubMed

    Li, Yan; Wu, Yining; Liu, Bingchuan; Luan, Hongwei; Vadas, Timothy; Guo, Wanqian; Ding, Jie; Li, Baikun

    2015-09-01

    A self-sustained hybrid bioelectrochemical system consisting of microbial fuel cell (MFC) and microbial electrolysis cell (MEC) was developed to reduce multiple metals simultaneously by utilizing different reaction potentials. Three heavy metals representing spontaneous reaction (chromium, Cr) and unspontaneous reaction (lead, Pb and nickel, Ni) were selected in this batch-mode study. The maximum power density of the MFC achieved 189.4 mW m(-2), and the energy recovery relative to the energy storage circuit (ESC) was ∼ 450%. At the initial concentration of 100 mg L(-1), the average reduction rate of Cr(VI) was 30.0 mg L(-1) d(-1), Pb(II) 32.7 mg L(-1) d(-1), and Ni(II) 8.9 mg L(-1) d(-1). An electrochemical model was developed to predict the change of metal concentration over time. The power output of the MFC was sufficient to meet the requirement of the ESC and MEC, and the "self-sustained metal reduction" was achieved in this hybrid system. PMID:26038328

  16. Molecular Basis for Strain Variation in the Saccharomyces cerevisiae Adhesin Flo11p.

    PubMed

    Barua, Subit; Li, Li; Lipke, Peter N; Dranginis, Anne M

    2016-01-01

    FLO11 encodes a yeast cell wall flocculin that mediates a variety of adhesive phenotypes in Saccharomyces cerevisiae. Flo11p is implicated in many developmental processes, including flocculation, formation of pseudohyphae, agar invasion, and formation of microbial mats and biofilms. However, Flo11p mediates different processes in different yeast strains. To investigate the mechanisms by which FLO11 determines these differences in colony morphology, flocculation, and invasion, we studied gene structure, function, and expression levels. Nonflocculent Saccharomyces cerevisiae Σ1278b cells exhibited significantly higher FLO11 mRNA expression, especially in the stationary phase, than highly flocculent S. cerevisiae var. diastaticus. The two strains varied in cell surface hydrophobicity, and Flo11p contributed significantly to surface hydrophobicity in S. cerevisiae var. diastaticus but not in strain Σ1278b. Sequencing of the FLO11 gene in S. cerevisiae var. diastaticus revealed strain-specific differences, including a 15-amino-acid insertion in the adhesion domain. Flo11p adhesion domains from strain Σ1278b and S. cerevisiae var. diastaticus were expressed and used to coat magnetic beads. The adhesion domain from each strain bound preferentially to homologous cells, and the preferences were independent of the cells in which the adhesion domains were produced. These results are consistent with the idea that strain-specific variations in the amino acid sequences in the adhesion domains cause different Flo11p flocculation activities. The results also imply that strain-specific differences in expression levels, posttranslational modifications, and allelic differences outside the adhesion domains have little effect on flocculation. IMPORTANCE As a nonmotile organism, Saccharomyces cerevisiae employs the cell surface flocculin Flo11/Muc1 as an important means of adapting to environmental change. However, there is a great deal of strain variation in the expression of

  17. Molecular Basis for Strain Variation in the Saccharomyces cerevisiae Adhesin Flo11p

    PubMed Central

    Li, Li; Lipke, Peter N.; Dranginis, Anne M.

    2016-01-01

    ABSTRACT FLO11 encodes a yeast cell wall flocculin that mediates a variety of adhesive phenotypes in Saccharomyces cerevisiae. Flo11p is implicated in many developmental processes, including flocculation, formation of pseudohyphae, agar invasion, and formation of microbial mats and biofilms. However, Flo11p mediates different processes in different yeast strains. To investigate the mechanisms by which FLO11 determines these differences in colony morphology, flocculation, and invasion, we studied gene structure, function, and expression levels. Nonflocculent Saccharomyces cerevisiae Σ1278b cells exhibited significantly higher FLO11 mRNA expression, especially in the stationary phase, than highly flocculent S. cerevisiae var. diastaticus. The two strains varied in cell surface hydrophobicity, and Flo11p contributed significantly to surface hydrophobicity in S. cerevisiae var. diastaticus but not in strain Σ1278b. Sequencing of the FLO11 gene in S. cerevisiae var. diastaticus revealed strain-specific differences, including a 15-amino-acid insertion in the adhesion domain. Flo11p adhesion domains from strain Σ1278b and S. cerevisiae var. diastaticus were expressed and used to coat magnetic beads. The adhesion domain from each strain bound preferentially to homologous cells, and the preferences were independent of the cells in which the adhesion domains were produced. These results are consistent with the idea that strain-specific variations in the amino acid sequences in the adhesion domains cause different Flo11p flocculation activities. The results also imply that strain-specific differences in expression levels, posttranslational modifications, and allelic differences outside the adhesion domains have little effect on flocculation. IMPORTANCE As a nonmotile organism, Saccharomyces cerevisiae employs the cell surface flocculin Flo11/Muc1 as an important means of adapting to environmental change. However, there is a great deal of strain variation in the

  18. Sorption of trivalent cerium by a mixture of microbial cells and manganese oxides: Effect of microbial cells on the oxidation of trivalent cerium

    NASA Astrophysics Data System (ADS)

    Ohnuki, Toshihiko; Jiang, Mingyu; Sakamoto, Fuminori; Kozai, Naofumi; Yamasaki, Shinya; Yu, Qianqian; Tanaka, Kazuya; Utsunomiya, Satoshi; Xia, Xiaobin; Yang, Ke; He, Jianhua

    2015-08-01

    Sorption of Ce by mixtures of synthetic Mn oxides and microbial cells of Pseudomonas fluorescens was investigated to elucidate the role of microorganisms on Ce(III) oxidative migration in the environment. The mixtures, upon which Ce was sorbed following exposure to solutions containing 1.0 × 10-4 or 1.0 × 10-5 mol L-1 Ce(III), were analyzed by scanning electron microscopy energy dispersive X-ray spectroscopy (SEM-EDS) and micro-X-ray fluorescence (micro-XRF) at synchrotron facilities. A Ce LIII-edge micro XANES spectra analysis was also performed to determine the oxidation states of Ce adsorbed to the Mn oxides and microbial cells in the mixtures. The distribution ratios (Kd) of Ce between the individual solids and solution increased with increasing pH of the solution, and was nearly the same in mixtures containing varying amounts of microbial cells. SEM-EDS and micro-XRF analyses showed that Ce was sorbed by both MnO2 and microbial cells (1.7 × 10-1 or 3.3 × 10-1 g L-1). In addition, nano-particles containing Ce and P developed on the surface of the microbial cells. XANES analysis showed that lower fractions of Ce(III) were oxidized to Ce(IV) in the mixtures containing greater amounts of microbial cells. Micro-XANES analysis revealed that Ce was present as Ce(III) on the microbial cells and as Ce(IV) on Mn oxides. These results strongly suggest that the association of Ce(III) with the microbial cell surface and the formation of Ce phosphate nano-particles are responsible for suppressing the oxidation of Ce(III) to Ce(IV) in the mixtures.

  19. Modification of a French pressure cell to improve microbial cell disruption.

    PubMed

    Jaschke, Paul R; Drake, Ian; Beatty, J Thomas

    2009-10-01

    A procedure for modification of the valve stem of a 40 K French pressure cell is described. The modification should be done by a machinist and requires a metalworking lathe. After modification of the valve stem, a torlon 4203 plastic ball is used between the valve stem and valve seat to control the pressure within the cell. The torlon plastic ball is a key component needed to obtain the high pressures required for efficient disruption of microbial cells. PMID:19731071

  20. Microbial Cell Budget of a High-Arctic Supraglacial Catchment

    NASA Astrophysics Data System (ADS)

    Irvine-Fynn, T. D.; Edwards, A.; Newton, S.; Langford, H.; Rassner, S. M.; Telling, J.; Anesio, A. M.; Hodson, A. J.

    2012-12-01

    There is a growing recognition of glaciers as ecosystems and a source of organic matter delivered to downstream environments. Recently, researchers have focussed on examination of interred cells entombed within the glacier body and the dissolved organic matter, particularly carbon, conveyed in meltwaters. However, due to a reliance on cell concentration measurements derived from ice cores rather than meltwater runoff, uncertainty surrounds the estimates of contributions in the form of microbial cells' particulate carbon liberated from glaciers. Here, using flow cytometry, we present the first enumeration of biological particles draining from a supraglacial catchment on Midtre Lovénbreen (Svalbard) over a 36-day study period. An average in-stream cell flux of 1.08×107 cells m-2 hr-1 was found. Non-linear associations between water discharge and biological particle concentrations were identified, which provides insight into glacier surface hydraulics. Crucially, contrast between ice-melt and aeolian inputs to, and the fluvial output from the monitored catchment suggested storage of 8.83×107 cells m-2 hr-1. The physical retention of particulates at glacier surfaces may contribute to mass thinning through the feedbacks altering surface ice albedo. Nonetheless, over the period of observation, 7.5×1014 cells were conveyed from the glacier, and allometric relationships between cells and nutrients allowed estimates of the corresponding carbon, protein and DNA delivery to downstream environments. This study demonstrates that interactions between biological processes and ice surface hydraulics merit further investigation not only for nutrient release, but also for better comprehension of mechanisms behind global ice mass wastage and the primary colonisation of newly exposed glacier forefields.raph illustrating discharge (Q) vs. supraglacial in-stream cell flux

  1. Genomic Sequencing of Single Microbial Cells from Environmental Samples

    SciTech Connect

    Ishoey, Thomas; Woyke, Tanja; Stepanauskas, Ramunas; Novotny, Mark; Lasken, Roger S.

    2008-02-01

    Recently developed techniques allow genomic DNA sequencing from single microbial cells [Lasken RS: Single-cell genomic sequencing using multiple displacement amplification, Curr Opin Microbiol 2007, 10:510-516]. Here, we focus on research strategies for putting these methods into practice in the laboratory setting. An immediate consequence of single-cell sequencing is that it provides an alternative to culturing organisms as a prerequisite for genomic sequencing. The microgram amounts of DNA required as template are amplified from a single bacterium by a method called multiple displacement amplification (MDA) avoiding the need to grow cells. The ability to sequence DNA from individual cells will likely have an immense impact on microbiology considering the vast numbers of novel organisms, which have been inaccessible unless culture-independent methods could be used. However, special approaches have been necessary to work with amplified DNA. MDA may not recover the entire genome from the single copy present in most bacteria. Also, some sequence rearrangements can occur during the DNA amplification reaction. Over the past two years many research groups have begun to use MDA, and some practical approaches to single-cell sequencing have been developed. We review the consensus that is emerging on optimum methods, reliability of amplified template, and the proper interpretation of 'composite' genomes which result from the necessity of combining data from several single-cell MDA reactions in order to complete the assembly. Preferred laboratory methods are considered on the basis of experience at several large sequencing centers where >70% of genomes are now often recovered from single cells. Methods are reviewed for preparation of bacterial fractions from environmental samples, single-cell isolation, DNA amplification by MDA, and DNA sequencing.

  2. Improving flavor metabolism of Saccharomyces cerevisiae by mixed culture with Bacillus licheniformis for Chinese Maotai-flavor liquor making.

    PubMed

    Meng, Xing; Wu, Qun; Wang, Li; Wang, Diqiang; Chen, Liangqiang; Xu, Yan

    2015-12-01

    Microbial interactions could impact the metabolic behavior of microbes involved in food fermentation, and therefore they are important for improving food quality. This study investigated the effect of Bacillus licheniformis, the dominant bacteria in the fermentation process of Chinese Maotai-flavor liquor, on the metabolic activity of Saccharomyces cerevisiae. Results indicated that S. cerevisiae inhibited the growth of B. licheniformis in all mixed culture systems and final viable cell count was lower than 20 cfu/mL. Although growth of S. cerevisiae was barely influenced by B. licheniformis, its metabolism was changed as initial inoculation ratio varied. The maximum ethanol productions were observed in S. cerevisiae and B. licheniformis at 10(6):10(7) and 10(6):10(8) ratios and have increased by 16.8 % compared with single culture of S. cerevisiae. According to flavor compounds, the culture ratio 10(6):10(6) showed the highest level of total concentrations of all different kinds of flavor compounds. Correlation analyses showed that 12 flavor compounds, including 4 fatty acids and their 2 corresponding esters, 1 terpene, and 5 aromatic compounds, that could only be produced by S. cerevisiae were significantly correlated with the initial inoculation amount of B. licheniformis. These metabolic changes in S. cerevisiae were not only a benefit for liquor aroma, but may also be related to its inhibition effect in mixed culture. This study could help to reveal the microbial interactions in Chinese liquor fermentation and provide guidance for optimal arrangement of mixed culture fermentation systems. PMID:26323612

  3. A Role for Programmed Cell Death in the Microbial Loop

    PubMed Central

    Durand, Pierre M.; Whitehead, Kenia; Baliga, Nitin S.

    2013-01-01

    The microbial loop is the conventional model by which nutrients and minerals are recycled in aquatic eco-systems. Biochemical pathways in different organisms become metabolically inter-connected such that nutrients are utilized, processed, released and re-utilized by others. The result is that unrelated individuals end up impacting each others' fitness directly through their metabolic activities. This study focused on the impact of programmed cell death (PCD) on a population's growth as well as its role in the exchange of carbon between two naturally co-occurring halophilic organisms. Flow cytometric, biochemical, 14C radioisotope tracing assays, and global transcriptomic analyses show that organic algal photosynthate released by Dunalliela salina cells undergoing PCD complements the nutritional needs of other non-PCD D. salina cells. This occurs in vitro in a carbon limited environment and enhances the growth of the population. In addition, a co-occurring heterotroph Halobacterium salinarum re-mineralizes the carbon providing elemental nutrients for the mixoheterotrophic chlorophyte. The significance of this is uncertain and the archaeon can also subsist entirely on the lysate of apoptotic algae. PCD is now well established in unicellular organisms; however its ecological relevance has been difficult to decipher. In this study we found that PCD in D. salina causes the release of organic nutrients such as glycerol, which can be used by others in the population as well as a co-occurring halophilic archaeon. H. salinarum also re-mineralizes the dissolved material promoting algal growth. PCD in D. salina was the mechanism for the flow of dissolved photosynthate between unrelated organisms. Ironically, programmed death plays a central role in an organism's own population growth and in the exchange of nutrients in the microbial loop. PMID:23667496

  4. Electricity generation from tetrathionate in microbial fuel cells by acidophiles.

    PubMed

    Sulonen, Mira L K; Kokko, Marika E; Lakaniemi, Aino-Maija; Puhakka, Jaakko A

    2015-03-01

    Inorganic sulfur compounds, such as tetrathionate, are often present in mining process and waste waters. The biodegradation of tetrathionate was studied under acidic conditions in aerobic batch cultivations and in anaerobic anodes of two-chamber flow-through microbial fuel cells (MFCs). All four cultures originating from biohydrometallurgical process waters from multimetal ore heap bioleaching oxidized tetrathionate aerobically at pH below 3 with sulfate as the main soluble metabolite. In addition, all cultures generated electricity from tetrathionate in MFCs at pH below 2.5 with ferric iron as the terminal cathodic electron acceptor. The maximum current and power densities during MFC operation and in the performance analysis were 79.6 mA m(-2) and 13.9 mW m(-2) and 433 mA m(-2) and 17.6 mW m(-2), respectively. However, the low coulombic efficiency (below 5%) indicates that most of the electrons were directed to other processes, such as aerobic oxidation of tetrathionate and unmeasured intermediates. The microbial community analysis revealed that the dominant species both in the anolyte and on the anode electrode surface of the MFCs were Acidithiobacillus spp. and Ferroplasma spp. This study provides a proof of concept that tetrathionate serves as electron donor for biological electricity production in the pH range of 1.2-2.5. PMID:25463232

  5. Stimulating sediment bioremediation with benthic microbial fuel cells.

    PubMed

    Li, Wen-Wei; Yu, Han-Qing

    2015-01-01

    Efficient and sustainable technologies for cleaning up of contaminated sediments are under urgent demand. Bioremediation by utilizing the natural metabolic activities of sediment-inhabited microorganisms has been widely accepted as a viable option, but the relatively low efficiency and poor controllability severely limite its application. Here, we bring out the concept that electrochemical approaches may be used as an efficient means to stimulate sediment bioremediation. Although still at the very beginning, benthic microbial fuel cells (BMFC) as a remediation technology show many potential benefits, such as accelerated decontamination, self-sustained operation, relatively easy deployment and control, and environmental benignity. The unique features of BMFC setup and operation also give rise to substantially different challenges compared to conventional MFCs. In this review, we present a critical overview on the characteristics, possible application niches, and state-of-the-art progress of this technology. Especially, the current limitations in respect of system design, electrode selection, microbial control and selection of deployment environment are discussed in details, and the needed future research endeavors to promote its practical application are highlighted. PMID:25560929

  6. New applications of carbon nanostructures in microbial fuel cells (MFC)

    NASA Astrophysics Data System (ADS)

    Kaca, W.; Żarnowiec, P.; Keczkowska, Justyna; Suchańska, M.; Czerwosz, E.; Kozłowski, M.

    2014-11-01

    In the studies presented we proposed a new application for nanocomposite carbon films (C-Pd). These films were evaluated as an anode material for Microbial Fuel Cells (MFCs) used for electrical current generation. The results of characterization of C-Pd films composed of carbon and palladium nanograins were obtained using the Physical Vapor Deposition (PVD) method. The film obtained by this method exhibits a multiphase structure composed of fullerene nanograins, amorphous carbon and palladium nanocrystals. Raman Spectroscopy (RS) and scanning electron microscopy (SEM) are used to characterize the chemical composition, morphology and topography of these films. We observed, for MFC with C-Pd anode, the highest electrochemical activity and maximal voltage density - 458 mV (20,8 mV/cm2) for Proteus mirabilis, 426 mV (19,4 mV/cm2) for Pseudomonas aeruginosa and 652 mV (29,6 mV/cm2) for sewage bacteria as the microbial catalyst.

  7. Flow cytometry and cell sorting of heterogeneous microbial populations: the importance of single-cell analyses.

    PubMed Central

    Davey, H M; Kell, D B

    1996-01-01

    The most fundamental questions such as whether a cell is alive, in the sense of being able to divide or to form a colony, may sometimes be very hard to answer, since even axenic microbial cultures are extremely heterogeneous. Analyses that seek to correlate such things as viability, which is a property of an individual cell, with macroscopic measurements of culture variables such as ATP content, respiratory activity, and so on, must inevitably fail. It is therefore necessary to make physiological measurements on individual cells. Flow cytometry is such a technique, which allows one to analyze cells rapidly and individually and permits the quantitative analysis of microbial heterogeneity. It therefore offers many advantages over conventional measurements for both routine and more exploratory analyses of microbial properties. While the technique has been widely applied to the study of mammalian cells, is use in microbiology has until recently been much more limited, largely because of the smaller size of microbes and the consequently smaller optical signals obtainable from them. Since these technical barriers no longer hold, flow cytometry with appropriate stains has been used for the rapid discrimination and identification of microbial cells, for the rapid assessment of viability and of the heterogeneous distributions of a wealth of other more detailed physiological properties, for the analysis of antimicrobial drug-cell interactions, and for the isolation of high-yielding strains of biotechnological interest. Flow cytometric analyses provide an abundance of multivariate data, and special methods have been devised to exploit these. Ongoing advances mean that modern flow cytometers may now be used by nonspecialists to effect a renaissance in our understanding of microbial heterogeneity. PMID:8987359

  8. Molecular characterization of cell cycle gene CDC7 from Saccharomyces cerevisiae.

    PubMed Central

    Patterson, M; Sclafani, R A; Fangman, W L; Rosamond, J

    1986-01-01

    The product of the CDC7 gene of Saccharomyces cerevisiae appears to have multiple roles in cellular physiology. It is required for the initiation of mitotic DNA synthesis. While it is not required for the initiation of meiotic DNA replication, it is necessary for genetic recombination during meiosis and for the formation of ascospores. It has also been implicated in an error-prone DNA repair pathway. Plasmids capable of complementing temperature-sensitive cdc7 mutations were isolated from libraries of yeast genomic DNA in the multicopy plasmid vectors YRp7 and YEp24. The complementing activity was localized within a 3.0-kilobase genomic DNA fragment. Genetic studies that included integration of the genomic insert at or near the CDC7 locus and marker rescue of four cdc7 alleles proved that the cloned fragment contains the yeast chromosomal CDC7 gene. The RNA transcript of CDC7 is about 1,700 nucleotides. Analysis of the nucleotide sequence of a 2.1-kilobase region of the cloned fragment revealed the presence of an open reading frame of 1,521 nucleotides that is presumed to encode the CDC7 protein. Depending on which of two possible ATG codons initiates translation, the calculated size of the CDC7 protein is 58.2 or 56 kilodaltons. Comparison of the predicted amino acid sequence of the CDC7 gene product with other known protein sequences suggests that CDC7 encodes a protein kinase. Images PMID:3537706

  9. A comparative evaluation of different types of microbial electrolysis desalination cells for malic acid production.

    PubMed

    Liu, Guangli; Zhou, Ying; Luo, Haiping; Cheng, Xing; Zhang, Renduo; Teng, Wenkai

    2015-12-01

    The aim of this study was to investigate different microbial electrolysis desalination cells for malic acid production. The systems included microbial electrolysis desalination and chemical-production cell (MEDCC), microbial electrolysis desalination cell (MEDC) with bipolar membrane and anion exchange membrane (BP-A MEDC), MEDC with bipolar membrane and cation exchange membrane (BP-C MEDC), and modified microbial desalination cell (M-MDC). The microbial electrolysis desalination cells performed differently in terms of malic acid production and energy consumption. The MEDCC performed best with the highest malic acid production rate (18.4 ± 0.6 mmol/Lh) and the lowest energy consumption (0.35 ± 0.14 kWh/kg). The best performance of MEDCC was attributable to the neutral pH condition in the anode chamber, the lowest internal resistance, and the highest Geobacter percentage of the anode biofilm population among all the reactors. PMID:26367771

  10. Optical manipulation of Saccharomyces cerevisiae cells reveals that green light protection against UV irradiation is favored by low Ca2+ and requires intact UPR pathway.

    PubMed

    Farcasanu, Ileana C; Mitrica, Radu; Cristache, Ligia; Nicolau, Ioana; Ruta, Lavinia L; Paslaru, Liliana; Comorosan, Sorin

    2013-11-01

    Optical manipulation of Saccharomyces cerevisiae cells with high density green photons conferred protection against the deleterious effects of UV radiation. Combining chemical screening with UV irradiation of yeast cells, it was noted that the high density green photons relied on the presence of intact unfolded protein response (UPR) pathway to exert their protective effect and that the low Ca(2+) conditions boosted the effect. UPR chemical inducers tunicamycin, dithiotreitol and calcium chelators augmented the green light effect in a synergic action against UV-induced damage. Photo-manipulation of cells was a critical factor since the maximum protection was achieved only when cells were pre-exposed to green light. PMID:24056073

  11. Pinostrobin from Boesenbergia pandurata is an inhibitor of Ca2+-signal-mediated cell-cycle regulation in the yeast Saccharomyces cerevisiae.

    PubMed

    Wangkangwan, Wachirasak; Boonkerd, Saipin; Chavasiri, Warinthorn; Sukapirom, Kasama; Pattanapanyasat, Kovit; Kongkathip, Ngampong; Miyakawa, Tokichi; Yompakdee, Chulee

    2009-07-01

    Upon searching plant extracts for inhibitors of the Ca(2+) signaling pathway using the zds1Delta-yeast proliferation based assay, a crude rhizome extract of Boesenbergia pandurata was found to be strongly positive, and from this extract pinostrobin, alpinetin, and pinocembrin chalcone were isolated as active components. Further biochemical experiments confirmed that pinostrobin possesses inhibitory activity on the Ca(2+) signals involved in the control of G2/M phase cell cycle progression in Saccharomyces cerevisiae. PMID:19584530

  12. Cell surface engineering of Saccharomyces cerevisiae combined with membrane separation technology for xylitol production from rice straw hydrolysate.

    PubMed

    Guirimand, Gregory; Sasaki, Kengo; Inokuma, Kentaro; Bamba, Takahiro; Hasunuma, Tomohisa; Kondo, Akihiko

    2016-04-01

    Xylitol, a value-added polyol deriving from D-xylose, is widely used in both the food and pharmaceutical industries. Despite extensive studies aiming to streamline the production of xylitol, the manufacturing cost of this product remains high while demand is constantly growing worldwide. Biotechnological production of xylitol from lignocellulosic waste may constitute an advantageous and sustainable option to address this issue. However, to date, there have been few reports of biomass conversion to xylitol. In the present study, xylitol was directly produced from rice straw hydrolysate using a recombinant Saccharomyces cerevisiae YPH499 strain expressing cytosolic xylose reductase (XR), along with β-glucosidase (BGL), xylosidase (XYL), and xylanase (XYN) enzymes (co-)displayed on the cell surface; xylitol production by this strain did not require addition of any commercial enzymes. All of these enzymes contributed to the consolidated bioprocessing (CBP) of the lignocellulosic hydrolysate to xylitol to produce 5.8 g/L xylitol with 79.5 % of theoretical yield from xylose contained in the biomass. Furthermore, nanofiltration of the rice straw hydrolysate provided removal of fermentation inhibitors while simultaneously increasing sugar concentrations, facilitating high concentration xylitol production (37.9 g/L) in the CBP. This study is the first report (to our knowledge) of the combination of cell surface engineering approach and membrane separation technology for xylitol production, which could be extended to further industrial applications. PMID:26631184

  13. Predicting complex phenotype-genotype interactions to enable yeast engineering: Saccharomyces cerevisiae as a model organism and a cell factory.

    PubMed

    Dikicioglu, Duygu; Pir, Pınar; Oliver, Stephen G

    2013-09-01

    There is an increasing use of systems biology approaches in both "red" and "white" biotechnology in order to enable medical, medicinal, and industrial applications. The intricate links between genotype and phenotype may be explained through the use of the tools developed in systems biology, synthetic biology, and evolutionary engineering. Biomedical and biotechnological research are among the fields that could benefit most from the elucidation of this complex relationship. Researchers have studied fitness extensively to explain the phenotypic impacts of genetic variations. This elaborate network of dependencies and relationships so revealed are further complicated by the influence of environmental effects that present major challenges to our achieving an understanding of the cellular mechanisms leading to healthy or diseased phenotypes or optimized production yields. An improved comprehension of complex genotype-phenotype interactions and their accurate prediction should enable us to more effectively engineer yeast as a cell factory and to use it as a living model of human or pathogen cells in intelligent screens for new drugs. This review presents different methods and approaches undertaken toward improving our understanding and prediction of the growth phenotype of the yeast Saccharomyces cerevisiae as both a model and a production organism. PMID:24031036

  14. Long-term operation of manure-microbial fuel cell.

    PubMed

    Zhang, Guodong; Zhao, Qingliang; Jiao, Yan; Lee, Duu-Jong

    2015-03-01

    Microbial fuel cell (MFC) is applied to produce electricity using dairy manure as a fuel. Since the way MFC utilizes manure as a fuel and the long-term operation stability of manure-MFC remains unclear, this study examined the evolution of dissolved organic matter (DOM) in anodic chamber and power generation by MFC in a 171days test. The tested MFC can produce electricity over the entire testing period by single feed of manure, with stable power output and total chemical oxygen demand (TCOD) removal rate in the period of day 30-140. The hydrophobic acid (HPO-A) and hydrophilic (HPI) fractions of manure were the principal components of anolyte DOM, with the concentrations of both being reduced over MFC operation. The degradable organic matters were converted to compounds with high aromaticity. PMID:25603729

  15. Experimental study of the microbial fuel cell internal resistance

    NASA Astrophysics Data System (ADS)

    Zhang, Pei-Yuan; Liu, Zhong-Liang

    The internal resistance, including activation loss internal resistance (AIR), ohmic loss internal resistance (OIR) and concentration loss internal resistance (CIR), is an important parameter that determines the performance of microbial fuel cells (MFCs). The experimental investigations were completed to estimate the contributions of these three components to the internal resistance. The internal resistance is found to vary with electric current, although it is almost a constant for the current is within a certain region. The largest component of the internal resistance is CIR except for small currents. The AIR decreases quickly for small current and reduces its decreasing rate as the current increases and approaches to a constant. The OIR is constant over the whole current range. The experiments also disclose that increasing the limiting current and reducing the concentration loss are both important for improving the MFC performance.

  16. Microfluidic microbial fuel cells: from membrane to membrane free

    NASA Astrophysics Data System (ADS)

    Yang, Yang; Ye, Dingding; Li, Jun; Zhu, Xun; Liao, Qiang; Zhang, Biao

    2016-08-01

    Microfluidic microbial fuel cells (MMFCs) are small carbon-neutral devices that use self-organized bacteria to degrade organic substrates and harness energy from the waste water. Conventional MMFCs have made great strides in the past decade and have overcome some limitations, such as high capital costs and low energy output. A co-laminar flow MFC has been first proposed in 2011 with the potential to be an attractively power source to niche applications. Co-laminar MFCs typically operate without any physical membranes separating the reactants, and bacterial ecosystems can be easily manipulated by regulating the inlet conditions. This paper highlights recent accomplishments in the development of co-laminar MFCs, emphasizing basic principles, mass transport and fluid dynamics including boundary layer theory, entrance conditions and mixing zone issues. Furthermore, the development of current techniques, major challenges and the potential research directions are discussed.

  17. Fade to Green: A Biodegradable Stack of Microbial Fuel Cells.

    PubMed

    Winfield, Jonathan; Chambers, Lily D; Rossiter, Jonathan; Stinchcombe, Andrew; Walter, X Alexis; Greenman, John; Ieropoulos, Ioannis

    2015-08-24

    The focus of this study is the development of biodegradable microbial fuel cells (MFCs) able to produce useful power. Reactors with an 8 mL chamber volume were designed using all biodegradable products: polylactic acid for the frames, natural rubber as the cation-exchange membrane and egg-based, open-to-air cathodes coated with a lanolin gas diffusion layer. Forty MFCs were operated in various configurations. When fed with urine, the biodegradable stack was able to power appliances and was still operational after six months. One useful application for this truly sustainable MFC technology includes onboard power supplies for biodegradable robotic systems. After operation in remote ecological locations, these could degrade harmlessly into the surroundings to leave no trace when the mission is complete. PMID:26212495

  18. Experimenting with microbial fuel cells for powering implanted biomedical devices.

    PubMed

    Roxby, Daniel N; Nham Tran; Pak-Lam Yu; Nguyen, Hung T

    2015-08-01

    Microbial Fuel Cell (MFC) technology has the ability to directly convert sugar into electricity by using bacteria. Such a technology could be useful for powering implanted biomedical devices that require a surgery to replace their batteries every couple of years. In steps towards this, parameters such as electrode configuration, inoculation size, stirring of the MFC and single versus dual chamber reactor configuration were tested for their effect on MFC power output. Results indicate that a Top-Bottom electrode configuration, stirring and larger amounts of bacteria in single chamber MFCs, and smaller amounts of bacteria in dual chamber MFCs give increased power outputs. Finally, overall dual chamber MFCs give several fold larger MFC power outputs. PMID:26736845

  19. Performance of Denitrifying Microbial Fuel Cell with Biocathode over Nitrite

    PubMed Central

    Zhao, Huimin; Zhao, Jianqiang; Li, Fenghai; Li, Xiaoling

    2016-01-01

    Microbial fuel cell (MFC) with nitrite as an electron acceptor in cathode provided a new technology for nitrogen removal and electricity production simultaneously. The influences of influent nitrite concentration and external resistance on the performance of denitrifying MFC were investigated. The optimal effectiveness were obtained with the maximum total nitrogen (TN) removal rate of 54.80 ± 0.01 g m−3 d−1. It would be rather desirable for the TN removal than electricity generation at lower external resistance. Denaturing gradient gel electrophoresis suggested that Proteobacteria was the predominant phylum, accounting for 35.72%. Thiobacillus and Afipia might benefit to nitrite removal. The presence of nitrifying Devosia indicated that nitrite was oxidized to nitrate via a biochemical mechanism in the cathode. Ignavibacterium and Anaerolineaceae was found in the cathode as a heterotrophic bacterium with sodium acetate as substrate, which illustrated that sodium acetate in anode was likely permeated through proton exchange membrane to the cathode. PMID:27047462

  20. Microbial fuel cell treatment of ethanol fermentation process water

    SciTech Connect

    Borole, Abhijeet P.

    2012-06-05

    The present invention relates to a method for removing inhibitor compounds from a cellulosic biomass-to-ethanol process which includes a pretreatment step of raw cellulosic biomass material and the production of fermentation process water after production and removal of ethanol from a fermentation step, the method comprising contacting said fermentation process water with an anode of a microbial fuel cell, said anode containing microbes thereon which oxidatively degrade one or more of said inhibitor compounds while producing electrical energy or hydrogen from said oxidative degradation, and wherein said anode is in electrical communication with a cathode, and a porous material (such as a porous or cation-permeable membrane) separates said anode and cathode.

  1. Autotrophic nitrite removal in the cathode of microbial fuel cells.

    PubMed

    Puig, Sebastià; Serra, Marc; Vilar-Sanz, Ariadna; Cabré, Marina; Bañeras, Lluís; Colprim, Jesús; Balaguer, M Dolors

    2011-03-01

    Nitrification to nitrite (nitritation process) followed by reduction to dinitrogen gas decreases the energy demand and the carbon requirements of the overall process of nitrogen removal. This work studies autotrophic nitrite removal in the cathode of microbial fuel cells (MFCs). Special attention was paid to determining whether nitrite is used as the electron acceptor by exoelectrogenic bacteria (biologic reaction) or by graphite electrodes (abiotic reaction). The results demonstrated that, after a nitrate pulse at the cathode, nitrite was initially accumulated; subsequently, nitrite was removed. Nitrite and nitrate can be used interchangeably as an electron acceptor by exoelectrogenic bacteria for nitrogen reduction from wastewater while producing bioelectricity. However, if oxygen is present in the cathode chamber, nitrite is oxidised via biological or electrochemical processes. The identification of a dominant bacterial member similar to Oligotropha carboxidovorans confirms that autotrophic denitrification is the main metabolism mechanism in the cathode of an MFC. PMID:21262566

  2. Scaling up microbial fuel cells and other bioelectrochemical systems.

    PubMed

    Logan, Bruce E

    2010-02-01

    Scientific research has advanced on different microbial fuel cell (MFC) technologies in the laboratory at an amazing pace, with power densities having reached over 1 kW/m(3) (reactor volume) and to 6.9 W/m(2) (anode area) under optimal conditions. The main challenge is to bring these technologies out of the laboratory and engineer practical systems for bioenergy production at larger scales. Recent advances in new types of electrodes, a better understanding of the impact of membranes and separators on performance of these systems, and results from several new pilot-scale tests are all good indicators that commercialization of the technology could be possible within a few years. Some of the newest advances and future challenges are reviewed here with respect to practical applications of these MFCs for renewable energy production and other applications. PMID:20013119

  3. Carbon nanotube modification of microbial fuel cell electrodes.

    PubMed

    Yazdi, Alireza Ahmadian; D'Angelo, Lorenzo; Omer, Nada; Windiasti, Gracia; Lu, Xiaonan; Xu, Jie

    2016-11-15

    The use of carbon nanotubes (CNTs) for energy harvesting devices is preferable due to their unique mechanical, thermal, and electrical properties. On the other hand, microbial fuel cells (MFCs) are promising devices to recover carbon-neutral energy from the organic matters, and have been hindered with major setbacks towards commercialization. Nanoengineered CNT-based materials show remarkable electrochemical properties, and therefore have provided routes towards highly effective modification of MFC compartments to ultimately reach the theoretical limits of biomass energy recovery, low-cost power production, and thus the commercialization of MFCs. Moreover, these CNT-based composites offer significant flexibility in the design of MFCs that enable their use for a broad spectrum of applications ranging from scaled-up power generation to medically related devices. This article reviews the recent advances in the modification of MFCs using CNTs and CNT-based composites, and the extent to which each modification route impacts MFC power and current generation. PMID:27213269

  4. Microbial desalination cell with capacitive adsorption for ion migration control.

    PubMed

    Forrestal, Casey; Xu, Pei; Jenkins, Peter E; Ren, Zhiyong

    2012-09-01

    A new microbial desalination cell with capacitive adsorption capability (cMDC) was developed to solve the ion migration problem facing current MDC systems. Traditional MDCs remove salts by transferring ions to the anode and cathode chambers, which may prohibit wastewater beneficial reuse due to increased salinity. The cMDC uses adsorptive activated carbon cloth (ACC) as the electrodes and utilizes the formed capacitive double layers for electrochemical ion adsorption. The cMDC removed an average of 69.4% of the salt from the desalination chamber through electrode adsorption during one batch cycle, and it did not add salts to the anode or cathode chamber. It was estimated that 61-82.2mg of total dissolved solids (TDS) was adsorbed to 1g of ACC electrode. The cMDC provides a new approach for salt management, organic removal, and energy production. Further studies will be conducted to optimize reactor configuration and achieve in situ electrode regeneration. PMID:22784594

  5. Genome-wide analysis of longevity in nutrient-deprived Saccharomyces cerevisiae reveals importance of recycling in maintaining cell viability.

    PubMed

    Davey, Hazel M; Cross, Emma J M; Davey, Christopher L; Gkargkas, Konstantinos; Delneri, Daniela; Hoyle, David C; Oliver, Stephen G; Kell, Douglas B; Griffith, Gareth W

    2012-05-01

    Although typically cosseted in the laboratory with constant temperatures and plentiful nutrients, microbes are frequently exposed to much more stressful conditions in their natural environments where survival and competitive fitness depend upon both growth rate when conditions are favourable and on persistence in a viable and recoverable state when they are not. In order to determine the role of genetic heterogeneity in environmental fitness we present a novel approach that combines the power of fluorescence-activated cell sorting with barcode microarray analysis and apply this to determining the importance of every gene in the Saccharomyces cerevisiae genome in a high-throughput, genome-wide fitness screen. We have grown > 6000 heterozygous mutants together and exposed them to a starvation stress before using fluorescence-activated cell sorting to identify and isolate those individual cells that have not survived the stress applied. Barcode array analysis of the sorted and total populations reveals the importance of cellular recycling mechanisms (autophagy, pexophagy and ribosome breakdown) in maintaining cell viability during starvation and provides compelling evidence for an important role for fatty acid degradation in maintaining viability. In addition, we have developed a semi-batch fermentor system that is a more realistic model of environmental fitness than either batch or chemostat culture. Barcode array analysis revealed that arginine biosynthesis was important for fitness in semi-batch culture and modelling of this regime showed that rapid emergence from lag phase led to greatly increased fitness. One hundred and twenty-five strains with deletions in unclassified proteins were identified as being over-represented in the sorted fraction, while 27 unclassified proteins caused a haploinsufficient phenotype in semi-batch culture. These methods thus provide a screen to identifying other genes and pathways that have a role in maintaining cell viability. PMID

  6. Critical analysis of the maximum non inhibitory concentration (MNIC) method in quantifying sub-lethal injury in Saccharomyces cerevisiae cells exposed to either thermal or pulsed electric field treatments.

    PubMed

    Kethireddy, V; Oey, I; Jowett, Tim; Bremer, P

    2016-09-16

    Sub-lethal injury within a microbial population, due to processing treatments or environmental stress, is often assessed as the difference in the number of cells recovered on non-selective media compared to numbers recovered on a "selective media" containing a predetermined maximum non-inhibitory concentration (MNIC) of a selective agent. However, as knowledge of cell metabolic response to injury, population diversity and dynamics increased, the rationale behind the conventional approach of quantifying sub-lethal injury must be scrutinized further. This study reassessed the methodology used to quantify sub-lethal injury for Saccharomyces cerevisiae cells (≈ 4.75 Log CFU/mL) exposed to either a mild thermal (45°C for 0, 10 and 20min) or a mild pulsed electric field treatment (field strengths of 8.0-9.0kV/cm and energy levels of 8, 14 and 21kJ/kg). Treated cells were plated onto either Yeast Malt agar (YM) or YM containing NaCl, as a selective agent at 5-15% in 1% increments. The impact of sub-lethal stress due to initial processing, the stress due to selective agents in the plating media, and the subsequent variation of inhibition following the treatments was assessed based on the CFU count (cell numbers). ANOVA and a generalised least squares model indicated significant effects of media, treatments, and their interaction effects (P<0.05) on cell numbers. It was shown that the concentration of the selective agent used dictated the extent of sub-lethal injury recorded owing to the interaction effects of the selective component (NaCl) in the recovery media. Our findings highlight a potential common misunderstanding on how culture conditions impact on sub-lethal injury. Interestingly for S. cerevisiae cells the number of cells recovered at different NaCl concentrations in the media appears to provide valuable information about the mode of injury, the comparative efficacy of different processing regimes and the inherent degree of resistance within a population. This

  7. Microbial fuel cells for clogging assessment in constructed wetlands.

    PubMed

    Corbella, Clara; García, Joan; Puigagut, Jaume

    2016-11-01

    Clogging in HSSF CW may result in a reduction of system's life-span or treatment efficiency. Current available techniques to assess the degree of clogging in HSSF CW are time consuming and cannot be applied on a continuous basis. Main objective of this work was to assess the potential applicability of microbial fuel cells for continuous clogging assessment in HSSF CW. To this aim, two replicates of a membrane-less microbial fuel cell (MFC) were built up and operated under laboratory conditions for five weeks. The MFC anode was gravel-based to simulate the filter media of HSSF CW. MFC were weekly loaded with sludge that had been accumulating for several years in a pilot HSSF CW treating domestic wastewater. Sludge loading ranged from ca. 20kgTS·m(-3)CW·year(-1) at the beginning of the study period up to ca. 250kgTS·m(-3)CW·year(-1) at the end of the study period. Sludge loading applied resulted in sludge accumulated within the MFC equivalent to a clogging degree ranging from 0.2years (ca. 0.5kgTS·m(-3)CW) to ca. 5years (ca. 10kgTS·m(-3)CW). Results showed that the electric charge was negatively correlated to the amount of sludge accumulated (degree of clogging). Electron transference (expressed as electric charge) almost ceased when accumulated sludge within the MFC was equivalent to ca. 5years of clogging (ca. 10kgTS·m(-3)CW). This result suggests that, although longer study periods under more realistic conditions shall be further performed, HSSF CW operated as a MFC has great potential for clogging assessment. PMID:27392579

  8. A new method for water desalination using microbial desalination cells.

    PubMed

    Cao, Xiaoxin; Huang, Xia; Liang, Peng; Xiao, Kang; Zhou, Yingjun; Zhang, Xiaoyuan; Logan, Bruce E

    2009-09-15

    Current water desalination techniques are energy intensive and some use membranes operated at high pressures. It is shown here that water desalination can be accomplished without electrical energy input or high water pressure by using a source of organic matter as the fuel to desalinate water. A microbial fuel cell was modified by placing two membranes between the anode and cathode, creating a middle chamber for water desalination between the membranes. An anion exchange membrane was placed adjacent to the anode, and a cation exchange membrane was positioned next to the cathode. When current was produced by bacteria on the anode, ionic species in the middle chamber were transferred into the two electrode chambers, desalinating the water in the middle chamber. Proof-of-concept experiments for this approach, using what we call a microbial desalination cell (MDC), was demonstrated using water at different initial salt concentrations (5, 20, and 35 g/L) with acetate used as the substrate for the bacteria. The MDC produced a maximum of 2 W/m2 (31 W/m3) while at the same time removing about 90% of the salt in a single desalination cycle. As the salt was removed from the middle chamber the ohmic resistance of the MDC (measured using electrochemical impedance spectroscopy) increased from 25 Omega to 970 Omega at the end of the cycle. This increased resistance was reflected by a continuous decrease in the voltage produced over the cycle. These results demonstrate for the first time the possibility for a new method for water desalination and power production that uses only a source of biodegradable organic matter and bacteria. PMID:19806756

  9. Microbial analysis of anodic biofilm in a microbial fuel cell using slaughterhouse wastewater.

    PubMed

    Katuri, Krishna P; Enright, Ann-Marie; O'Flaherty, Vincent; Leech, Dónal

    2012-10-01

    The ability of dual-chambered microbial fuel cell, fed with slaughterhouse wastewater with an anaerobic mixed-sludge as initial source of bacteria, to generate power is investigated. MFC voltage generation across a fixed 100 Ω load indicates power generation capability, with power production correlated to changes in anolyte VFA content. A maximum MFC power density of 578 mW/m(2) is obtained for an MFC developed under 100 Ω load, compared to a maximum power density of 277 mW/m(2) for an MFC developed under higher resistance (1 MΩ) control conditions. Voltammetry of the biofilm developed under 100 Ω load displays a current-voltage signal indicative of bioelectrocatalytic oxidation of feed at a potential of -0.35 V vs. Ag/AgCl, compared to negligible signals for biofilms developed under control conditions. Denaturing gradient gel electrophoresis of PCR amplified 16S rRNA gene fragments reveals that the anodic bacterial communities in reactors operated under 100 Ω load result in communities of lower diversity than for the control condition, with Geovibrio ferrireducens dominant in the anodic biofilm community. These results indicate that in MFC reactors, functionally stable electroactive bacteria are enriched under 100 Ω load compared to high resistance control conditions, and were able to sustain higher power in MFCs. PMID:22226620

  10. Microbial community analysis of anodes from sediment microbial fuel cells powered by rhizodeposits of living rice plants.

    PubMed

    De Schamphelaire, Liesje; Cabezas, Angela; Marzorati, Massimo; Friedrich, Michael W; Boon, Nico; Verstraete, Willy

    2010-03-01

    By placing the anode of a sediment microbial fuel cell (SMFC) in the rhizosphere of a rice plant, root-excreted rhizodeposits can be microbially oxidized with concomitant current generation. Here, various molecular techniques were used to characterize the composition of bacterial and archaeal communities on such anodes, as influenced by electrical circuitry, sediment matrix, and the presence of plants. Closed-circuit anodes in potting soil were enriched with Desulfobulbus-like species, members of the family Geobacteraceae, and as yet uncultured representatives of the domain Archaea. PMID:20097806

  11. High-cell-density fermentation of Saccharomyces cerevisiae for the optimisation of mead production.

    PubMed

    Pereira, A P; Mendes-Ferreira, A; Oliveira, J M; Estevinho, L M; Mendes-Faia, A

    2013-02-01

    Mead is a traditional drink that contains 8%-18% (v/v) of ethanol, resulting from the alcoholic fermentation of diluted honey by yeasts. Mead fermentation is a time-consuming process and the quality of the final product is highly variable. Therefore, the present investigation had two main objectives: first, to determine the adequate inoculum size of two commercial wine-making strains of Saccharomyces cerevisiae for the optimisation of mead fermentation; and second, to determine if an increase in yeast pitching rates in batch fermentations altered the resulting aroma profiles. Minor differences were detected in the growth kinetics between the two strains at the lowest pitching rate. With increasing pitching rates net growth of the strain ICV D47 progressively decreased, whereas for the QA23 the increasing inoculum size had no influence on its net growth. The time required to reach the same stage of fermentation ranged from 24 to 96 h depending on the inoculum size. The final aroma composition was dependent on the yeast strain and inoculum size. Fourteen of the twenty-seven volatile compounds quantified could contribute to mead aroma and flavour because their concentrations rose above their respective thresholds. The formation of these compounds was particularly pronounced at low pitching rates, except in mead fermented by strain ICV D47, at 10(6) CFUs/mL. The esters isoamyl acetate, ethyl octanoate and ethyl hexanoate were the major powerful odourants found in the meads. The results obtained in this study demonstrate that yeast strain and inoculum size can favourably impact mead's flavour and aroma profiles. PMID:23122509

  12. Induction of a Mitosis Delay and Cell Lysis by High-Level Secretion of Mouse α-Amylase from Saccharomyces cerevisiae

    PubMed Central

    Wang, Bi-Dar; Kuo, Tsong-Teh

    2001-01-01

    Some foreign proteins are produced in yeast in a cell cycle-dependent manner, but the cause of the cell cycle dependency is unknown. In this study, we found that Saccharomyces cerevisiae cells secreting high levels of mouse α-amylase have elongated buds and are delayed in cell cycle completion in mitosis. The delayed cell mitosis suggests that critical events during exit from mitosis might be disturbed. We found that the activities of PP2A (protein phosphatase 2A) and MPF (maturation-promoting factor) were reduced in α-amylase-oversecreting cells and that these cells showed a reduced level of assembly checkpoint protein Cdc55, compared to the accumulation in wild-type cells. MPF inactivation is due to inhibitory phosphorylation on Cdc28, as a cdc28 mutant which lacks an inhibitory phosphorylation site on Cdc28 prevents MPF inactivation and prevents the defective bud morphology induced by overproduction of α-amylase. Our data also suggest that high levels of α-amylase may downregulate PPH22, leading to cell lysis. In conclusion, overproduction of heterologous α-amylase in S. cerevisiae results in a negative regulation of PP2A, which causes mitotic delay and leads to cell lysis. PMID:11472949

  13. Type 2C protein phosphatase Ptc6 participates in activation of the Slt2-mediated cell wall integrity pathway in Saccharomyces cerevisiae.

    PubMed

    Sharmin, Dilruba; Sasano, Yu; Sugiyama, Minetaka; Harashima, Satoshi

    2015-04-01

    The phosphorylation status of cellular proteins results from an equilibrium between the activities of protein kinases and protein phosphatases (PPases). Reversible protein phosphorylation is an important aspect of signal transduction that regulate many biological processes in eukaryotic cells. The Saccharomyces cerevisiae genome encodes 40 PPases, including seven members of the protein phosphatase 2C subfamily (PTC1 to PTC7). In contrast to other PPases, the cellular roles of PTCs have not been investigated in detail. Here, we sought to determine the cellular role of PTC6 in S. cerevisiae with disruption of PTC genes. We found that cells with Δptc6 disruption were tolerant to the cell wall-damaging agents Congo red (CR) and calcofluor white (CFW); however, cells with simultaneous disruption of PTC1 and PTC6 were very sensitive to these agents. Thus, simultaneous disruption of PTC1 and PTC6 gave a synergistic response to cell wall damaging agents. The level of phosphorylated Slt2 increased significantly after CR treatment in Δptc1 cells and more so in Δptc1Δptc6 cells; therefore, deletion of PTC6 enhanced Slt2 phosphorylation in the Δptc1 disruptant. The level of transcription of KDX1 upon exposure to CR increased to a greater extent in the Δptc1Δptc6 double disruptant than the Δptc1 single disruptant. The Δptc1Δptc6 double disruptant cells showed normal vacuole formation under standard growth conditions, but fragmented vacuoles were present in the presence of CR or CFW. Our analyses indicate that S. cerevisiae PTC6 participates in the negative regulation of Slt2 phosphorylation and vacuole morphogenesis under cell wall stress conditions. PMID:25449759

  14. From microbial fuel cell (MFC) to microbial electrochemical snorkel (MES): maximizing chemical oxygen demand (COD) removal from wastewater.

    PubMed

    Erable, Benjamin; Etcheverry, Luc; Bergel, Alain

    2011-03-01

    The paper introduces the concept of the microbial electrochemical snorkel (MES), a simplified design of a "short-circuited" microbial fuel cell (MFC). The MES cannot provide current but it is optimized for wastewater treatment. An electrochemically active biofilm (EAB) was grown on graphite felt under constant polarization in an urban wastewater. Controlling the electrode potential and inoculating the bioreactor with a suspension of an established EAB improved the performance and the reproducibility of the anodes. Anodes, colonized by an EAB were tested for the chemical oxygen demand (COD) removal from urban wastewater using a variety of bio-electrochemical processes (microbial electrolysis, MFC, MES). The MES technology, as well as a short-circuited MFC, led to a COD removal 57% higher than a 1000 Ω-connected MFC, confirming the potential for wastewater treatment. PMID:21409654

  15. Process intensification through microbial strain evolution: mixed glucose-xylose fermentation in wheat straw hydrolyzates by three generations of recombinant Saccharomyces cerevisiae

    PubMed Central

    2014-01-01

    Background Lignocellulose hydrolyzates present difficult substrates for ethanol production by the most commonly applied microorganism in the fermentation industries, Saccharomyces cerevisiae. High resistance towards inhibitors released during pretreatment and hydrolysis of the feedstock as well as efficient utilization of hexose and pentose sugars constitute major challenges in the development of S. cerevisiae strains for biomass-to-ethanol processes. Metabolic engineering and laboratory evolution are applied, alone and in combination, to adduce desired strain properties. However, physiological requirements for robust performance of S. cerevisiae in the conversion of lignocellulose hydrolyzates are not well understood. The herein presented S. cerevisiae strains IBB10A02 and IBB10B05 are descendants of strain BP10001, which was previously derived from the widely used strain CEN.PK 113-5D through introduction of a largely redox-neutral oxidoreductive xylose assimilation pathway. The IBB strains were obtained by a two-step laboratory evolution that selected for fast xylose fermentation in combination with anaerobic growth before (IBB10A02) and after adaption in repeated xylose fermentations (IBB10B05). Enzymatic hydrolyzates were prepared from up to 15% dry mass pretreated (steam explosion) wheat straw and contained glucose and xylose in a mass ratio of approximately 2. Results With all strains, yield coefficients based on total sugar consumed were high for ethanol (0.39 to 0.40 g/g) and notably low for fermentation by-products (glycerol: ≤0.10 g/g; xylitol: ≤0.08 g/g; acetate: 0.04 g/g). In contrast to the specific glucose utilization rate that was similar for all strains (qGlucose ≈ 2.9 g/gcell dry weight (CDW)/h), the xylose consumption rate was enhanced by a factor of 11.5 (IBB10A02; qXylose = 0.23 g/gCDW/h) and 17.5 (IBB10B05; qXylose = 0.35 g/gCDW/h) as compared to the qXylose of the non-evolved strain BP10001. In xylose-supplemented (50

  16. Inhibition of microbial growth on air cathodes of single chamber microbial fuel cells by incorporating enrofloxacin into the catalyst layer.

    PubMed

    Liu, Weifeng; Cheng, Shaoan; Sun, Dan; Huang, Haobin; Chen, Jie; Cen, Kefa

    2015-10-15

    The inevitable growth of aerobic bacteria on the surface of air cathodes is an important factor reducing the performance stability of air cathode single-chamber membrane-free microbial fuel cells (MFCs). Thus searching for effective methods to inhibit the cathodic microbial growth is critical for the practical application of MFCs. In this study, enrofloxacin (ENR), a broad spectrum fluoroquinolone antibiotic, was incorporated into the catalyst layer of activated carbon air cathodes (ACACs) to inhibit the cathodic microbial growth. The biomass content on ACACs was substantially reduced by 60.2% with ENR treatment after 91 days of MFCs operation. As a result of the inhibited microbial growth, the oxygen reduction catalytic performance of the ENR treated ACACs was much stable compared to the fast performance decline of the untreated control. Consequently, a quite stable electricity production was obtained for the MFCs with the ENR treated ACACs, in contrast with a 22.5% decrease in maximum power density of the MFCs with the untreated cathode. ENR treatment of ACACs showed minimal effects on the anode performance. These results indicate that incorporating antibiotics into ACACs should be a simple and effective strategy to inhibit the microbial growth and improve the long-term stability of the performance of air cathode and the electricity production of MFCs. PMID:25957076

  17. Single cell synchrotron FT-IR microspectroscopy reveals a link between neutral lipid and storage carbohydrate fluxes in S. cerevisiae.

    PubMed

    Jamme, Frédéric; Vindigni, Jean-David; Méchin, Valérie; Cherifi, Tamazight; Chardot, Thierry; Froissard, Marine

    2013-01-01

    In most organisms, storage lipids are packaged into specialized structures called lipid droplets. These contain a core of neutral lipids surrounded by a monolayer of phospholipids, and various proteins which vary depending on the species. Hydrophobic structural proteins stabilize the interface between the lipid core and aqueous cellular environment (perilipin family of proteins, apolipoproteins, oleosins). We developed a genetic approach using heterologous expression in Saccharomyces cerevisiae of the Arabidopsis thaliana lipid droplet oleosin and caleosin proteins AtOle1 and AtClo1. These transformed yeasts overaccumulate lipid droplets, leading to a specific increase in storage lipids. The phenotype of these cells was explored using synchrotron FT-IR microspectroscopy to investigate the dynamics of lipid storage and cellular carbon fluxes reflected as changes in spectral fingerprints. Multivariate statistical analysis of the data showed a clear effect on storage carbohydrates and more specifically, a decrease in glycogen in our modified strains. These observations were confirmed by biochemical quantification of the storage carbohydrates glycogen and trehalose. Our results demonstrate that neutral lipid and storage carbohydrate fluxes are tightly connected and co-regulated. PMID:24040242

  18. Single Cell Synchrotron FT-IR Microspectroscopy Reveals a Link between Neutral Lipid and Storage Carbohydrate Fluxes in S. cerevisiae

    PubMed Central

    Jamme, Frédéric; Vindigni, Jean-David; Méchin, Valérie; Cherifi, Tamazight; Chardot, Thierry; Froissard, Marine

    2013-01-01

    In most organisms, storage lipids are packaged into specialized structures called lipid droplets. These contain a core of neutral lipids surrounded by a monolayer of phospholipids, and various proteins which vary depending on the species. Hydrophobic structural proteins stabilize the interface between the lipid core and aqueous cellular environment (perilipin family of proteins, apolipoproteins, oleosins). We developed a genetic approach using heterologous expression in Saccharomyces cerevisiae of the Arabidopsis thaliana lipid droplet oleosin and caleosin proteins AtOle1 and AtClo1. These transformed yeasts overaccumulate lipid droplets, leading to a specific increase in storage lipids. The phenotype of these cells was explored using synchrotron FT-IR microspectroscopy to investigate the dynamics of lipid storage and cellular carbon fluxes reflected as changes in spectral fingerprints. Multivariate statistical analysis of the data showed a clear effect on storage carbohydrates and more specifically, a decrease in glycogen in our modified strains. These observations were confirmed by biochemical quantification of the storage carbohydrates glycogen and trehalose. Our results demonstrate that neutral lipid and storage carbohydrate fluxes are tightly connected and co-regulated. PMID:24040242

  19. Histone Sprocket Arginine Residues Are Important for Gene Expression, DNA Repair, and Cell Viability in Saccharomyces cerevisiae.

    PubMed

    Hodges, Amelia J; Gallegos, Isaura J; Laughery, Marian F; Meas, Rithy; Tran, Linh; Wyrick, John J

    2015-07-01

    A critical feature of the intermolecular contacts that bind DNA to the histone octamer is the series of histone arginine residues that insert into the DNA minor groove at each superhelical location where the minor groove faces the histone octamer. One of these "sprocket" arginine residues, histone H4 R45, significantly affects chromatin structure in vivo and is lethal when mutated to alanine or cysteine in Saccharomyces cerevisiae (budding yeast). However, the roles of the remaining sprocket arginine residues (H3 R63, H3 R83, H2A R43, H2B R36, H2A R78, H3 R49) in chromatin structure and other cellular processes have not been well characterized. We have genetically characterized mutations in each of these histone residues when introduced either singly or in combination to yeast cells. We find that pairs of arginine residues that bind DNA adjacent to the DNA exit/entry sites in the nucleosome are lethal in yeast when mutated in combination and cause a defect in histone occupancy. Furthermore, mutations in individual residues compromise repair of UV-induced DNA lesions and affect gene expression and cryptic transcription. This study reveals simple rules for how the location and structural mode of DNA binding influence the biological function of each histone sprocket arginine residue. PMID:25971662

  20. The Identification of Transposon-Tagged Mutations in Essential Genes That Affect Cell Morphology in Saccharomyces Cerevisiae

    PubMed Central

    Chun, K. T.; Goebl, M. G.

    1996-01-01

    The yeast Saccharomyces cerevisiae reproduces by budding, and many genes are required for proper bud development. Mutations in some of these genes cause cells to die with an unusual terminal morphology--elongated or otherwise aberrantly shaped buds. To gain insight into bud development, we set out to identify novel genes that encode proteins required for proper bud morphogenesis. Previous studies screened collections of conditional mutations to identify genes required for essential functions, including bud formation. However, genes that are not susceptible to the generation of mutations that cause a conditional phenotype will not be identified in such screens. To identify a more comprehensive collection of mutants, we used transposon mutagenesis to generate a large collection of lethal disruption mutations. This collection was used to identify 209 mutants with disruptions that cause an aberrant terminal bud morphology. The disruption mutations in 33 of these mutants identify three previously uncharacterized genes as essential, and the mutant phenotypes suggest roles for their products in bud morphogenesis. PMID:8770583

  1. High cell density culture with S. cerevisiae CEN.PK113-5D for IL-1β production: optimization, modeling, and physiological aspects.

    PubMed

    Landi, Carmine; Paciello, Lucia; de Alteriis, Elisabetta; Brambilla, Luca; Parascandola, Palma

    2015-02-01

    Saccharomyces cerevisiae CEN.PK113-5D, a strain auxotrophic for uracil belonging to the CEN.PK family of the yeast S. cerevisiae, was cultured in aerated fed-batch reactor as such and once transformed to express human interleukin-1β (IL-1β), aiming at obtaining high cell densities and optimizing IL-1β production. Three different exponentially increasing glucose feeding profiles were tested, all of them "in theory" promoting respiratory metabolism to obtain high biomass/product yield. A non-structured non-segregated model was developed to describe the performance of S. cerevisiae CEN.PK113-5D during the fed-batch process and, in particular, its capability to metabolize simultaneously glucose and ethanol which derived from the precedent batch growth. Our study showed that the proliferative capacity of the yeast population declined along the fed-batch run, as shown by the exponentially decreasing specific growth rates on glucose. Further, a shift towards fermentative metabolism occurred. This shift took place earlier the higher was the feed rate and was more pronounced in the case of the recombinant strain. Determination of some physiological markers (acetate production, intracellular ROS accumulation, catalase activity and cell viability) showed that neither poor oxygenation nor oxidative stress was responsible for the decreased specific growth rate, nor for the shift to fermentative metabolism. PMID:25106469

  2. Cathodic oxygen reduction catalyzed by bacteria in microbial fuel cells.

    PubMed

    Rabaey, Korneel; Read, Suzanne T; Clauwaert, Peter; Freguia, Stefano; Bond, Philip L; Blackall, Linda L; Keller, Jurg

    2008-05-01

    Microbial fuel cells (MFCs) have the potential to combine wastewater treatment efficiency with energetic efficiency. One of the major impediments to MFC implementation is the operation of the cathode compartment, as it employs environmentally unfriendly catalysts such as platinum. As recently shown, bacteria can facilitate sustainable and cost-effective cathode catalysis for nitrate and also oxygen. Here we describe a carbon cathode open to the air, on which attached bacteria catalyzed oxygen reduction. The bacteria present were able to reduce oxygen as the ultimate electron acceptor using electrons provided by the solid-phase cathode. Current densities of up to 2.2 A m(-2) cathode projected surface were obtained (0.303+/-0.017 W m(-2), 15 W m(-3) total reactor volume). The cathodic microbial community was dominated by Sphingobacterium, Acinetobacter and Acidovorax sp., according to 16S rRNA gene clone library analysis. Isolates of Sphingobacterium sp. and Acinetobacter sp. were obtained using H(2)/O(2) mixtures. Some of the pure culture isolates obtained from the cathode showed an increase in the power output of up to three-fold compared to a non-inoculated control, that is, from 0.015+/-0.001 to 0.049+/-0.025 W m(-2) cathode projected surface. The strong decrease in activation losses indicates that bacteria function as true catalysts for oxygen reduction. Owing to the high overpotential for non-catalyzed reduction, oxygen is only to a limited extent competitive toward the electron donor, that is, the cathode. Further research to refine the operational parameters and increase the current density by modifying the electrode surface and elucidating the bacterial metabolism is warranted. PMID:18288216

  3. Metabolism of D-aminoacyl-tRNAs in Escherichia coli and Saccharomyces cerevisiae cells.

    PubMed

    Soutourina, J; Plateau, P; Blanquet, S

    2000-10-20

    In Escherichia coli, tyrosyl-tRNA synthetase is known to esterify tRNA(Tyr) with tyrosine. Resulting d-Tyr-tRNA(Tyr) can be hydrolyzed by a d-Tyr-tRNA(Tyr) deacylase. By monitoring E. coli growth in liquid medium, we systematically searched for other d-amino acids, the toxicity of which might be exacerbated by the inactivation of the gene encoding d-Tyr-tRNA(Tyr) deacylase. In addition to the already documented case of d-tyrosine, positive responses were obtained with d-tryptophan, d-aspartate, d-serine, and d-glutamine. In agreement with this observation, production of d-Asp-tRNA(Asp) and d-Trp-tRNA(Trp) by aspartyl-tRNA synthetase and tryptophanyl-tRNA synthetase, respectively, was established in vitro. Furthermore, the two d-aminoacylated tRNAs behaved as substrates of purified E. coli d-Tyr-tRNA(Tyr) deacylase. These results indicate that an unexpected high number of d-amino acids can impair the bacterium growth through the accumulation of d-aminoacyl-tRNA molecules and that d-Tyr-tRNA(Tyr) deacylase has a specificity broad enough to recycle any of these molecules. The same strategy of screening was applied using Saccharomyces cerevisiae, the tyrosyl-tRNA synthetase of which also produces d-Tyr-tRNA(Tyr), and which, like E. coli, possesses a d-Tyr-tRNA(Tyr) deacylase activity. In this case, inhibition of growth by the various 19 d-amino acids was followed on solid medium. Two isogenic strains containing or not the deacylase were compared. Toxic effects of d-tyrosine and d-leucine were reinforced upon deprivation of the deacylase. This observation suggests that, in yeast, at least two d-amino acids succeed in being transferred onto tRNAs and that, like in E. coli, the resulting two d-aminoacyl-tRNAs are substrates of a same d-aminoacyl-tRNA deacylase. PMID:10918062

  4. Determination of Microbial Growth by Protein Assay in an Air-Cathode Single Chamber Microbial Fuel Cell.

    PubMed

    Li, Na; Kakarla, Ramesh; Moon, Jung Mi; Min, Booki

    2015-07-01

    Microbial fuel cells (MFCs) have gathered attention as a novel bioenergy technology to simultaneously treat wastewater with less sludge production than the conventional activated sludge system. In two different operations of the MFC and aerobic process, microbial growth was determined by the protein assay method and their biomass yields using real wastewater were compared. The biomass yield on the anode electrode of the MFC was 0.02 g-COD-cell/g- COD-substrate and the anolyte planktonic biomass was 0.14 g-COD-cell/g-COD-substrate. An MFC without anode electrode resulted in the biomass yield of 0.07 ± 0.03 g-COD-cell/g-COD-substrate, suggesting that oxygen diffusion from the cathode possibly supported the microbial growth. In a comparative test, the biomass yield under aerobic environment was 0.46 ± 0.07 g-COD-cell/g-COD-substrate, which was about 3 times higher than the total biomass value in the MFC operation. PMID:25674807

  5. In silico model-driven cofactor engineering strategies for improving the overall NADP(H) turnover in microbial cell factories.

    PubMed

    Lakshmanan, Meiyappan; Yu, Kai; Koduru, Lokanand; Lee, Dong-Yup

    2015-10-01

    Optimizing the overall NADPH turnover is one of the key challenges in various value-added biochemical syntheses. In this work, we first analyzed the NADPH regeneration potentials of common cell factories, including Escherichia coli, Saccharomyces cerevisiae, Bacillus subtilis, and Pichia pastoris across multiple environmental conditions and determined E. coli and glycerol as the best microbial chassis and most suitable carbon source, respectively. In addition, we identified optimal cofactor specificity engineering (CSE) enzyme targets, whose cofactors when switched from NAD(H) to NADP(H) improve the overall NADP(H) turnover. Among several enzyme targets, glyceraldehyde-3-phosphate dehydrogenase was recognized as a global candidate since its CSE improved the NADP(H) regeneration under most of the conditions examined. Finally, by analyzing the protein structures of all CSE enzyme targets via homology modeling, we established that the replacement of conserved glutamate or aspartate with serine in the loop region could change the cofactor dependence from NAD(H) to NADP(H). PMID:26254041

  6. Microbial Cryptotopes are Prominent Targets of B-cell Immunity

    PubMed Central

    Rieder, Franz J. J.; Biebl, Julia; Kastner, Marie-Theres; Schneider, Martina; Jungbauer, Christof; Redlberger-Fritz, Monika; Britt, William J.; Kundi, Michael; Steininger, Christoph

    2016-01-01

    B-cell recognition of microbial antigens may be limited by masking of epitopes within three-dimensional structures (cryptotopes). Here we report that unmasking of cryptotopes by unfolding whole cytomegalovirus (CMV) antigen preparations with the chaotropic reagent Urea and probing with immune sera from healthy individuals (n = 109) increased ELISA signals by 36% in comparison to folded CMV antigens (P < 0.001). ELISA signals increased also significantly upon unfolding of S. aureus or E. coli antigens, whereas unfolded influenza H1N1 or respiratory syncitial virus antigens yielded reduced or unchanged reactivity in comparison to folded ones, respectively. Blocking of CMV cryptotope-specific Abs by incubation of an immunoglobuline preparation and three sera with unfolded CMV antigens enhanced clearly the neutralizing capacity of this immunoglobuline preparation against CMV infection. Thus, B-cell immunity frequently targets cryptotopes on CMV but these Abs are non-neutralizing, may reduce the neutralizing effectiveness of pathogen-specific Abs, and increase during immune maturation following primary CMV infection. The observation of functional consequences of Abs specific for cryptotopes may open whole new avenues to a better understanding of the humoral immune response to CMV and development of more effective vaccines and immunoglobuline preparations. PMID:27539094

  7. Microbial Cryptotopes are Prominent Targets of B-cell Immunity.

    PubMed

    Rieder, Franz J J; Biebl, Julia; Kastner, Marie-Theres; Schneider, Martina; Jungbauer, Christof; Redlberger-Fritz, Monika; Britt, William J; Kundi, Michael; Steininger, Christoph

    2016-01-01

    B-cell recognition of microbial antigens may be limited by masking of epitopes within three-dimensional structures (cryptotopes). Here we report that unmasking of cryptotopes by unfolding whole cytomegalovirus (CMV) antigen preparations with the chaotropic reagent Urea and probing with immune sera from healthy individuals (n = 109) increased ELISA signals by 36% in comparison to folded CMV antigens (P < 0.001). ELISA signals increased also significantly upon unfolding of S. aureus or E. coli antigens, whereas unfolded influenza H1N1 or respiratory syncitial virus antigens yielded reduced or unchanged reactivity in comparison to folded ones, respectively. Blocking of CMV cryptotope-specific Abs by incubation of an immunoglobuline preparation and three sera with unfolded CMV antigens enhanced clearly the neutralizing capacity of this immunoglobuline preparation against CMV infection. Thus, B-cell immunity frequently targets cryptotopes on CMV but these Abs are non-neutralizing, may reduce the neutralizing effectiveness of pathogen-specific Abs, and increase during immune maturation following primary CMV infection. The observation of functional consequences of Abs specific for cryptotopes may open whole new avenues to a better understanding of the humoral immune response to CMV and development of more effective vaccines and immunoglobuline preparations. PMID:27539094

  8. Compost in plant microbial fuel cell for bioelectricity generation.

    PubMed

    Moqsud, M A; Yoshitake, J; Bushra, Q S; Hyodo, M; Omine, K; Strik, David

    2015-02-01

    Recycling of organic waste is an important topic in developing countries as well as developed countries. Compost from organic waste has been used for soil conditioner. In this study, an experiment has been carried out to produce green energy (bioelectricity) by using paddy plant microbial fuel cells (PMFCs) in soil mixed with compost. A total of six buckets filled with the same soil were used with carbon fiber as the electrodes for the test. Rice plants were planted in five of the buckets, with the sixth bucket containing only soil and an external resistance of 100 ohm was used for all cases. It was observed that the cells with rice plants and compost showed higher values of voltage and power density with time. The highest value of voltage showed around 700 mV when a rice plant with 1% compost mixed soil was used, however it was more than 95% less in the case of no rice plant and without compost. Comparing cases with and without compost but with the same number of rice plants, cases with compost depicted higher voltage to as much as 2 times. The power density was also 3 times higher when the compost was used in the paddy PMFCs which indicated the influence of compost on bio-electricity generation. PMID:25443096

  9. Microbial fuel cells with highly active aerobic biocathodes

    NASA Astrophysics Data System (ADS)

    Milner, Edward M.; Popescu, Dorin; Curtis, Tom; Head, Ian M.; Scott, Keith; Yu, Eileen H.

    2016-08-01

    Microbial fuel cells (MFCs), which convert organic waste to electricity, could be used to make the wastewater infrastructure more energy efficient and sustainable. However, platinum and other non-platinum chemical catalysts used for the oxygen reduction reaction (ORR) at the cathode of MFCs are unsustainable due to their high cost and long-term degradation. Aerobic biocathodes, which use microorganisms as the biocatalysts for cathode ORR, are a good alternative to chemical catalysts. In the current work, high-performing aerobic biocathodes with an onset potential for the ORR of +0.4 V vs. Ag/AgCl were enriched from activated sludge in electrochemical half-cells poised at -0.1 and + 0.2 V vs. Ag/AgCl. Gammaproteobacteria, distantly related to any known cultivated gammaproteobacterial lineage, were identified as dominant in these working electrode biofilms (23.3-44.3% of reads in 16S rRNA gene Ion Torrent libraries), and were in very low abundance in non-polarised control working electrode biofilms (0.5-0.7%). These Gammaproteobacteria were therefore most likely responsible for the high activity of biologically catalysed ORR. In MFC tests, a high-performing aerobic biocathode increased peak power 9-fold from 7 to 62 μW cm-2 in comparison to an unmodified carbon cathode, which was similar to peak power with a platinum-doped cathode at 70 μW cm-2.

  10. Increased electrical output when a bacterial ABTS oxidizer is used in a microbial fuel cell

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Microbial fuel cells (MFCs) are a technology that provides electrical energy from the microbial oxidation of organic compounds. Most MFCs use oxygen as the oxidant in the cathode chamber. The present study examined the formation in culture of an unidentified bacterial oxidant and investigated the ...

  11. Mechanisms of Cell Cycle Control Revealed by a Systematic and Quantitative Overexpression Screen in S. cerevisiae

    PubMed Central

    Niu, Wei; Li, Zhihua; Zhan, Wenjing; Iyer, Vishwanath R.; Marcotte, Edward M.

    2008-01-01

    Regulation of cell cycle progression is fundamental to cell health and reproduction, and failures in this process are associated with many human diseases. Much of our knowledge of cell cycle regulators derives from loss-of-function studies. To reveal new cell cycle regulatory genes that are difficult to identify in loss-of-function studies, we performed a near-genome-wide flow cytometry assay of yeast gene overexpression-induced cell cycle delay phenotypes. We identified 108 genes whose overexpression significantly delayed the progression of the yeast cell cycle at a specific stage. Many of the genes are newly implicated in cell cycle progression, for example SKO1, RFA1, and YPR015C. The overexpression of RFA1 or YPR015C delayed the cell cycle at G2/M phases by disrupting spindle attachment to chromosomes and activating the DNA damage checkpoint, respectively. In contrast, overexpression of the transcription factor SKO1 arrests cells at G1 phase by activating the pheromone response pathway, revealing new cross-talk between osmotic sensing and mating. More generally, 92%–94% of the genes exhibit distinct phenotypes when overexpressed as compared to their corresponding deletion mutants, supporting the notion that many genes may gain functions upon overexpression. This work thus implicates new genes in cell cycle progression, complements previous screens, and lays the foundation for future experiments to define more precisely roles for these genes in cell cycle progression. PMID:18617996

  12. Anhydrobiosis in yeast: cell wall mannoproteins are important for yeast Saccharomyces cerevisiae resistance to dehydration.

    PubMed

    Borovikova, Diana; Teparić, Renata; Mrša, Vladimir; Rapoport, Alexander

    2016-08-01

    The state of anhydrobiosis is linked with the reversible delay of metabolism as a result of strong dehydration of cells, and is widely distributed in nature. A number of factors responsible for the maintenance of organisms' viability in these conditions have been revealed. This study was directed to understanding how changes in cell wall structure may influence the resistance of yeasts to dehydration-rehydration. Mutants lacking various cell wall mannoproteins were tested to address this issue. It was revealed that mutants lacking proteins belonging to two structurally and functionally unrelated groups (proteins non-covalently attached to the cell wall, and Pir proteins) possessed significantly lower cell resistance to dehydration-rehydration than the mother wild-type strain. At the same time, the absence of the GPI-anchored cell wall protein Ccw12 unexpectedly resulted in an increase of cell resistance to this treatment; this phenomenon is explained by the compensatory synthesis of chitin. The results clearly indicate that the cell wall structure/composition relates to parameters strongly influencing yeast viability during the processes of dehydration-rehydration, and that damage to cell wall proteins during yeast desiccation can be an important factor leading to cell death. Copyright © 2016 John Wiley & Sons, Ltd. PMID:27510749

  13. Cadmium-induced cell killing in Sacharomyces cerevisiae involves increases in intracellular NO levels.

    PubMed

    Wu, Lihua; Chen, Yanfei; Gao, Huixian; Yin, Jingjing; Huang, Liqun

    2016-03-01

    Cadmium is a widespread environmental pollutant and poses some potential risks to human health. However, the signaling events controlling cadmium toxicity are not fully understood. In this study, we examined the effect of cadmium chloride on cell viability and the intracellular nitric oxide (NO) level in yeast cells. The results showed that exposure of yeast cells to cadmium (0-100 μM) could induce cell killing with significantly increased intracellular NO levels. Morphological analysis of the nuclei with 4('),6-diamidino-2-phenylindole staining and DNA strand breaks analysis showed that cadmium at 50 μM can induce cell apoptosis in yeast cells. Treatment of yeast cells with cadmium (50 μM) and the nitric oxide scavenger c-PTIO [2-(4-carboxyphenyl)-4,4,5,5-teramethylimidazoline-1-oxyl-3-oxide; 0.2 mM] showed that c-PTIO attenuated the cadmium-induced cell killing. Our findings indicated that cadmium-induced yeast cell killing is mediated by a directly increased intracellular NO level. PMID:26872495

  14. Cytoplasmic proteasomes are not indispensable for cell growth in Saccharomyces cerevisiae

    SciTech Connect

    Tsuchiya, Hikaru; Arai, Naoko; Tanaka, Keiji Saeki, Yasushi

    2013-07-05

    Highlights: •We succeeded to control the proteasome localization by the anchor-away technique. •Nuclear proteasome-depleted cells showed a lethal phenotype. •Cytoplasmic proteasomes are not indispensable for cell growth in dividing cells. -- Abstract: The 26S proteasome is an essential protease complex responsible for the degradation of ubiquitinated proteins in eukaryotic cells. In rapidly proliferating yeast cells, proteasomes are mainly localized in the nucleus, but the biological significance of the proteasome localization is still unclear. In this study, we investigated the relationship between the proteasome localization and the functions by the anchor-away technique, a ligand-dependent sequestration of a target protein into specific compartment(s). Anchoring of the proteasome to the plasma membrane or the ribosome resulted in conditional depletion of the nuclear proteasomes, whereas anchoring to histone resulted in the proteasome sequestration into the nucleus. We observed that the accumulation of ubiquitinated proteins in all the proteasome-targeted cells, suggesting that both the nuclear and cytoplasmic proteasomes have proteolytic functions and that the ubiquitinated proteins are produced and degraded in each compartment. Consistent with previous studies, the nuclear proteasome-depleted cells exhibited a lethal phenotype. In contrast, the nuclear sequestration of the proteasome resulted only in a mild growth defect, suggesting that the cytoplasmic proteasomes are not basically indispensable for cell growth in rapidly growing yeast cells.

  15. Large Scale Identification of Genes Involved in Cell Surface Biosynthesis and Architecture in Saccharomyces Cerevisiae

    PubMed Central

    Lussier, M.; White, A. M.; Sheraton, J.; di-Paolo, T.; Treadwell, J.; Southard, S. B.; Horenstein, C. I.; Chen-Weiner, J.; Ram, AFJ.; Kapteyn, J. C.; Roemer, T. W.; Vo, D. H.; Bondoc, D. C.; Hall, J.; Wei Zhong, W.; Sdicu, A. M.; Davies, J.; Klis, F. M.; Robbins, P. W.; Bussey, H.

    1997-01-01

    The sequenced yeast genome offers a unique resource for the analysis of eukaryotic cell function and enables genome-wide screens for genes involved in cellular processes. We have identified genes involved in cell surface assembly by screening transposon-mutagenized cells for altered sensitivity to calcofluor white, followed by supplementary screens to further characterize mutant phenotypes. The mutated genes were directly retrieved from genomic DNA and then matched uniquely to a gene in the yeast genome database. Eighty-two genes with apparent perturbation of the cell surface were identified, with mutations in 65 of them displaying at least one further cell surface phenotype in addition to their modified sensitivity to calcofluor. Fifty of these genes were previously known, 17 encoded proteins whose function could be anticipated through sequence homology or previously recognized phenotypes and 15 genes had no previously known phenotype. PMID:9335584

  16. Microbial desalination cells for improved performance in wastewater treatment, electricity production, and desalination.

    PubMed

    Luo, Haiping; Xu, Pei; Roane, Timberley M; Jenkins, Peter E; Ren, Zhiyong

    2012-02-01

    The low conductivity and alkalinity in municipal wastewater significantly limit power production from microbial fuel cells (MFCs). This study integrated desalination with wastewater treatment and electricity production in a microbial desalination cell (MDC) by utilizing the mutual benefits among the above functions. When using wastewater as the sole substrate, the power output from the MDC (8.01 W/m(3)) was four times higher than a control MFC without desalination function. In addition, the MDC removed 66% of the salts and improved COD removal by 52% and Coulombic efficiency by 131%. Desalination in MDCs improved wastewater characteristics by increasing the conductivity by 2.5 times and stabilizing anolyte pH, which therefore reduced system resistance and maintained microbial activity. Microbial community analysis revealed a more diverse anode microbial structure in the MDC than in the MFC. The results demonstrated that MDC can serve as a viable option for integrated wastewater treatment, energy production, and desalination. PMID:22178493

  17. Is resistance futile? Changing external resistance does not improve microbial fuel cell performance.

    PubMed

    Lyon, Delina Y; Buret, Francois; Vogel, Timothy M; Monier, Jean-Michel

    2010-04-01

    Microbial fuel cells (MFCs) show promise as an alternative to conventional batteries for point source electricity generation. A better understanding of the relationship between the microbiological and electrical aspects of fuels cells is needed prior to successful MFC application. Here, we observed the effects of external resistance on power production and the anodic biofilm community structure. Large differences in the external resistance affected both power production and microbial community structure. After the establishment of the anodic microbial community, change in external resistance (from low to high and vice versa) changed the anodic microbial community structure, but the resulting community did not resemble the communities established at that same external resistance. Different microbial community structures, established under different external resistances, resulted in similar power production, demonstrating the flexibility of the MFC system. PMID:19783225

  18. Real-time viable-cell mass monitoring in high-cell-density fed-batch glutathione fermentation by Saccharomyces cerevisiae T65 in industrial complex medium.

    PubMed

    Xiong, Zhi-Qiang; Guo, Mei-Jin; Guo, Yuan-Xin; Chu, Ju; Zhuang, Ying-Ping; Zhang, Si-Liang

    2008-04-01

    An on-line monitoring of viable-cell mass in high-cell-density fed-batch cultivations of Saccharomyces cerevisiae grown on an industrial complex medium was performed with an in situ capacitance probe fitted to a 50-l fermentor. Conventional off-line biomass determinations of several parameters, including dry cell weight (DCW), optical density at 600 nm wavelength (OD(600)), packed mycelial volume (PMV) and number of colony forming units (CFU), were performed throughout the bioprocess and then compared with on-line viable-cell concentrations measured using a capacitance probe. Capacitance versus viable biomass and all off-line biomass assay values were compared during glutathione fermentation in industrial complex culture media. As a result, the relationship between the number of colony forming units and capacitance with a correlation coefficient (R) of 0.995 was achieved. Simultaneously, compared with those determined by at-line indirect estimation methods including oxygen uptake rate (OUR) and carbon dioxide evolution rate (CER), the specific growth rates estimated by on-line capacitance measurement could be more reliable during glutathione fermentation. Therefore, it is concluded that a capacitance probe is a practical tool for real-time viable biomass monitoring in high-cell-density fed-batch cultivation in a complex medium. PMID:18499059

  19. Increased power density from a spiral wound microbial fuel cell.

    PubMed

    Jia, Boyang; Hu, Dawei; Xie, Beizhen; Dong, Kun; Liu, Hong

    2013-03-15

    Using Microbial fuel cell (MFC) to convert organic and inorganic matter into electricity is of great interest for powering portable devices, which is now still limited by the output of MFC. In this study, a spiral wound MFC (SWMFC) with relatively large volume normalized surface area of separator (4.2 cm(2)/ml) was fabricated to enhance power generation. Compared with double-membrane MFC (DMMFC) and conventional double chamber MFC (DCMFC), the power density of SWMFC increased by 42% and 99% resulted from its lower internal resistance. Besides larger separator area, the better performance of SWMFC benefited from its structure sandwiching the cathodes between two separators. This point was proved again by a comparison of another DCMFC and a triple chamber MFC (TCMFC) as well as a simulation using finite element method. Moreover, the feature of SWMFC was more convenient and compact to scale up. Therefore, SWMFC provides a promising configuration for high power output as a portable power source. PMID:23116542

  20. Polymer coatings as separator layers for microbial fuel cell cathodes

    NASA Astrophysics Data System (ADS)

    Watson, Valerie J.; Saito, Tomonori; Hickner, Michael A.; Logan, Bruce E.

    2011-03-01

    Membrane separators reduce oxygen flux from the cathode into the anolyte in microbial fuel cells (MFCs), but water accumulation and pH gradients between the separator and cathode reduces performance. Air cathodes were spray-coated (water-facing side) with anion exchange, cation exchange, and neutral polymer coatings of different thicknesses to incorporate the separator into the cathode. The anion exchange polymer coating resulted in greater power density (1167 ± 135 mW m-2) than a cation exchange coating (439 ± 2 mW m-2). This power output was similar to that produced by a Nafion-coated cathode (1114 ± 174 mW m-2), and slightly lower than the uncoated cathode (1384 ± 82 mW m-2). Thicker coatings reduced oxygen diffusion into the electrolyte and increased coulombic efficiency (CE = 56-64%) relative to an uncoated cathode (29 ± 8%), but decreased power production (255-574 mW m-2). Electrochemical characterization of the cathodes ex situ to the MFC showed that the cathodes with the lowest charge transfer resistance and the highest oxygen reduction activity produced the most power in MFC tests. The results on hydrophilic cathode separator layers revealed a trade off between power and CE. Cathodes coated with a thin coating of anion exchange polymer show promise for controlling oxygen transfer while minimally affecting power production.

  1. Carbon Material Optimized Biocathode for Improving Microbial Fuel Cell Performance

    PubMed Central

    Tursun, Hairti; Liu, Rui; Li, Jing; Abro, Rashid; Wang, Xiaohui; Gao, Yanmei; Li, Yuan

    2016-01-01

    To improve the performance of microbial fuel cells (MFCs), the biocathode electrode material of double-chamber was optimized. Alongside the basic carbon fiber brush, three carbon materials namely graphite granules, activated carbon granules (ACG) and activated carbon powder, were added to the cathode-chambers to improve power generation. The result shows that the addition of carbon materials increased the amount of available electroactive microbes on the electrode surface and thus promote oxygen reduction rate, which improved the generation performance of the MFCs. The Output current (external resistance = 1000 Ω) greatly increased after addition of the three carbon materials and maximum power densities in current stable phase increased by 47.4, 166.1, and 33.5%, respectively. Additionally, coulombic efficiencies of the MFC increased by 16.3, 64.3, and 20.1%, respectively. These results show that MFC when optimized with ACG show better power generation, higher chemical oxygen demands removal rate and coulombic efficiency. PMID:26858695

  2. Carbon fiber enhanced bioelectricity generation in soil microbial fuel cells.

    PubMed

    Li, Xiaojing; Wang, Xin; Zhao, Qian; Wan, Lili; Li, Yongtao; Zhou, Qixing

    2016-11-15

    The soil microbial fuel cell (MFC) is a promising biotechnology for the bioelectricity recovery as well as the remediation of organics contaminated soil. However, the electricity production and the remediation efficiency of soil MFC are seriously limited by the tremendous internal resistance of soil. Conductive carbon fiber was mixed with petroleum hydrocarbons contaminated soil and significantly enhanced the performance of soil MFC. The maximum current density, the maximum power density and the accumulated charge output of MFC mixed carbon fiber (MC) were 10, 22 and 16 times as high as those of closed circuit control due to the carbon fiber productively assisted the anode to collect the electron. The internal resistance of MC reduced by 58%, 83% of which owed to the charge transfer resistance, resulting in a high efficiency of electron transfer from soil to anode. The degradation rates of total petroleum hydrocarbons enhanced by 100% and 329% compared to closed and opened circuit controls without the carbon fiber respectively. The effective range of remediation and the bioelectricity recovery was extended from 6 to 20cm with the same area of air-cathode. The mixed carbon fiber apparently enhanced the bioelectricity generation and the remediation efficiency of soil MFC by means of promoting the electron transfer rate from soil to anode. The use of conductively functional materials (e.g. carbon fiber) is very meaningful for the remediation and bioelectricity recovery in the bioelectrochemical remediation. PMID:27162144

  3. Cadmium (II) removal mechanisms in microbial electrolysis cells.

    PubMed

    Colantonio, Natalie; Kim, Younggy

    2016-07-01

    Cadmium is a toxic heavy metal, causing serious environmental and human health problems. Conventional methods for removing cadmium from wastewater are expensive and inefficient for low concentrations. Microbial electrolysis cells (MECs) can simultaneously treat wastewater, produce hydrogen gas, and remove heavy metals with low energy requirements. Lab-scale MECs were operated to remove cadmium under various electric conditions: applied voltages of 0.4, 0.6, 0.8, and 1.0 V; and a fixed cathode potential of -1.0 V vs. Ag/AgCl. Regardless of the electric condition, rapid removal of cadmium was demonstrated (50-67% in 24 h); however, cadmium concentration in solution increased after the electric current dropped with depleted organic substrate under applied voltage conditions. For the fixed cathode potential, the electric current was maintained even after substrate depletion and thus cadmium concentration did not increase. These results can be explained by three different removal mechanisms: cathodic reduction; Cd(OH)2 precipitation; and CdCO3 precipitation. When the current decreased with depleted substrates, local pH at the cathode was no longer high due to slowed hydrogen evolution reaction (2H(+)+2e(-)→H2); thus, the precipitated Cd(OH)2 and CdCO3 started dissolving. To prevent their dissolution, sufficient organic substrates should be provided when MECs are used for cadmium removal. PMID:26970043

  4. Microbial fuel cell energy harvesting using synchronous flyback converter

    NASA Astrophysics Data System (ADS)

    Alaraj, Muhannad; Ren, Zhiyong Jason; Park, Jae-Do

    2014-02-01

    Microbial Fuel Cells (MFCs) use biodegradable substrates, such as wastewater and marine sediments to generate electrical energy. To harvest more energy from an MFC, power electronic converters have recently been used to replace resistors or charge pumps, because they have superior controllability on MFC's operating point and higher efficiency in energy storage for different applications. Conventional diode-based energy harvesters suffer from low efficiency because of the energy losses through the diode. Replacing the diode with a MOSFET can reduce the conduction loss, but it requires an isolated gate signal to control the floating secondary MOSFET, which makes the control circuitry complex. This study presents a new MFC energy harvesting regime using a synchronous flyback converter, which implements a transformer-based harvester with much simpler configuration and improves harvesting efficiency by 37.6% compared to a diode based boost converter, from 33.5% to 46.1%. The proposed harvester was able to store 2.27 J in the output capacitor out of 4.91 J generated energy from the MFC, while the boost converter can capture 1.67 J from 4.95 J.

  5. Shift of voltage reversal in stacked microbial fuel cells

    NASA Astrophysics Data System (ADS)

    An, Junyeong; Kim, Bongkyu; Chang, In Seop; Lee, Hyung-Sool

    2015-03-01

    We proved that sluggish kinetics on the cathode and the imbalance of cathode kinetics cause voltage reversal in a stacked microbial fuel cell (MFC) equipped with a non-Pt cathode. Catholyte aeration to a unit MFC against passive air diffusion to the cathode in the other unit MFC shifted voltage reversal between the two units, due to improved mass transport and O2 concentration effects in the aerated MFC. The shifted voltage reversal returned to an original status when catholyte aeration was stopped. A Pt-coated cathode increased the rate of oxygen reduction reaction (ORR) by a factor of ∼20, as compared to the non-Pt cathode. As a result, the anodic reaction rate that became slower than the rate on the Pt-cathode limited current density to overpotential in the stacked MFC equipped with the Pt-cathode. This work shows that dominant kinetic bottlenecks, which are the primary cause of voltage reversal, can be shifted between individual MFCs of stacked MFCs or electrodes depending on relative kinetics.

  6. Tubular bamboo charcoal for anode in microbial fuel cells

    NASA Astrophysics Data System (ADS)

    Zhang, Jun; Li, Jun; Ye, Dingding; Zhu, Xun; Liao, Qiang; Zhang, Biao

    2014-12-01

    The anode material plays a significant role in determining the performance of microbial fuel cells (MFCs). In this study, the bamboo charcoal tube is proposed as a novel anode substrate by carbonizing the natural bamboo. Its surface functional groups, biocompatibility and internal resistance are thoroughly investigated. Performance of the MFCs with a conventional graphite tube anode and a bamboo charcoal tube anode is also compared. The results indicate that the tubular bamboo charcoal anode exhibits advantages over the graphite tube anode in terms of rougher surface, superior biocompatibility and smaller total internal resistance. Moreover, the X-ray photoelectron spectroscopy (XPS) analysis for the bamboo charcoal reveals that the introduced C-N bonds facilitate the electron transfer between the biofilm and electrodes. As a result, the MFC with a bamboo charcoal tube anode achieves a 50% improvement in the maximum power density over the graphite tube case. Furthermore, scale-up of the bamboo charcoal tube anode is demonstrated by employing a bundle of tubular bamboo charcoal to reach higher power output.

  7. Carbon Material Optimized Biocathode for Improving Microbial Fuel Cell Performance.

    PubMed

    Tursun, Hairti; Liu, Rui; Li, Jing; Abro, Rashid; Wang, Xiaohui; Gao, Yanmei; Li, Yuan

    2016-01-01

    To improve the performance of microbial fuel cells (MFCs), the biocathode electrode material of double-chamber was optimized. Alongside the basic carbon fiber brush, three carbon materials namely graphite granules, activated carbon granules (ACG) and activated carbon powder, were added to the cathode-chambers to improve power generation. The result shows that the addition of carbon materials increased the amount of available electroactive microbes on the electrode surface and thus promote oxygen reduction rate, which improved the generation performance of the MFCs. The Output current (external resistance = 1000 Ω) greatly increased after addition of the three carbon materials and maximum power densities in current stable phase increased by 47.4, 166.1, and 33.5%, respectively. Additionally, coulombic efficiencies of the MFC increased by 16.3, 64.3, and 20.1%, respectively. These results show that MFC when optimized with ACG show better power generation, higher chemical oxygen demands removal rate and coulombic efficiency. PMID:26858695

  8. Innovative microbial fuel cell for energy harvesting and corrosion protection

    NASA Astrophysics Data System (ADS)

    Kung, Chih-Chien; Liu, Chung-Chiun; Yu, Xiong

    2011-06-01

    Microbial Fuel cells (MFCs) are batteries driven by bacteria. MFCs have the potential of powering small sensors in remote areas and disposing organic waste safely by harvesting the energy stored in the waste products. From previous research in this field, a few important factors for MFC performance have been identified. These include the internal resistance of MFC, the surface area of anode with catalyst for the biofilm development, the type and number of bacteria, and the abundance of nutritional supplies to the bacteria. This paper describes the design of a novel single chamber MFC (SMFC) with carbon electrodes. Experiments were conducted to establish the relationship between each parameter and the power production. It is shown here that this SCMFC can generate electrical current without the use of PEM membranes or additives; the maximum voltage of around 411 mV can be achieved at the room temperature. These results also measured a various parameters such as pH, dissolved oxygen and solution conductivity during the operation of SMFC. Finally, experiment was conducted to evaluate an innovative concept of using MFC for corrosion protection.

  9. Voltage balancing strategies for serial connection of microbial fuel cells

    NASA Astrophysics Data System (ADS)

    Khaled, Firas; Ondel, Olivier; Allard, Bruno; Buret, François

    2015-07-01

    The microbial fuel cell (MFC) converts electrochemically organic matter into electricity by means of metabolisms of bacteria. The MFC power output is limited by low voltage and low current characteristics in the range of microwatts or milliwatts per litre. In order to produce a sufficient voltage level (>1.5 V) and sufficient power to supply real applications such as autonomous sensors, it is necessary to either scale-up one single unit or to connect multiple units together. Many topologies of connection are possible as the serial association to improve the output voltage, or the parallel connection to improve the output current or the series/parallel connection to step-up both voltage and current. The association of MFCs in series is a solution to increase the voltage to an acceptable value and to mutualize the unit's output power. The serial association of a large number of MFCs presents several issues. The first one is the hydraulic coupling among MFCs when they share the same substrate. The second one is the dispersion between generators that lead to a non-optimal stack efficiency because the maximum power point (MPP) operation of all MFCs is not permitted. Voltage balancing is a solution to compensate non-uniformities towards MPP. This paper presents solutions to improve the efficiency of a stack of serially connected MFCs through a voltage-balancing circuit. Contribution to the topical issue "Electrical Engineering Symposium (SGE 2014)", edited by Adel Razek

  10. Electricity generation from rapeseed straw hydrolysates using microbial fuel cells.

    PubMed

    Jablonska, Milena A; Rybarczyk, Maria K; Lieder, Marek

    2016-05-01

    Rapeseed straw is an attractive fuel material for microbial fuel cells (MFCs) due to its high content of carbohydrates (more than 60% carbohydrates). This study has demonstrated that reducing sugars can be efficiently extracted from raw rapeseed straw by combination of hydrothermal pretreatment and enzymatic hydrolysis followed by utilization as a fuel in two-chamber MFCs for electrical power generation. The most efficient method of saccharification of this lignocellulosic biomass (17%) turned out hydrothermal pretreatment followed by enzymatic hydrolysis. Electricity was produced using hydrolysate concentrations up to 150mg/dm(3). The power density reached 54mW/m(2), while CEs ranged from 60% to 10%, corresponding to the initial reducing sugar concentrations of 10-150mg/dm(3). The COD degradation rates based on charge calculation increased from 0.445gCOD/m(2)/d for the hydrolysate obtained with the microwave treatment to 0.602gCOD/m(2)/d for the most efficient combination of hydrothermal treatment followed by enzymatic hydrolysis. PMID:26930033

  11. Submersible microbial fuel cell for electricity production from sewage sludge.

    PubMed

    Zhang, Yifeng; Olias, Lola Gonzalez; Kongjan, Prawit; Angelidaki, Irini

    2011-01-01

    A submersible microbial fuel cell (SMFC) was utilized to treat sewage sludge and simultaneously generate electricity. Stable power generation (145 +/- 5 mW/m2, 470 omega) was produced continuously from raw sewage sludge for 5.5 days. The maximum power density reached 190 +/- 5 mW/m2. The corresponding total chemical oxygen demand (TCOD) removal efficiency was 78.1 +/- 0.2% with initial TCOD of 49.7 g/L. The power generation of SMFC was depended on the sludge concentration, while dilution of the raw sludge resulted in higher power density. The maximum power density was saturated at sludge concentration of 17 g-TCOD/L, where 290 mw/m2 was achieved. When effluents from an anaerobic digester that was fed with raw sludge were used as substrate in the SMFC, a maximum power density of 318 mW/m2, and a final TCOD removal of 71.9 +/- 0.2% were achieved. These results have practical implications for development of an effective system to treat sewage sludge and simultaneously recover energy. PMID:22053457

  12. Investigating Microbial Fuel Cell Bioanode Performance Under Different Cathode Conditions

    SciTech Connect

    Borole, Abhijeet P; Hamilton, Choo Yieng; Aaron, D; Tsouris, Costas

    2009-01-01

    A compact, three-in-one, flow-through, porous, electrode design with minimal electrode spacing and minimal dead volume was implemented to develop a microbial fuel cell (MFC) with improved anode performance. A biofilm-dominated anode consortium enriched under a multimode, continuous-flow regime was used. The increase in the power density of the MFC was investigated by changing the cathode (type, as well as catholyte strength) to determine whether anode was limiting. The power density obtained with an air-breathing cathode was 56 W/m3 of net anode volume (590 mW/m2) and 203 W/m3 (2160 mW/m2) with a 50-mM ferricyanide- based cathode. Increasing the ferricyanide concentration and ionic strength further increased the power density, reaching 304 W/m3 (3220 mW/m2, with 200 mM ferricyanide and 200 mM buffer concentration). The increasing trend in the power density indicated that the anode was not limiting and that higher power densities could be obtained using cathodes capable of higher rates of oxidation. The internal solution resistance for the MFC was 5 6 X, which supported the improved performance of the anode design. A new parameter defined as the ratio of projected surface area to total anode volume is suggested as a design parameter to relate volumetric and area-based power densities and to enable comparison of various MFC configurations.

  13. Influence of anode surface chemistry on microbial fuel cell operation.

    PubMed

    Santoro, Carlo; Babanova, Sofia; Artyushkova, Kateryna; Cornejo, Jose A; Ista, Linnea; Bretschger, Orianna; Marsili, Enrico; Atanassov, Plamen; Schuler, Andrew J

    2015-12-01

    Self-assembled monolayers (SAMs) modified gold anodes are used in single chamber microbial fuel cells for organic removal and electricity generation. Hydrophilic (N(CH3)3(+), OH, COOH) and hydrophobic (CH3) SAMs are examined for their effect on bacterial attachment, current and power output. The different substratum chemistry affects the community composition of the electrochemically active biofilm formed and thus the current and power output. Of the four SAM-modified anodes tested, N(CH3)3(+) results in the shortest start up time (15 days), highest current achieved (225 μA cm(-2)) and highest MFC power density (40 μW cm(-2)), followed by COOH (150 μA cm(-2) and 37 μW cm(-2)) and OH (83 μA cm(-2) and 27 μW cm(-2)) SAMs. Hydrophobic SAM decreases electrochemically active bacteria attachment and anode performance in comparison to hydrophilic SAMs (CH3 modified anodes 7 μA cm(-2) anodic current and 1.2 μW cm(-2) MFC's power density). A consortium of Clostridia and δ-Proteobacteria is found on all the anode surfaces, suggesting a synergistic cooperation under anodic conditions. PMID:26025340

  14. Impact of volatile fatty acids on microbial electrolysis cell performance.

    PubMed

    Yang, Nan; Hafez, Hisham; Nakhla, George

    2015-10-01

    This study investigated the performance of microbial electrolysis cells (MECs) fed with three common fermentation products: acetate, butyrate, and propionate. Each substrate was fed to the reactor for three consecutive-batch cycles. The results showed high current densities for acetate, but low current densities for butyrate and propionate (maximum values were 6.0 ± 0.28, 2.5 ± 0.06, 1.6 ± 0.14 A/m(2), respectively). Acetate also showed a higher coulombic efficiency of 87 ± 5.7% compared to 72 ± 2.0 and 51 ± 6.4% for butyrate and propionate, respectively. This paper also revealed that acetate could be easily oxidized by anode respiring bacteria in MEC, while butyrate and propionate could not be oxidized to the same degree. The utilization rate of the substrates in MEC followed the order: acetate > butyrate > propionate. The ratio of suspended biomass to attached biomass was approximately 1:4 for all the three substrates. PMID:26159302

  15. Scale-up of phosphate remobilization from sewage sludge in a microbial fuel cell.

    PubMed

    Happe, Manuel; Sugnaux, Marc; Cachelin, Christian Pierre; Stauffer, Marc; Zufferey, Géraldine; Kahoun, Thomas; Salamin, Paul-André; Egli, Thomas; Comninellis, Christos; Grogg, Alain-François; Fischer, Fabian

    2016-01-01

    Phosphate remobilization from digested sewage sludge containing iron phosphate was scaled-up in a microbial fuel cell (MFC). A 3litre triple chambered MFC was constructed. This reactor was operated as a microbial fuel cell and later as a microbial electrolysis cell to accelerate cathodic phosphate remobilization. Applying an additional voltage and exceeding native MFC power accelerated chemical base formation and the related phosphate remobilization rate. The electrolysis approach was extended using a platinum-RVC cathode. The pH rose to 12.6 and phosphate was recovered by 67% in 26h. This was significantly faster than using microbial fuel cell conditions. Shrinking core modelling particle fluid kinetics showed that the reaction resistance has to move inside the sewage sludge particle for considerable rate enhancement. Remobilized phosphate was subsequently precipitated as struvite and inductively coupled plasma mass spectrometry indicated low levels of cadmium, lead, and other metals as required by law for recycling fertilizers. PMID:26519694

  16. Saccharomyces cerevisiae MPT5 and SSD1 function in parallel pathways to promote cell wall integrity.

    PubMed Central

    Kaeberlein, Matt; Guarente, Leonard

    2002-01-01

    Yeast MPT5 (UTH4) is a limiting component for longevity. We show here that MPT5 also functions to promote cell wall integrity. Loss of Mpt5p results in phenotypes associated with a weakened cell wall, including sorbitol-remedial temperature sensitivity and sensitivities to calcofluor white and sodium dodecyl sulfate. Additionally, we find that mutation of MPT5, in the absence of SSD1-V, is lethal in combination with loss of either Ccr4p or Swi4p. These synthetic lethal interactions are suppressed by the SSD1-V allele. Furthermore, we have provided evidence that the short life span caused by loss of Mpt5p is due to a weakened cell wall. This cell wall defect may be the result of abnormal chitin biosynthesis or accumulation. These analyses have defined three genetic pathways that function in parallel to promote cell integrity: an Mpt5p-containing pathway, an Ssd1p-containing pathway, and a Pkc1p-dependent pathway. This work also provides evidence that post-transcriptional regulation is likely to be important both for maintaining cell integrity and for promoting longevity. PMID:11805047

  17. Enhanced power production from microbial fuel cells with high cell density culture.

    PubMed

    Zhai, Dan-Dan; Li, Bing; Sun, Jian-Zhong; Sun, De-Zhen; Si, Rong-Wei; Yong, Yang-Chun

    2016-01-01

    Improvement of power production in a microbial fuel cell (MFC) with a high cell density culture strategy was developed. By using high cell density culture, the voltage output and power density output of the MFC were enhanced about 0.6 and 1.6 times compared to the control, respectively. Further analysis showed that riboflavin concentration in the MFC was dramatically increased from 0.1 mg/L to 1.2 mg/L by high cell density culture. Moreover, the biofilm formation on the anode surface was significantly enhanced by this new strategy. The increased accumulation of electron shuttle (riboflavin) as well as enhanced biofilm formation contributed to the improvement in anodic electrochemical activity and these factors were the underlying mechanism for MFC performance improvement by high cell density culture. This work demonstrated that high cell density culture would be a simple and practical strategy for MFC manipulation. PMID:27148719

  18. Robustness and adaptation reveal plausible cell cycle controlling subnetwork in Saccharomyces cerevisiae.

    PubMed

    Huang, Jiun-Yan; Huang, Chi-Wei; Kao, Kuo-Ching; Lai, Pik-Yin

    2013-04-10

    Biological systems are often organized spatially and temporally by multi-scale functional subsystems (modules). A specific subcellular process often corresponds to a subsystem composed of some of these interconnected modules. Accurate identification of system-level modularity organization from the large scale networks can provide valuable information on subsystem models of subcellular processes or physiological phenomena. Computational identification of functional modules from the large scale network is the key approach to solve the complexity of modularity in the past decade, but the overlapping and multi-scale nature of modules often renders unsatisfactory results in these methods. Most current methods for modularity detection are optimization-based and suffered from the drawback of size resolution limit. It is difficult to trace the origin of the unsatisfactory results, which may be due to poor data, inappropriate objective function selection or simply resulted from natural evolution, and hence no system-level accurate modular models for subcellular processes can be offered. Motivated by the idea of evolution with robustness and adaption as guiding principles, we propose a novel approach that can identify significant multi-scale overlapping modules that are sufficiently accurate at the system and subsystem levels, giving biological insights for subcellular processes. The success of our evolution strategy method is demonstrated by applying to the yeast protein-protein interaction network. Functional subsystems of important physiological phenomena can be revealed. In particular, the cell cycle controlling network is selected for detailed discussion. The cell cycle subcellular processes in yeast can be successfully dissected into functional modules of cell cycle control, cell size check point, spindle assembly checkpoint, and DNA damage check point in G2/M and S phases. The interconnections between check points and cell cycle control modules provide clues on the

  19. Accurate initiation by RNA polymerase II in a whole cell extract from Saccharomyces cerevisiae.

    PubMed

    Woontner, M; Jaehning, J A

    1990-06-01

    We have developed a simple procedure for isolating a transcriptional extract from whole yeast cells which obviates the requirement for nuclear isolation. Detection of accurate mRNA initiation by RNA polymerase II in the extract requires the use of a sensitive assay, recently described by Kornberg and co-workers (Lue, N. F., Flanagan, P. M., Sugimoto, K., and Kornberg, R. D. (1989) Science 246, 661-664) that involves activation by a GAL4-VP16 fusion protein and a template lacking guanosine residues in the coding strand. The extract is prepared from fresh or frozen yeast cells by disruption with glass beads and fractionation of proteins by ammonium sulfate precipitation. The alpha-amanitin-sensitive transcripts synthesized in the assay were identical to those produced in a parallel assay using a yeast nuclear extract. The activity of the whole cell extract is lower per mg of protein than a nuclear extract but proportional to the volume of the nucleus relative to the whole cell. The optimal ranges for several reaction components including template, mono- and divalent cations, and nucleotide substrate concentration were determined. Under optimal conditions the whole cell extract produced a maximum of approximately 1 X 10(-2) transcripts/template molecule in 30 min. PMID:2188968

  20. Re-examination of the relationship between marine virus and microbial cell abundances.

    PubMed

    Wigington, Charles H; Sonderegger, Derek; Brussaard, Corina P D; Buchan, Alison; Finke, Jan F; Fuhrman, Jed A; Lennon, Jay T; Middelboe, Mathias; Suttle, Curtis A; Stock, Charles; Wilson, William H; Wommack, K Eric; Wilhelm, Steven W; Weitz, Joshua S

    2016-01-01

    Marine viruses are critical drivers of ocean biogeochemistry, and their abundances vary spatiotemporally in the global oceans, with upper estimates exceeding 10(8) per ml. Over many years, a consensus has emerged that virus abundances are typically tenfold higher than microbial cell abundances. However, the true explanatory power of a linear relationship and its robustness across diverse ocean environments is unclear. Here, we compile 5,671 microbial cell and virus abundance estimates from 25 distinct marine surveys and find substantial variation in the virus-to-microbial cell ratio, in which a 10:1 model has either limited or no explanatory power. Instead, virus abundances are better described as nonlinear, power-law functions of microbial cell abundances. The fitted scaling exponents are typically less than 1, implying that the virus-to-microbial cell ratio decreases with microbial cell density, rather than remaining fixed. The observed scaling also implies that viral effect sizes derived from 'representative' abundances require substantial refinement to be extrapolated to regional or global scales. PMID:27572161

  1. Electricity generation in microbial fuel cells using neutral red as an electronophore

    SciTech Connect

    Park, D.H.; Zeikus, J.G.

    2000-04-01

    Neutral red (NR) was utilized as an electron mediator in microbial fuel cells consuming glucose to study both its efficiency during electricity generation and its role in altering anaerobic growth and metabolism of Escherichia coli and Actinobacillus succinogenes. A study of chemical fuel cells in which NADH, NR, and ferricyanide were the electron donor, the electronophore, and the electron acceptor, respectively, showed that electrical current produced from NADH was proportional to the concentration of NADH. Fourfold more current was produced from NADH in chemical fuel cells when NR was the electron mediator than when thionin was the electron mediator. In microbial fuel cells in which E. coli resting cells were used the amount of current produced from glucose when NR was the electron mediator was 10-fold more than the amount produced when thionin was the electron mediator. The amount of electrical energy generated and the amount of current produced from glucose in NR-mediated microbial fuel cells containing either E. coli or A. succinogenes were about 10- and 2-fold greater, respectively, when resting cells were used than when growing cells were used. Cell growth was inhibited substantially when these microbial fuel cells were making current, and more oxidized end products were formed under these conditions. When sewage sludge was used in the fuel cell, stable and equivalent levels of current were obtained with glucose, as observed in the pure-culture experiments. These results suggest that NR is better than other electron mediators used in microbial fuel cells and that sludge production can be decreased while electricity is produced in fuel cells. Their results are discussed in relation to factors that may improve the relatively low electrical efficiencies obtained with microbial fuel cells.

  2. Water structure in vitro and within Saccharomyces cerevisiae yeast cells under conditions of heat shock

    PubMed Central

    Dashnau, Jennifer L.; Conlin, Laura K.; Nelson, Hillary C. M.; Vanderkooi, Jane M.

    2008-01-01

    The OH stretch mode from water and organic hydroxyl groups have strong infrared absorption, the position of the band going to lower frequency with increased H-bonding. This band was used to study water in trehalose and glycerol solutions and in genetically modified yeast cells containing varying amounts of trehalose. Concentration-dependent changes in water structure induced by trehalose and glycerol in solution were detected, consistent with an increase of lower-energy H-bonds and interactions at the expense of higher-energy interactions. This result suggests that these molecules disrupt the water H-bond network in such a way as to strengthen molecule-water interactions while perturbing water-water interactions. The molecule-induced changes in the water H-bond network seen in solution do not translate to observable differences in yeast cells that are trehalose-deficient and trehalose-rich. Although comparison of yeast with low and high trehalose showed no observable effect on intracellular water structure, the structure of water in cells is different from that in bulk water. Cellular water exhibits a larger preference for lower-energy H-bonds or interactions over higher-energy interactions relative to that shown in bulk water. This effect is likely the result of the high concentration of biological molecules present in the cell. The ability of water to interact directly with polar groups on biological molecules may cause the preference seen for lower-energy interactions. PMID:17961925

  3. The fungicide Mancozeb induces metacaspase-dependent apoptotic cell death in Saccharomyces cerevisiae BY4741.

    PubMed

    Scariot, F J; Jahn, L M; Maianti, J P; Delamare, A P L; Echeverrigaray, S

    2016-07-01

    Mancozeb (MZ), a mixture of ethylene-bis-dithiocarbamate manganese and zinc salts, is one of the most widely used fungicides in agriculture. Toxicologic studies in mammals and mammalian cells indicate that this fungicide can cause neurological and cytological disorders, putatively associated with pro-oxidant and apoptotic effects. Yeast adaptation to sub-inhibitory concentrations of MZ has been correlated with oxidative response, proteins degradation, and energy metabolism, and its main effect on yeast has been attributed to its high reactivity with thiol groups in proteins. Herein, we show that acute MZ treatments on aerobic exponentially growing yeast of wild type (BY4741) and deletion mutant strains, coupled with multiplex flow cytometry analysis, conclusively demonstrated that MZ displays the typical features of pro-oxidant activity on Saccharomyces, elevating mitochondrial ROS, and causing hyper-polarization of mitochondrial membranes leading to apoptosis. A drastic reduction of cellular viability associated with the maintenance of cell membrane integrity, as well as phosphatidyl serine externalization on yeast cells exposed to MZ, also supports an apoptotic mode of action. Moreover, abrogation of the apoptotic response in yca1 deficient mutants indicates that metacaspase-1 is involved in the programmed cell death mechanism induced by MZ in yeast. PMID:27160815

  4. Analysis of long-term performance and microbial community structure in bio-cathode microbial desalination cells.

    PubMed

    Zhang, Huichao; Wen, Qinxue; An, Zhongyi; Chen, Zhiqiang; Nan, Jun

    2016-03-01

    A microbial desalination cell (MDC) could desalinate salt water without energy consumption and simultaneously generate bioenergy. Compared with an abiotic cathode MDC, an aerobic bio-cathode MDC is more sustainable and is less expensive to operate. In this study, the long-term operation (5500 h) performance of a bio-cathode MDC was investigated in which the power density, Coulombic efficiency, and salt removal rate were decreased by 71, 44, and 27 %, respectively. The primary reason for the system performance decrease was biofouling on the membranes, which increased internal resistance and reduced the ionic transfer and energy conversion efficiency. Changing membranes was an effective method to recover the MDC performance. The microbial community diversity in the MDC anode was low compared with that of the reported microbial fuel cell (MFC), while the abundance of Proteobacteria was 30 % higher. The content of Planctomycetes in the cathode biofilm sample was much higher than that in biofouling on the cation exchange membrane (CEM), indicating that Planctomycetes were relevant to cathode oxygen reduction. PMID:26596826

  5. Promotion of anodic electron transfer in a microbial fuel cell combined with a silicon solar cell

    NASA Astrophysics Data System (ADS)

    Ding, Hongrui; Li, Yan; Lu, Anhuai; Wang, Xin; Wang, Changqiu

    2014-05-01

    This study focuses on the promotion of electron transfer in microbial fuel cells (MFCs) by equipping a silicon solar cell (SSC) into the circuit. As compared to a sole MFC, a significant improvement of power output is observed in the MFC-SSC, that the maximum power density increases from 7.5 W m-3-19 W m-3 by 2.53 times. A linear relationship between anodic potential and current has been observed when the current is below the limiting point of SSC. We estimate the electron transfer rate can be promoted in a MFC-SSC under the condition that the anodic microbial reactions are unaffected by the incorporation of a SSC. In this way, the anodic electrons are fully pumped and enter into the external circuit. This estimation is thereby demonstrated by the 24-h test, which shows the quantity of the electrons fluent in the circuit of a MFC-SSC is doubled and the microbial oxidation efficiency is improved to 341.6% as compared with a sole MFC.

  6. Electron flux and microbial community in microbial fuel cells (open-circuit and closed-circuit modes) and fermentation.

    PubMed

    Yu, Jaecheul; Park, Youghyun; Lee, Taeho

    2015-07-01

    A closed-circuit microbial fuel cell (C-MFC) was operated to investigate the electron flux under fed-batch mode, and the results were compared to those of open-circuit MFC (O-MFC) and a fermentation reactor (F-reactor). The current was the largest electron sink (52.7% of influent SCOD) in C-MFC, whereas biomass and methane gas were the most significant electron sinks in O-MFC and F-reactor. Interestingly, some of the unknown sink may have accumulated in the electrode of O-MFC. Principal component analysis based on gradient gel electrophoresis profiles showed that the microbial communities were significantly affected by the growth conditions and the presence of electrode, regardless of the circuit connection. Therefore, the electrode and circuit mode might help to control the amount of biomass and enhance the MFC performance. PMID:25948050

  7. NADH-dependent biosensor in Saccharomyces cerevisiae: principle and validation at the single cell level

    PubMed Central

    2014-01-01

    A reporter system was constructed to measure perturbations in the NADH/NAD+ co-factor balance in yeast, by using the green fluorescent protein gene under the control of the GPD2 promoter that is induced under conditions of excess of NADH. High fluorescence levels were obtained in a glycerol 3-phosphate dehydrogenase double deletion strain (gpd1Δgpd2Δ), which is deficient in the ability to regenerate NAD+ via glycerol formation. The responsiveness of the reporter system to externally induced perturbations in NADH oxidation was also evaluated in the gpd1Δgpd2Δ strain background by addition of acetoin, as well as by introduction of a set of heterologous xylose reductases (XRs) having different selectivities for NADH. Addition of acetoin during cell proliferation under oxygen-limited conditions resulted in a more than 2-fold decrease in mean fluorescence intensity as compared to the control experiment. Strains carrying XRs with different selectivities for NADH could be distinguished at the single cell level, so that the XR with the highest selectivity for NADH displayed the lowest fluorescence. In conclusion, the designed system successfully allowed for monitoring perturbations in the cellular redox metabolism caused by environmental changes, or by heterologous gene expression. The reporter system displayed high resolution in distinguishing cytosolic NADH oxidation capacity and hence has potential to be used for high-throughput screening based on the fluorescence of single cells. PMID:25401080

  8. Stackable and submergible microbial fuel cell modules for wastewater treatment.

    PubMed

    Kim, Minsoo; Cha, Jaehwan; Yu, Jaecheul; Kim, Changwon

    2016-08-01

    The stackable and submergible microbial fuel cell (SS-MFC) system was fabricated consisting of three MFC modules (#1, #2 and #3) that were immersed in an anaerobic tank as a 30 L anode compartment. Each module consisted of the anion exchange membrane-membrane electrode assembly (A-MEA) and cation exchange membrane-MEA (C-MEA). Two MEAs shared a cathode compartment in the module and the three modules shared a anode compartment The SS-MFC system was operated with two phase. After batch feeding (phase I), the system was operated under continuous mode (phase II) with different organic concentrations (from 50 to 1000 mg/L) and different hydraulic retention times (HRT; from 3.4 to 7.2 h). The SS-MFC system successfully produced a stable voltage. A-MEA generated a lower power density than the C-MEA because of the former's high activation and resistance loss. C-MEA showed a higher average maximum power density (3.16 W/m(3)) than A-MEA (2.82 W/m(3)) at 70 mL/min (HRT of 7.2 h). The current density increased as the organic concentration was increased from 70 to 1000 mg/L in a manner consistent with Monod kinetics. When the HRT was increased from 3.4 to 7.2 h, the power densities of the C-MEAs increased from 34.3-40.9 to 40.7-45.7 mW/m(2), but those of the A-MEAs decreased from 25.3-48.0 to 27.7-40.9 mW/m(2). Although power generation was affected by HRT, organic concentrations, and separator types, the proposed SS-MFC modules can be applied to existing wastewater treatment plants. PMID:27033857

  9. Power generation from furfural using the microbial fuel cell

    NASA Astrophysics Data System (ADS)

    Luo, Yong; Liu, Guangli; Zhang, Renduo; Zhang, Cuiping

    Furfural is a typical inhibitor in the ethanol fermentation process using lignocellulosic hydrolysates as raw materials. In the literature, no report has shown that furfural can be utilized as the fuel to produce electricity in the microbial fuel cell (MFC), a device that uses microbes to convert organic compounds to generate electricity. In this study, we demonstrated that electricity was successfully generated using furfural as the sole fuel in both the ferricyanide-cathode MFC and the air-cathode MFC. In the ferricyanide-cathode MFC, the maximum power densities reached 45.4, 81.4, and 103 W m -3, respectively, when 1000 mg L -1 glucose, a mixture of 200 mg L -1 glucose and 5 mM furfural, and 6.68 mM furfural were used as the fuels in the anode solution. The corresponding Coulombic efficiencies (CE) were 4.0, 7.1, and 10.2% for the three treatments, respectively. For pure furfural as the fuel, the removal efficiency of furfural reached up to 95% within 12 h. In the air-cathode MFC using 6.68 mM furfural as the fuel, the maximum values of power density and CE were 361 mW m -2 (18 W m -3) and 30.3%, respectively, and the COD removal was about 68% at the end of the experiment (about 30 h). Increase in furfural concentrations from 6.68 to 20 mM resulted in increase in the maximum power densities from 361 to 368 mW m -2, and decrease in CEs from 30.3 to 20.6%. These results indicated that some toxic and biorefractory organics such as furfural might still be suitable resources for electricity generation using the MFC technology.

  10. Electricity production from twelve monosaccharides using microbial fuel cells

    NASA Astrophysics Data System (ADS)

    Catal, Tunc; Li, Kaichang; Bermek, Hakan; Liu, Hong

    Direct generation of electricity from monosaccharides of lignocellulosic biomass was examined using air cathode microbial fuel cells (MFCs). Electricity was generated from all carbon sources tested, including six hexoses (D-glucose, D-galactose, D(-)-levulose (fructose), L-fucose, L-rhamnose, and D-mannose), three pentoses (D-xylose, D(-)-arabinose, and D(-)-ribose), two uronic acids (D-galacturonic acid and D-glucuronic acid) and one aldonic acid (D-gluconic acid). The mixed bacterial culture, which was enriched using acetate as a carbon source, adapted well to all carbon sources tested, although the adaptation times varied from 1 to 70 h. The maximum power density obtained from these carbon sources ranged from 1240 ± 10 to 2770 ± 30 mW m -2 at current density range of 0.76-1.18 mA cm -2. D-Mannose resulted in the lowest maximum power density, whereas D-glucuronic acid generated the highest one. Coulombic efficiency ranged from 21 to 37%. For all carbon sources tested, the relationship between the maximum voltage output and the substrate concentration appeared to follow saturation kinetics at 120 Ω external resistance. The estimated maximum voltage output ranged between 0.26 and 0.44 V and half-saturation kinetic constants ranged from 111 to 725 mg L -1. Chemical oxygen demand (COD) removal was over 80% for all carbon sources tested. Results from this study indicated that lignocellulosic biomass-derived monosaccharides might be a suitable resource for electricity generation using MFC technology.

  11. Actin from Saccharomyces cerevisiae.

    PubMed Central

    Greer, C; Schekman, R

    1982-01-01

    Inhibition of DNase I activity has been used as an assay to purify actin from Saccharomyces cerevisiae (yeast actin). The final fraction, obtained after a 300-fold purification, is approximately 97% pure as judged by sodium dodecyl sulfate-gel electrophoresis. Like rabbit skeletal muscle actin, yeast actin has a molecular weight of about 43,000, forms 7-nm-diameter filaments when polymerization is induced by KCl or Mg2+, and can be decorated with a proteolytic fragment of muscle myosin (heavy meromyosin). Although heavy meromyosin ATPase activity is stimulated by rabbit muscle and yeast actins to approximately the same Vmax (2 mmol of Pi per min per mumol of heavy meromyosin), half-maximal activation (Kapp) is obtained with 14 micro M muscle actin, but requires approximately 135 micro M yeast actin. This difference suggests a low affinity of yeast actin for muscle myosin. Yeast and muscle filamentous actin respond similarly to cytochalasin and phalloidin, although the drugs have no effect on S. cerevisiae cell growth. Images PMID:6217414

  12. In vitro nonsense suppression in [psi+] and [psi-] cell-free lysates of Saccharomyces cerevisiae.

    PubMed Central

    Tuite, M F; Cox, B S; McLaughlin, C S

    1983-01-01

    An homologous in vitro assay for yeast nonsense suppressors was used to examine the effect of the cytoplasmically inherited genetic determinant [psi] on the efficiency of in vitro nonsense suppression. The efficiency of all three types of yeast tRNA-mediated nonsense suppressor (ochre, amber, and UGA) is much greater in cell-free lysates prepared from a sup+ [psi+] strain than in lysates prepared from an isogeneic sup+ [psi-] strain. Lysates prepared from a [psi-] strain, into which the [psi+] determinant was reintroduced by kar1-mediated cytoduction, support efficient suppression. Evidence is also presented that [psi-] lysates contain an inhibitor of in vitro nonsense suppression. Images PMID:6344070

  13. Trapping of DNA topoisomerase I on nick-containing DNA in cell free extracts of Saccharomyces cerevisiae.

    PubMed

    Lebedeva, Natalia; Auffret Vander Kemp, Patricia; Bjornsti, Mary-Ann; Lavrik, Olga; Boiteux, Serge

    2006-07-13

    The aim of the present study was to identify proteins that bind nicked DNA intermediates formed in the course of base excision repair (BER) in cell free extracts of Saccharomyces cerevisiae. In mammalian cells, nicks in DNA are targets of proteins such as PARP-1 or XRCC1 that have no homologues in yeast. One of the most promising methodologies to trap proteins that interact with damaged DNA lies in using a photocrosslinking technique with photoactivable dNTP analogues such as exo-N-{2-[N-(4-azido-2,5-difluoro-3-chloropyridine-6-yl)-3-aminopropionyl]-aminoethyl}-2'-deoxycytidine-5'-triphosphate (FAP-dCTP) for enzymatic synthesis of DNA probes with a photoreactive dNMP residue at the 3'-margin of a nick. Using this approach, we identified a major covalent DNA-protein adduct between a nick-containing 34-mer DNA duplex and a protein of a molecular mass of around 100-kDa. Unexpectedly, the formation of the 100-kDa adduct did not require the incorporation of the photoreactive dNMP residue at the 3'-margin of the nick nor exposure to near UV-light. However, the formation of the 100-kDa adduct strictly required a nick or a short gap in the DNA probe. Furthermore, the 100-kDa adduct was not detected in yeast extracts lacking DNA topoisomerase I (Top1). To further establish the nature of crosslinked protein, yeast Top1 was tagged with a Myc-epitope. In this case, the mobility of the Top1-DNA adduct increased by 7- kDa. Therefore, our data speak in favor of Top1 trapping by nicked DNA. In support of this hypothesis, purified yeast Top1 was also crosslinked to nicked DNA structures. Undamaged, uracil- and abasic (AP) site-containing DNAs were unable to trap Top1 under the same assay conditions. Since nicked DNA structures are frequently formed in the course of BER, their covalent linkage to Top1 has the potential to interfere with BER in vivo. PMID:16713756

  14. Saccharomyces cerevisiae Kelch Proteins and Bud14 Protein Form a Stable 520-kDa Formin Regulatory Complex That Controls Actin Cable Assembly and Cell Morphogenesis*

    PubMed Central

    Gould, Christopher J.; Chesarone-Cataldo, Melissa; Alioto, Salvatore L.; Salin, Bénédicte; Sagot, Isabelle; Goode, Bruce L.

    2014-01-01

    Formins perform essential roles in actin assembly and organization in vivo, but they also require tight regulation of their activities to produce properly functioning actin structures. Saccharomyces cerevisiae Bud14 is one member of an emerging class of formin regulators that target the FH2 domain to inhibit actin polymerization, but little is known about how these regulators are themselves controlled in vivo. Kelch proteins are critical for cell polarity and morphogenesis in a wide range of organisms, but their mechanistic roles in these processes are still largely undefined. Here, we report that S. cerevisiae Kelch proteins, Kel1 and Kel2, associate with Bud14 in cell extracts to form a stable 520-kDa complex with an apparent stoichiometry of 2:2:1 Bud14/Kel1/Kel2. Using pairwise combinations of GFP- and red fluorescent protein-tagged proteins, we show that Kel1, Kel2, and Bud14 interdependently co-localize at polarity sites. By analyzing single, double, and triple mutants, we show that Kel1 and Kel2 function in the same pathway as Bud14 in regulating Bnr1-mediated actin cable formation. Loss of any component of the complex results in long, bent, and hyper-stable actin cables, accompanied by defects in secretory vesicle traffic during polarized growth and septum formation during cytokinesis. These observations directly link S. cerevisiae Kelch proteins to the control of formin activity, and together with previous observations made for S. pombe homologues tea1p and tea3p, they have broad implications for understanding Kelch function in other systems. PMID:24828508

  15. Defect in cell wall integrity of the yeast saccharomyces cerevisiae caused by a mutation of the GDP-mannose pyrophosphorylase gene VIG9.

    PubMed

    Yoda, K; Kawada, T; Kaibara, C; Fujie, A; Abe, M; Hitoshi; Hashimoto; Shimizu, J; Tomishige, N; Noda, Y; Yamasaki, M

    2000-09-01

    The Saccharomyces cerevisiae VIG9 gene encodes GDP-mannose pyrophosphorylase, which synthesizes GDP-mannose from GTP and mannose-1-phosphate. Although the null mutant was lethal, the vig9 mutants so far obtained showed no growth defect but immature protein glycosylation and drug hypersensitivity. During our search for cell-wall mutants, we found a novel temperature-sensitive mutant, JS30, which required an osmotic stabilizer for viability. JS30 excreted cell surface proteins in the medium without any indication of cell lysis. Although conventional genetic analysis using mating was impossible, by detailed characterization of JS30 including an in vitro enzyme assay and nucleotide sequencing, we found the defect of JS30 was due to a mutation in the VIG9 gene. These results indicated a critical role of GDP-mannose in maintenance of cell-wall integrity. PMID:11055399

  16. High-cell-density fermentation for ergosterol production by Saccharomyces cerevisiae.

    PubMed

    Shang, Fei; Wen, Shaohong; Wang, Xi; Tan, Tianwei

    2006-01-01

    The direct feedback control of glucose using an on-line ethanol concentration monitor for ergosterol production by high-cell-density fermentation was investigated and the fermentation parameters (e.g., pH, dissolved oxygen, ethanol concentration, oxygen uptake rate, carbon dioxide evolution rate and respiratory quotient) were analyzed. Controlling glucose feeding rate in accordance with ethanol concentration and adjusting pH with ammonia during the fermentation process were effective fed-batch methods for ergosterol production. The fermentation parameters well described the variation of the whole fermentation process. Cultivation in a 5 l fermentor was carried out under the following conditions: culture temperature, 30 degrees C; pH, 5.5; agitation speed, 600 rpm; fermentation time, 60 h; controlling ethanol concentration below 1% and keeping respiratory quotient (RQ) at approximately 1.0. Under these conditions, the yeast dry weight reached 120 g/l and the ergosterol yield reached 1500 mg/l. PMID:16503289

  17. Searches for Microbial Cells with Fluorescence Loggers with Single-cell Sensitivity

    NASA Astrophysics Data System (ADS)

    Price, P. B.; Rohde, R. A.; Bay, R. C.

    2007-12-01

    Two known habitats for microbial metabolism in ice are surfaces of mineral grains and liquid veins along three- grain boundaries. Several problems suggest the need for a third habitat: veins usually contain toxic liquid; some microorganisms are too large to fit into a vein; veins may not be present at all depths; and the oxygen concentration in veins does not permit the coexistence of both strict anaerobes and aerobes in the same region. We show that a more general habitat avoids these problems. Isolated microbes frozen in ice and not in contact with a vein or grain can metabolize by redox reactions with dissolved small molecules diffusing through the ice lattice. The two requirements are that the gaseous reactants have sufficiently high equilibrium concentrations and diffusion coefficients to provide enough metabolic energy to repair macromolecular damage as it occurs. Molecules with less than ~6 atoms (e.g., H2, O2, N2¬, CO, CO2, CH4, H2S, NH3, HNO3, HCHO, and HCOOH) have values of diffusion coefficient D(T) that exceed ~10- 15 m2 s-1, which is sufficient to sustain microbial life in ice. For terrestrial environments, we show that there is an adequate supply of such molecules diffusing throughout deep glacial ice to sustain metabolism for millions of years. Our recent noninvasive observations of ice cores from GISP2 and WAIS Divide provide evidence for this habitat. Using scanning fluorimetry to map proteins (a proxy for cells) and F420 (a proxy for methanogens) in ice cores, we find isolated spikes of fluorescence consistent with as few as one microbial cell in a volume 0.16 microliter with the protein mapper and in 1.9 microliter with the methanogen mapper. With such precise localization one could use a nanomanipulator to extract single cells for molecular identification. Low- power, miniaturized versions of these instruments could search for single cells in subglacial lakes, Martian ice- rich permafrost, and Europan ice.

  18. Metabolic pathway engineering for fatty acid ethyl ester production in Saccharomyces cerevisiae using stable chromosomal integration.

    PubMed

    de Jong, Bouke Wim; Shi, Shuobo; Valle-Rodríguez, Juan Octavio; Siewers, Verena; Nielsen, Jens

    2015-03-01

    Fatty acid ethyl esters are fatty acid derived molecules similar to first generation biodiesel (fatty acid methyl esters; FAMEs) which can be produced in a microbial cell factory. Saccharomyces cerevisiae is a suitable candidate for microbial large scale and long term cultivations, which is the typical industrial production setting for biofuels. It is crucial to conserve the metabolic design of the cell factory during industrial cultivation conditions that require extensive propagation. Genetic modifications therefore have to be introduced in a stable manner. Here, several metabolic engineering strategies for improved production of fatty acid ethyl esters in S. cerevisiae were combined and the genes were stably expressed from the organisms' chromosomes. A wax ester synthase (ws2) was expressed in different yeast strains with an engineered acetyl-CoA and fatty acid metabolism. Thus, we compared expression of ws2 with and without overexpression of alcohol dehydrogenase (ADH2), acetaldehyde dehydrogenase (ALD6) and acetyl-CoA synthetase (acs SE (L641P) ) and further evaluated additional overexpression of a mutant version of acetyl-CoA decarboxylase (ACC1 (S1157A,S659A) ) and the acyl-CoA binding protein (ACB1). The combined engineering efforts of the implementation of ws2, ADH2, ALD6 and acs SE (L641P) , ACC1 (S1157A,S659A) and ACB1 in a S. cerevisiae strain lacking storage lipid formation (are1Δ, are2Δ, dga1Δ and lro1Δ) and β-oxidation (pox1Δ) resulted in a 4.1-fold improvement compared with sole expression of ws2 in S. cerevisiae. PMID:25422103

  19. Transcriptional profiling of Saccharomyces cerevisiae T2 cells upon exposure to hardwood spent sulphite liquor: comparison to acetic acid, furfural and hydroxymethylfurfural.

    PubMed

    Bajwa, Paramjit K; Ho, Chi-Yip; Chan, Chi-Kin; Martin, Vincent J J; Trevors, Jack T; Lee, Hung

    2013-06-01

    Global gene expression was analyzed in Saccharomyces cerevisiae T2 cells grown in the presence of hardwood spent sulphite liquor (HW SSL) and each of the three main inhibitors in HW SSL, acetic acid, hydroxymethyfurfural (HMF) and furfural, using a S. cerevisiae DNA oligonucleotide microarray. The objective was to compare the gene expression profiles of T2 cells in response to the individual inhibitors against that elicited in response to HW SSL. Acetic acid mainly affected the expression of genes related to the uptake systems of the yeast as well as energy generation and metabolism. Furfural and HMF mainly affected the transcription of genes involved in the redox balance of the cell. On the other hand, the effect of HW SSL on S. cerevisiae T2 cells was distinct and considerably more diverse as compared to the effect of individual inhibitors found in lignocellulosic hydrolysates. This is not surprising as HW SSL contains a complex mixture of inhibitors which may act synergistically. HW SSL elicited significant changes in expression of genes involved in diverse and multiple effects on several aspects of the cellular structure and function. A notable response to HW SSL was decreased expression of the ribosomal protein genes in T2 cells. In addition, HW SSL decreased the expression of genes functioning in the synthesis and transport of proteins as well as metabolism of carbohydrates, lipids, vitamins and vacuolar proteins. Furthermore, the expression of genes involved in multidrug resistance, iron transport and pheromone response was increased, suggesting that T2 cells grown in the presence of HW SSL may have activated pheromone response and/or activated pleiotropic drug response. Some of the largest changes in gene expression were observed in the presence of HW SSL and the affected genes are involved in mating, iron transport, stress response and phospholipid metabolism. A total of 59 out of the 400 genes differentially expressed in the presence of HW SSL, acetic

  20. Modular Spectral Imaging System for Discrimination of Pigments in Cells and Microbial Communities▿ †

    PubMed Central

    Polerecky, Lubos; Bissett, Andrew; Al-Najjar, Mohammad; Faerber, Paul; Osmers, Harald; Suci, Peter A.; Stoodley, Paul; de Beer, Dirk

    2009-01-01

    Here we describe a spectral imaging system for minimally invasive identification, localization, and relative quantification of pigments in cells and microbial communities. The modularity of the system allows pigment detection on spatial scales ranging from the single-cell level to regions whose areas are several tens of square centimeters. For pigment identification in vivo absorption and/or autofluorescence spectra are used as the analytical signals. Along with the hardware, which is easy to transport and simple to assemble and allows rapid measurement, we describe newly developed software that allows highly sensitive and pigment-specific analyses of the hyperspectral data. We also propose and describe a number of applications of the system for microbial ecology, including identification of pigments in living cells and high-spatial-resolution imaging of pigments and the associated phototrophic groups in complex microbial communities, such as photosynthetic endolithic biofilms, microbial mats, and intertidal sediments. This system provides new possibilities for studying the role of spatial organization of microorganisms in the ecological functioning of complex benthic microbial communities or for noninvasively monitoring changes in the spatial organization and/or composition of a microbial community in response to changing environmental factors. PMID:19074609

  1. Modular spectral imaging system for discrimination of pigments in cells and microbial communities.

    PubMed

    Polerecky, Lubos; Bissett, Andrew; Al-Najjar, Mohammad; Faerber, Paul; Osmers, Harald; Suci, Peter A; Stoodley, Paul; de Beer, Dirk

    2009-02-01

    Here we describe a spectral imaging system for minimally invasive identification, localization, and relative quantification of pigments in cells and microbial communities. The modularity of the system allows pigment detection on spatial scales ranging from the single-cell level to regions whose areas are several tens of square centimeters. For pigment identification in vivo absorption and/or autofluorescence spectra are used as the analytical signals. Along with the hardware, which is easy to transport and simple to assemble and allows rapid measurement, we describe newly developed software that allows highly sensitive and pigment-specific analyses of the hyperspectral data. We also propose and describe a number of applications of the system for microbial ecology, including identification of pigments in living cells and high-spatial-resolution imaging of pigments and the associated phototrophic groups in complex microbial communities, such as photosynthetic endolithic biofilms, microbial mats, and intertidal sediments. This system provides new possibilities for studying the role of spatial organization of microorganisms in the ecological functioning of complex benthic microbial communities or for noninvasively monitoring changes in the spatial organization and/or composition of a microbial community in response to changing environmental factors. PMID:19074609

  2. Series assembly of microbial desalination cells containing stacked electrodialysis cells for partial or complete seawater desalination.

    PubMed

    Kim, Younggy; Logan, Bruce E

    2011-07-01

    A microbial desalination cell (MDC) is a new approach for desalinating water based on using the electrical current generated by exoelectrogenic bacteria. Previously developed MDCs have used only one or two desalination chambers with substantial internal resistance, and used low salinity catholytes containing a buffered or acid solution. Here we show that substantially improved MDC performance can be obtained even with a nonbuffered, saline catholyte, by using an electrodialysis stack consisting of 5 pairs of desalting and concentrating cells. When 4 stacked MDCs were used in series (20 total pairs of desalination chambers), the salinity of 0.06 L of synthetic seawater (35 g/L NaCl) was reduced by 44% using 0.12 L of anode solution (2:1). The resistive loss in the electrodialysis stack was negligible due to minimization of the intermembrane distances, and therefore the power densities produced by the MDC were similar to those produced by single chamber microbial fuel cells (MFCs) lacking desalination chambers. The observed current efficiency was 86%, indicating separation of 4.3 pairs of sodium and chloride ions for every electron transferred through the circuit. With two additional stages (total of 3.8 L of anolyte), desalination was increased to 98% salt removal, producing 0.3 L of fresh water (12.6:1). These results demonstrate that stacked MDCs can be used for efficient desalination of seawater while at the same time achieving power densities comparable to those obtained in MFCs. PMID:21671676

  3. Riboflavin-shuttled extracellular electron transfer from Enterococcus faecalis to electrodes in microbial fuel cells.

    PubMed

    Zhang, Enren; Cai, Yamin; Luo, Yue; Piao, Zhe

    2014-11-01

    Great attention has been focused on Gram-negative bacteria in the application of microbial fuel cells. In this study, the Gram-positive bacterium Enterococcus faecalis was employed in microbial fuel cells. Bacterial biofilms formed by E. faecalis ZER6 were investigated with respect to electricity production through the riboflavin-shuttled extracellular electron transfer. Trace riboflavin was shown to be essential for transferring electrons derived from the oxidation of glucose outside the peptidoglycan layer in the cell wall of E. faecalis biofilms formed on the surface of electrodes, in the absence of other potential electron mediators (e.g., yeast extract). PMID:25345758

  4. Role of type I interferons in inflammasome activation, cell death, and disease during microbial infection

    PubMed Central

    Malireddi, R. K. Subbarao; Kanneganti, Thirumala-Devi

    2013-01-01

    Interferons (IFNs) were discovered over a half-century ago as antiviral factors. The role of type I IFNs has been studied in the pathogenesis of both acute and chronic microbial infections. Deregulated type I IFN production results in a damaging cascade of cell death, inflammation, and immunological host responses that can lead to tissue injury and disease progression. Here, we summarize the role of type I IFNs in the regulation of cell death and disease during different microbial infections, ranging from viruses and bacteria to fungal pathogens. Understanding the specific mechanisms driving type I IFN-mediated cell death and disease could aid in the development of targeted therapies. PMID:24273750

  5. [Advance in the bioavailability monitoring of heavy metal based on microbial whole-cell sensor].

    PubMed

    Hou, Qi-Hui; Ma, An-Shou; Zhuang, Xiu-Liang; Zhuang, Guo-Qiang

    2013-01-01

    Microbial whole-cell biosensor is an excellent tool to assess the bioavailability of heavy metal in soil and water. However, the traditional physicochemical instruments are applied to detect the total metal. Furthermore, microbial whole-cell biosensor is simple, rapid and economical in manipulating, and is thus a highly qualified candidate for emergency detection of pollution incidents. The biological component of microbial whole-cell biosensor mostly consists of metalloregulatory proteins and reporter genes. In detail, metalloregulatory proteins mainly include the MerR family, ArsR family and RS family, and reporter genes mainly include gfp, lux and luc. Metalloregulatory protein and reporter gene are related to the sensitivity, specificity and properties in monitoring. The bioavailability of heavy metals is alterable under different conditions, influenced by pH, chelate and detection methods and so on. Increasing the accumulation of intracellular heavy metal, modifying the metalloregulatory proteins and optimizing the detecting conditions are important for improving the sensitivity, specificity and accuracy of the microbial whole-cell biosensor. The future direction of microbial whole-cell biosensor is to realize the monitoring of pollutions in situ and on line. PMID:23487961

  6. Minimal RED cell pairs markedly improve electrode kinetics and power production in microbial reverse electrodialysis cells.

    PubMed

    Cusick, Roland D; Hatzell, Marta; Zhang, Fang; Logan, Bruce E

    2013-12-17

    Power production from microbial reverse electrodialysis cell (MRC) electrodes is substantially improved compared to microbial fuel cells (MFCs) by using ammonium bicarbonate (AmB) solutions in multiple RED cell pair stacks and the cathode chamber. Reducing the number of RED membranes pairs while maintaining enhanced electrode performance could help to reduce capital costs. We show here that using only a single RED cell pair (CP), created by operating the cathode in concentrated AmB, dramatically increased power production normalized to cathode area from both acetate (Acetate: from 0.9 to 3.1 W/m(2)-cat) and wastewater (WW: 0.3 to 1.7 W/m(2)), by reducing solution and charge transfer resistances at the cathode. A second RED cell pair increased RED stack potential and reduced anode charge transfer resistance, further increasing power production (Acetate: 4.2 W/m(2); WW: 1.9 W/m(2)). By maintaining near optimal electrode power production with fewer membranes, power densities normalized to total membrane area for the 1-CP (Acetate: 3.1 W/m(2)-mem; WW: 1.7 W/m(2)) and 2-CP (Acetate: 1.3 W/m(2)-mem; WW: 0.6 W/m(2)) reactors were much higher than previous MRCs (0.3-0.5 W/m(2)-mem with acetate). While operating at peak power, the rate of wastewater COD removal, normalized to reactor volume, was 30-50 times higher in 1-CP and 2-CP MRCs than that in a single chamber MFC. These findings show that even a single cell pair AmB RED stack can significantly enhance electrical power production and wastewater treatment. PMID:24224718

  7. Size and Carbon Content of Sub-seafloor Microbial Cells at Landsort Deep, Baltic Sea

    PubMed Central

    Braun, Stefan; Morono, Yuki; Littmann, Sten; Kuypers, Marcel; Aslan, Hüsnü; Dong, Mingdong; Jørgensen, Bo B.; Lomstein, Bente Aa.

    2016-01-01

    The discovery of a microbial ecosystem in ocean sediments has evoked interest in life under extreme energy limitation and its role in global element cycling. However, fundamental parameters such as the size and the amount of biomass of sub-seafloor microbial cells are poorly constrained. Here we determined the volume and the carbon content of microbial cells from a marine sediment drill core retrieved by the Integrated Ocean Drilling Program (IODP), Expedition 347, at Landsort Deep, Baltic Sea. To determine their shape and volume, cells were separated from the sediment matrix by multi-layer density centrifugation and visualized via epifluorescence microscopy (FM) and scanning electron microscopy (SEM). Total cell-carbon was calculated from amino acid-carbon, which was analyzed by high-performance liquid chromatography (HPLC) after cells had been purified by fluorescence-activated cell sorting (FACS). The majority of microbial cells in the sediment have coccoid or slightly elongated morphology. From the sediment surface to the deepest investigated sample (~60 m below the seafloor), the cell volume of both coccoid and elongated cells decreased by an order of magnitude from ~0.05 to 0.005 μm3. The cell-specific carbon content was 19–31 fg C cell−1, which is at the lower end of previous estimates that were used for global estimates of microbial biomass. The cell-specific carbon density increased with sediment depth from about 200 to 1000 fg C μm−3, suggesting that cells decrease their water content and grow small cell sizes as adaptation to the long-term subsistence at very low energy availability in the deep biosphere. We present for the first time depth-related data on the cell volume and carbon content of sedimentary microbial cells buried down to 60 m below the seafloor. Our data enable estimates of volume- and biomass-specific cellular rates of energy metabolism in the deep biosphere and will improve global estimates of microbial biomass.

  8. Characterization and optimization of cathodic conditions for H2O2 synthesis in microbial electrochemical cells

    EPA Science Inventory

    Cathode potential and O2 supply methods were investigated to improve H2O2 synthesis in an electrochemical cell, and optimal cathode conditions were applied for microbial electrochemical cells (MECs). Using aqueous O2 for the cathode significantly improved current density, but H2...

  9. Peptidase activities in Saccharomyces cerevisiae.

    PubMed Central

    Rose, B; Becker, J M; Naider, F

    1979-01-01

    At least four distinct aminopeptidase activities and a single dipeptidase activity were found in cell extracts of a leucine-lysine auxotroph of Saccharomyces cerevisiae. The assay for peptidase activity involved polyacrylamide gel electrophoresis followed by an enzyme-coupled activity staining procedure. The aminopeptidases had largely overlapping specificities but could be distinguished from one another by their electrophoretic mobilities and activities toward different peptide substrates. Substrates tested included both free and blocked di- and tripeptides and amino acid derivatives. Images PMID:378955

  10. Wiring microbial biofilms to the electrode by osmium redox polymer for the performance enhancement of microbial fuel cells.

    PubMed

    Yuan, Yong; Shin, Hyosul; Kang, Chan; Kim, Sunghyun

    2016-04-01

    An osmium redox polymer, PAA-PVI-[Os(4,4'-dimethyl-2,2'-bipyridine)2Cl]+/2+ that has been used in enzymatic fuel cells and microbial sensors, was applied for the first time to the anode of single-chamber microbial fuel cells with the mixed culture inoculum aiming at enhancing performance. Functioning as a molecular wire connecting the biofilm to the anode, power density increased from 1479 mW m(-2) without modification to 2355 mW m(-2) after modification of the anode. Evidence from cyclic voltammetry showed that the catalytic activity of an anodic biofilm was greatly enhanced in the presence of an osmium redox polymer, indicating that electrons were more efficiently transferred to the anode via co-immobilized osmium complex tethered to wiring polymer chains at the potential range of -0.3 V-+0.1 V (vs. SCE). The optimum amount of the redox polymer was determined to be 0.163 mg cm(-2). PMID:26599210

  11. Immobilization of microbial cell and yeast cell and its application to biomass conversion using radiation techniques

    NASA Astrophysics Data System (ADS)

    Kaetsu, Isao; Kumakura, Minoru; Fujimura, Takashi; Kasai, Noboru; Tamada, Masao

    The recent results of immobilization of cellulase-producing cells and ethanol-fermentation yeast by radiation were reported. The enzyme of cellulase produced by immobilized cells was used for saccharification of lignocellulosic wastes and immobilized yeast cells were used for fermentation reaction from glucose to ethanol. The wastes such as chaff and bagasse were treated by γ-ray or electron-beam irradiation in the presence of alkali and subsequent mechanical crushing, to form a fine powder less than 50 μm in diameter. On the other hand, Trichoderma reesei as a cellulase-producing microbial cell was immobilized on a fibrous carrier having a specific porous structure and cultured to produce cellulase. The enzymatic saccharification of the pretreated waste was carried out using the produced cellulase. The enhanced fermentation process to produce ethanol from glucose with the immobilized yeast by radiation was also studied. The ethanol productivity of immobilized growing yeast cells thus obtained was thirteen times that of free yeast cells in a 1:1 volume of liquid medium to immobilized yeast cells.

  12. Live Cell Discovery of Microbial Vitamin Transport and Enzyme-Cofactor Interactions.

    PubMed

    Anderson, Lindsey N; Koech, Phillip K; Plymale, Andrew E; Landorf, Elizabeth V; Konopka, Allan; Collart, Frank R; Lipton, Mary S; Romine, Margaret F; Wright, Aaron T

    2016-02-19

    The rapid completion of microbial genomes is inducing a conundrum in functional gene discovery. Novel methods are needed to shorten the gap between characterizing a microbial genome and experimentally validating bioinformatically predicted functions. Of particular importance are transport mechanisms, which shuttle nutrients such as B vitamins and metabolites across cell membranes and are required for the survival of microbes ranging from members of environmental microbial communities to pathogens. Methods to accurately assign function and specificity for a wide range of experimentally unidentified and/or predicted membrane-embedded transport proteins, along with characterization of intracellular enzyme-cofactor associations, are needed to enable a significantly improved understanding of microbial biochemistry and physiology, microbial interactions, and microbial responses to perturbations. Chemical probes derived from B vitamins B1, B2, and B7 have allowed us to experimentally address the aforementioned needs by identifying B vitamin transporters and intracellular enzyme-cofactor associations through live cell labeling of the filamentous anoxygenic photoheterotroph, Chloroflexus aurantiacus J-10-fl, known to employ mechanisms for both B vitamin biosynthesis and environmental salvage. Our probes provide a unique opportunity to directly link cellular activity and protein function back to ecosystem and/or host dynamics by identifying B vitamin transport and cofactor-dependent interactions required for survival. PMID:26669591

  13. Application of a weak magnetic field to improve microbial fuel cell performance.

    PubMed

    Tong, Zhong-Hua; Yu, Han-Qing; Li, Wen-Wei; Wang, Yun-Kun; Sun, Min; Liu, Xian-Wei; Sheng, Guo-Ping

    2015-12-01

    Microbial fuel cells (MFCs) have emerged as a promising technology for wastewater treatment with concomitant energy production but the performance is usually limited by low microbial activities. This has spurred intensive research interest for microbial enhancement. This study demonstrated an interesting stimulation effect of a static magnetic field (MF) on sludge-inoculated MFCs and explored into the mechanisms. The implementation of a 100-mT MF accelerated the reactor startup and led to increased electricity generation. Under the MF exposure, the activation loss of the MFC was decreased, but there was no increased secretion of redox mediators. Thus, the MF effect was mainly due to enhanced bioelectrochemical activities of anodic microorganisms, which are likely attributed to the oxidative stress and magnetohydrodynamic effects under an MF exposure. This work implies that weak MF may be applied as a simple and effective approach to stimulate microbial activities for various bioelectrochemical energy production and decontamination applications. PMID:26410373

  14. Enhanced biofilm distribution and cell performance of microfluidic microbial fuel cells with multiple anolyte inlets.

    PubMed

    Yang, Yang; Ye, Dingding; Liao, Qiang; Zhang, Pengqing; Zhu, Xun; Li, Jun; Fu, Qian

    2016-05-15

    A laminar-flow controlled microfluidic microbial fuel cell (MMFC) is considered as a promising approach to be a bio-electrochemical system (BES). But poor bacterial colonization and low power generation are two severe bottlenecks to restrict its development. In this study, we reported a MMFC with multiple anolyte inlets (MMFC-MI) to enhance the biofilm formation and promote the power density of MMFCs. Voltage profiles during the inoculation process demonstrated MMFC-MI had a faster start-up process than the conventional microfluidic microbial fuel cell with one inlet (MMFC-OI). Meanwhile, benefited from the periodical replenishment of boundary layer near the electrode, a more densely-packed bacterial aggregation was observed along the flow direction and also the substantially low internal resistance for MMFC-MI. Most importantly, the output power density of MMFC-MI was the highest value among the reported µl-scale MFCs to our best knowledge. The presented MMFC-MI appears promising for bio-chip technology and extends the scope of microfluidic energy. PMID:26735875

  15. A new cultivation method for microbial oil production: cell pelletization and lipid accumulation by Mucor circinelloides

    PubMed Central

    2011-01-01

    The recent energy crisis has triggered significant attention on the microbial synthesis of lipids, which comprise the raw material for biodiesel production. Microbial oil accumulation with filamentous fungi has great potential because filamentous fungi can form pellets during cell growth, and these pellets are much easier to harvest from cell broth. This paper focuses on the cell pelletization process of the oleaginous Mucor circinelloides. We have studied the effect of various cultural conditions on pelletized cell growth and lipid accumulation. This study is the first to report that pH adjustment during cell growth plays a key role in pellet formation of M. circinelloides and describes a handy method by which to induce cell pelletization in submerged fungal cultivation. Our study reveals that cell growth and lipid production are not significantly affected by pelletization and that lipid accumulation is triggered at stressed conditions, such as a high carbon-to-nitrogen ratio and high temperature. PMID:21635739

  16. Variable Cell Morphology Approach for Individual-Based Modeling of Microbial Communities

    PubMed Central

    Storck, Tomas; Picioreanu, Cristian; Virdis, Bernardino; Batstone, Damien J.

    2014-01-01

    An individual-based, mass-spring modeling framework has been developed to investigate the effect of cell properties on the structure of biofilms and microbial aggregates through Lagrangian modeling. Key features that distinguish this model are variable cell morphology described by a collection of particles connected by springs and a mechanical representation of deformable intracellular, intercellular, and cell-substratum links. A first case study describes the colony formation of a rod-shaped species on a planar substratum. This case shows the importance of mechanical interactions in a community of growing and dividing rod-shaped cells (i.e., bacilli). Cell-substratum links promote formation of mounds as opposed to single-layer biofilms, whereas filial links affect the roundness of the biofilm. A second case study describes the formation of flocs and development of external filaments in a mixed-culture activated sludge community. It is shown by modeling that distinct cell-cell links, microbial morphology, and growth kinetics can lead to excessive filamentous proliferation and interfloc bridging, possible causes for detrimental sludge bulking. This methodology has been extended to more advanced microbial morphologies such as filament branching and proves to be a very powerful tool in determining how fundamental controlling mechanisms determine diverse microbial colony architectures. PMID:24806936

  17. Substrates and pathway of electricity generation in a nitrification-based microbial fuel cell.

    PubMed

    Chen, Hui; Zheng, Ping; Zhang, Jiqiang; Xie, Zuofu; Ji, Junyuan; Ghulam, Abbas

    2014-06-01

    Nitrification-based microbial fuel cell (N-MFC) is a novel inorganic microbial fuel cell based on nitrification in the anode compartment. So far, little information is available on the substrates and pathway of N-MFC. The results of this study indicated that apart from the primary nitrification substrate (ammonium), the intermediates (hydroxylamine and nitrite) could also serve as anodic fuel to generate current, and the end product nitrate showed an inhibitory effect on electricity generation. Based on the research, a pathway of electricity generation was proposed for N-MFC: ammonium was oxidized first to nitrite by ammonia-oxidizing bacteria (AOB), then the nitrite in anolyte and the potassium permanganate in catholyte constituted a chemical cell to generate current. In other words, the electricity generation in N-MFC was not only supported by microbial reaction as we expected, but both biological and electrochemical reactions contributed. PMID:24704886

  18. Bioconversion of cellulose into electrical energy in microbial fuel cells

    NASA Astrophysics Data System (ADS)

    Rismani-Yazdi, Hamid

    In microbial fuel cells (MFCs), bacteria generate electricity by mediating the oxidation of organic compounds and transferring the resulting electrons to an anode electrode. The first objective of this study was to test the possibility of generating electricity with rumen microorganisms as biocatalysts and cellulose as the electron donor in two-compartment MFCs. Maximum power density reached 55 mW/m2 (1.5 mA, 313 mV) with cellulose as the electron donor. Cellulose hydrolysis and electrode reduction were shown to support the production of current. The electrical current was sustained for over two months with periodic cellulose addition. Clarified rumen fluid and a soluble carbohydrate mixture, serving as the electron donors, could also sustain power output. The second objective was to analyze the composition of the bacterial communities enriched in the cellulose-fed MFCs. Denaturing gradient gel electrophoresis of PCR amplified 16S rRNA genes revealed that the microbial communities differed when different substrates were used in the MFCs. The anode-attached and the suspended consortia were shown to be different within the same MFC. Cloning and analysis of 16S rRNA gene sequences indicated that the most predominant bacteria in the anode-attached consortia were related to Clostridium spp., while Comamonas spp. was abundant in the suspended consortia. The external resistance affects the characteristic outputs of MFCs by controlling the flow of electrons from the anode to the cathode. The third objective of this study was to determine the effect of various external resistances on power output and coulombic efficiency of cellulose-fed MFCs. Four external resistances (20, 249, 480, and 1000 ohms) were tested with a systematic approach of operating parallel MFCs independently at constant circuit loads for three months. A maximum power density of 66 mWm-2 was achieved by MFCs with 20 ohms circuit load, while MFCs with 249, 480 and1000 ohms external resistances produced 57

  19. Mutations in the Saccharomyces Cerevisiae Type 2a Protein Phosphatase Catalytic Subunit Reveal Roles in Cell Wall Integrity, Actin Cytoskeleton Organization and Mitosis

    PubMed Central

    Evans, DRH.; Stark, MJR.

    1997-01-01

    Temperature-sensitive mutations were generated in the Saccharomyces cerevisiae PPH22 gene that, together with its homologue PPH21, encode the catalytic subunit of type 2A protein phosphatase (PP2A). At the restrictive temperature (37°), cells dependent solely on pph22(ts) alleles for PP2A function displayed a rapid arrest of proliferation. Ts(-) pph22 mutant cells underwent lysis at 37°, showing an accompanying viability loss that was suppressed by inclusion of 1 M sorbitol in the growth medium. Ts(-) pph22 mutant cells also displayed defects in bud morphogenesis and polarization of the cortical actin cytoskeleton at 37°. PP2A is therefore required for maintenance of cell integrity and polarized growth. On transfer from 24° to 37°, Ts(-) pph22 mutant cells accumulated a 2N DNA content indicating a cell cycle block before completion of mitosis. However, during prolonged incubation at 37°, many Ts(-) pph22 mutant cells progressed through an aberrant nuclear division and accumulated multiple nuclei. Ts(-) pph22 mutant cells also accumulated aberrant microtubule structures at 37°, while under semi-permissive conditions they were sensitive to the microtubule-destabilizing agent benomyl, suggesting that PP2A is required for normal microtubule function. Remarkably, the multiple defects of Ts(-) pph22 mutant cells were suppressed by a viable allele (SSD1-v1) of the polymorphic SSD1 gene. PMID:9071579

  20. Saccharomyces cerevisiae SHSY detoxifies petroleum n-alkanes by an induced CYP52A58 and an enhanced order in cell surface hydrophobicity.

    PubMed

    Hanano, Abdulsamie; Shaban, Mouhnad; Almousally, Ibrahem; Al-Ktaifani, Mahmoud

    2015-09-01

    Environmental hydrocarbon contamination has a serious hazard to human health. Alkanes, the major component of hydrocarbons, can be consumed by various species of yeast. We previously identified a new strain SHSY of Saccharomyces cerevisiae with a remarkable ability to utilize the petroleum crude-oil (PCO) in aqueous solution. The current study demonstrated that the n-alkanes-assimilation activity of S. cerevisiae SHSY was related to an induced microsomal protein of 59 kDa approximately. The identified ORF encoded a protein of 517 amino acids and shared 93% sequence identity with an alkane-inducible hydroxylase CYP52A53 isolated from Scheffersomyces stipitis CBS. It was therefore referred as CYP52A58. The catalytic activity of the recombinant CYP52A58 was confirmed by the hydroxylation of n-alkanes, it showed an optimal mono-terminal hydroxylation activity toward n-hexadecane. Moreover, the ability of the yeast to use n-alkanes was accompanied with an increasing level in cell wall mannoproteins. Two differential protein bands were detected in the mannoproteins extracted from PCO-grown yeast. In parallel, a significant increase in the fatty acids content with a high degree of unsaturation was subsequently detected in the PCO-grown yeast. This study characterizes a safe and potential microorganism to remove n-alkanes from the aquatic environment. PMID:25434275

  1. Effect of bacterial cell size on electricity generation in a single-compartmented microbial fuel cell.

    PubMed

    Lee, Seung Won; Jeon, Bo Young; Park, Doo Hyun

    2010-04-01

    A single-compartmented microbial fuel cell composed of a graphite felt anode modified with Neutral Red (NR-anode) and a porous Fe(II)-carbon cathode (FeC-cathode) were compared for electricity generation from Microbacterium sp. and Pseudomonas sp. under identical conditions. Pseudomonas sp. was more than four times the size of Microbacterium sp. based on SEM images. In cyclic voltammetry, the redox reaction between Microbacterium sp and electrode was three times the rate observed between Pseudomonas sp. and the electrode based on the Y-axis (current) variation of cyclic voltammogram. The electric power generated by Microbacterium sp. was approx 3-4 times higher than that with Pseudomonas sp. during incubation for more than 150 days in the fuel cell. PMID:20013300

  2. Plasma membrane/cell wall perturbation activates a novel cell cycle checkpoint during G1 in Saccharomyces cerevisiae.

    PubMed

    Kono, Keiko; Al-Zain, Amr; Schroeder, Lea; Nakanishi, Makoto; Ikui, Amy E

    2016-06-21

    Cellular wound healing or the repair of plasma membrane/cell wall damage (plasma membrane damage) occurs frequently in nature. Although various cellular perturbations, such as DNA damage, spindle misalignment, and impaired daughter cell formation, are monitored by cell cycle checkpoint mechanisms in budding yeast, whether plasma membrane damage is monitored by any of these checkpoints remains to be addressed. Here, we define the mechanism by which cells sense membrane damage and inhibit DNA replication. We found that the inhibition of DNA replication upon plasma membrane damage requires GSK3/Mck1-dependent degradation of Cdc6, a component of the prereplicative complex. Furthermore, the CDK inhibitor Sic1 is stabilized in response to plasma membrane damage, leading to cell integrity maintenance in parallel with the Mck1-Cdc6 pathway. Cells defective in both Cdc6 degradation and Sic1 stabilization failed to grow in the presence of plasma membrane damage. Taking these data together, we propose that plasma membrane damage triggers G1 arrest via Cdc6 degradation and Sic1 stabilization to promote the cellular wound healing process. PMID:27274080

  3. Tracking heavy water (D2O) incorporation for identifying and sorting active microbial cells

    PubMed Central

    Berry, David; Mader, Esther; Lee, Tae Kwon; Woebken, Dagmar; Wang, Yun; Zhu, Di; Palatinszky, Marton; Schintlmeister, Arno; Schmid, Markus C.; Hanson, Buck T.; Shterzer, Naama; Mizrahi, Itzhak; Rauch, Isabella; Decker, Thomas; Bocklitz, Thomas; Popp, Jürgen; Gibson, Christopher M.; Fowler, Patrick W.; Huang, Wei E.; Wagner, Michael

    2015-01-01

    Microbial communities are essential to the function of virtually all ecosystems and eukaryotes, including humans. However, it is still a major challenge to identify microbial cells active under natural conditions in complex systems. In this study, we developed a new method to identify and sort active microbes on the single-cell level in complex samples using stable isotope probing with heavy water (D2O) combined with Raman microspectroscopy. Incorporation of D2O-derived D into the biomass of autotrophic and heterotrophic bacteria and archaea could be unambiguously detected via C-D signature peaks in single-cell Raman spectra, and the obtained labeling pattern was confirmed by nanoscale-resolution secondary ion MS. In fast-growing Escherichia coli cells, label detection was already possible after 20 min. For functional analyses of microbial communities, the detection of D incorporation from D2O in individual microbial cells via Raman microspectroscopy can be directly combined with FISH for the identification of active microbes. Applying this approach to mouse cecal microbiota revealed that the host-compound foragers Akkermansia muciniphila and Bacteroides acidifaciens exhibited distinctive response patterns to amendments of mucin and sugars. By Raman-based cell sorting of active (deuterated) cells with optical tweezers and subsequent multiple displacement amplification and DNA sequencing, novel cecal microbes stimulated by mucin and/or glucosamine were identified, demonstrating the potential of the nondestructive D2O-Raman approach for targeted sorting of microbial cells with defined functional properties for single-cell genomics. PMID:25550518

  4. Tracking heavy water (D2O) incorporation for identifying and sorting active microbial cells.

    PubMed

    Berry, David; Mader, Esther; Lee, Tae Kwon; Woebken, Dagmar; Wang, Yun; Zhu, Di; Palatinszky, Marton; Schintlmeister, Arno; Schmid, Markus C; Hanson, Buck T; Shterzer, Naama; Mizrahi, Itzhak; Rauch, Isabella; Decker, Thomas; Bocklitz, Thomas; Popp, Jürgen; Gibson, Christopher M; Fowler, Patrick W; Huang, Wei E; Wagner, Michael

    2015-01-13

    Microbial communities are essential to the function of virtually all ecosystems and eukaryotes, including humans. However, it is still a major challenge to identify microbial cells active under natural conditions in complex systems. In this study, we developed a new method to identify and sort active microbes on the single-cell level in complex samples using stable isotope probing with heavy water (D2O) combined with Raman microspectroscopy. Incorporation of D2O-derived D into the biomass of autotrophic and heterotrophic bacteria and archaea could be unambiguously detected via C-D signature peaks in single-cell Raman spectra, and the obtained labeling pattern was confirmed by nanoscale-resolution secondary ion MS. In fast-growing Escherichia coli cells, label detection was already possible after 20 min. For functional analyses of microbial communities, the detection of D incorporation from D2O in individual microbial cells via Raman microspectroscopy can be directly combined with FISH for the identification of active microbes. Applying this approach to mouse cecal microbiota revealed that the host-compound foragers Akkermansia muciniphila and Bacteroides acidifaciens exhibited distinctive response patterns to amendments of mucin and sugars. By Raman-based cell sorting of active (deuterated) cells with optical tweezers and subsequent multiple displacement amplification and DNA sequencing, novel cecal microbes stimulated by mucin and/or glucosamine were identified, demonstrating the potential of the nondestructive D2O-Raman approach for targeted sorting of microbial cells with defined functional properties for single-cell genomics. PMID:25550518

  5. Analytical applications of microbial fuel cells. Part II: Toxicity, microbial activity and quantification, single analyte detection and other uses.

    PubMed

    Abrevaya, Ximena C; Sacco, Natalia J; Bonetto, Maria C; Hilding-Ohlsson, Astrid; Cortón, Eduardo

    2015-01-15

    Microbial fuel cells were rediscovered twenty years ago and now are a very active research area. The reasons behind this new activity are the relatively recent discovery of electrogenic or electroactive bacteria and the vision of two important practical applications, as wastewater treatment coupled with clean energy production and power supply systems for isolated low-power sensor devices. Although some analytical applications of MFCs were proposed earlier (as biochemical oxygen demand sensing) only lately a myriad of new uses of this technology are being presented by research groups around the world, which combine both biological-microbiological and electroanalytical expertises. This is the second part of a review of MFC applications in the area of analytical sciences. In Part I a general introduction to biological-based analytical methods including bioassays, biosensors, MFCs design, operating principles, as well as, perhaps the main and earlier presented application, the use as a BOD sensor was reviewed. In Part II, other proposed uses are presented and discussed. As other microbially based analytical systems, MFCs are satisfactory systems to measure and integrate complex parameters that are difficult or impossible to measure otherwise, such as water toxicity (where the toxic effect to aquatic organisms needed to be integrated). We explore here the methods proposed to measure toxicity, microbial metabolism, and, being of special interest to space exploration, life sensors. Also, some methods with higher specificity, proposed to detect a single analyte, are presented. Different possibilities to increase selectivity and sensitivity, by using molecular biology or other modern techniques are also discussed here. PMID:24906984

  6. The effect of medium structure complexity on the growth of Saccharomyces cerevisiae in gelatin-dextran systems.

    PubMed

    Boons, Kathleen; Noriega, Estefanía; Verherstraeten, Niels; David, Charlotte C; Hofkens, Johan; Van Impe, Jan F

    2015-04-16

    As most food systems are (semi-)solid, the effect of food structure on bacterial growth has been widely acknowledged. However, studies on the growth dynamics of yeasts have neglected the effect of food structure. In this paper, the growth dynamics of the spoilage yeast Saccharomyces cerevisiae was investigated at 23.5 °C in broth, singular, homogeneous biopolymer systems and binary biopolymer systems with a heterogeneous microstructure. The biopolymers gelatin and dextran were used to introduce the different levels of structure. The metabolizing ability of gelatin and dextran by S. cerevisiae was examined. To study microbial behavior in the binary systems at the micro level, mixtures were imaged with confocal laser scanning microscopy (CLSM). Growth dynamics and microscopic images of S. cerevisiae were compared with those obtained for Escherichia coli in the same model system (Boons et al., 2014). Different phase-separated, heterogeneous microstructures were obtained by changing the amount of added gelatin and dextran. Regardless of the microstructure, S. cerevisiae was preferentially located in the dextran phase. Metabolizing ability-tests indicated that gelatin could be consumed by S. cerevisiae but in the presence of glucose, no change in gelatin concentration was observed. No indication of dextran metabolizing ability was observed. When supplementing broth with gelatin or dextran alone, an enhanced growth rate and maximum cell density were observed. This enhancement was further increased by adding a second biopolymer, introducing a heterogeneous microstructure and hence increasing the medium structure complexity. The results obtained indicate that food structure complexity plays a significant role in the growth dynamics of S. cerevisiae, an important food spoiler. PMID:25621715

  7. The Response to Heat Shock and Oxidative Stress in Saccharomyces cerevisiae

    PubMed Central

    Morano, Kevin A.; Grant, Chris M.; Moye-Rowley, W. Scott

    2012-01-01

    A common need for microbial cells is the ability to respond to potentially toxic environmental insults. Here we review the progress in understanding the response of the yeast Saccharomyces cerevisiae to two important environmental stresses: heat shock and oxidative stress. Both of these stresses are fundamental challenges that microbes of all types will experience. The study of these environmental stress responses in S. cerevisiae has illuminated many of the features now viewed as central to our understanding of eukaryotic cell biology. Transcriptional activation plays an important role in driving the multifaceted reaction to elevated temperature and levels of reactive oxygen species. Advances provided by the development of whole genome analyses have led to an appreciation of the global reorganization of gene expression and its integration between different stress regimens. While the precise nature of the signal eliciting the heat shock response remains elusive, recent progress in the understanding of induction of the oxidative stress response is summarized here. Although these stress conditions represent ancient challenges to S. cerevisiae and other microbes, much remains to be learned about the mechanisms dedicated to dealing with these environmental parameters. PMID:22209905

  8. Effect of separator and inoculum type on electricity generation and microbial community in single-chamber microbial fuel cells.

    PubMed

    Yu, Jaecheul; Park, Younghyun; Lee, Taeho

    2014-04-01

    Single-chamber microbial fuel cell (SMFC)-I consisted of 4 separator-electrode assemblies (SEAs) with two types of cation exchange membrane (CEM: Nafion and CMI 7000) and an anion exchange membrane (AEM: AMI 7001). SMFC-II consisted of 4 SEAs with Nafion and three types of nonwoven fabric. SMFC-I and -II were inoculated with anaerobic digested and activated sludge, respectively, and operated under fed-batch mode. In SMFC I, AEM-SEA showed a maximum power density (PDmax). Nafion-SEA showed a PDmax in SMFC II, which was similar to that of Nafion-SEA of SMFC I. Although different bacteria were developed in SMFC-I (Deltaproteobacteria and Firmicutes) and SMFC-II (Gammaproteobacteria, Betaproteobacteria and Bacteroidetes), the inoculum type little affects electricity generation. Variations of pH and oxygen in biofilm have influenced microbial community structure and electricity generation according to the electrode and separator material. Although the electricity generation of non-woven fabric-SEA was less than that of Nafion-SEA, the use of non-woven fabrics is expected to reduce the construction and operating costs of MFCs. PMID:24009019

  9. High power density from Pt thin film electrodes based microbial fuel cell.

    PubMed

    Sharma, Tushar; Reddy, A Leela Mohana; Chandra, T S; Ramaprabhu, S

    2008-08-01

    Microbial Fuel Cells (MFC) are robust devices capable of taping biological energy, converting sugars into potential sources of energy. Persistent efforts are directed towards increasing power output. However, they have not been researched to the extent of making them competitive with chemical fuel cells. The power generated in a dual-chamber MFC using neutral red (NR) as the electron mediator has been previously shown to be 152.4 mW/m2 at 412.5 mA/m2 of current density. In the present work we show that Pt thin film coated carbon paper as electrodes increase the performance of a microbial fuel cell compared to conventionally employed electrodes. The results obtained using E. coli based microbial fuel cell with methylene blue and neutral red as the electron mediator, potassium ferricyanide in the cathode compartment were systematically studied and the results obtained with Pt thin film coated over carbon paper as electrodes were compared with that of graphite electrodes. Platinum coated carbon electrodes were found to be better over the previously used for microbial fuel cells and at the same time are cheaper than the preferred pure platinum electrodes. PMID:19049189

  10. Copper toxicity towards Saccharomyces cerevisiae: dependence on plasma membrane fatty acid composition.

    PubMed Central

    Avery, S V; Howlett, N G; Radice, S

    1996-01-01

    One major mechanism of copper toxicity towards microorganisms is disruption of plasma membrane integrity. In this study, the influence of plasma membrane fatty acid composition on the susceptibility of Saccharomyces cerevisiae to Cu2+ toxicity was investigated. Microbial fatty acid composition is highly variable, depending on both intrinsic and environmental factors. Manipulation was achieved in this study by growth in fatty acid-supplemented medium. Whereas cells grown under standard conditions contained only saturated and monounsaturated fatty acids, considerable incorporation of the diunsaturated fatty acid linoleate (18:2) (to more than 65% of the total fatty acids) was observed in both whole-cell homogenates and plasma membrane-enriched fractions from cells grown in linoleate-supplemented medium. Linoleate enrichment had no discernible effect on the growth of S. cerevisiae. However, linoleate-enriched cells were markedly more susceptible to copper-induced plasma membrane permeabilization. Thus, after addition of Cu(NO3)2, rates of cellular K+ release (loss of membrane integrity) were at least twofold higher from linoleate-supplemented cells than from unsupplemented cells; this difference increased with reductions in the Cu2+ concentration supplied. Levels of cellular Cu accumulation were also higher in linoleate-supplemented cells. These results were correlated with a very marked dependence of whole-cell Cu2+ toxicity on cellular fatty acid unsaturation. For example, within 10 min of exposure to 5 microM Cu2+, only 3% of linoleate-enriched cells remained viable (capable of colony formation). In contrast, 100% viability was maintained in cells previously grown in the absence of a fatty acid supplement. Cells displaying intermediate levels of linoleate incorporation showed intermediate Cu2+ sensitivity, while cells enriched with the triunsaturated fatty acid linolenate (18:3) were most sensitive to Cu2+. These results demonstrate for the first time that changes

  11. Utilization of Cheese Whey Using Synergistic Immobilization of β-Galactosidase and Saccharomyces cerevisiae Cells in Dual Matrices.

    PubMed

    Kokkiligadda, Anusha; Beniwal, Arun; Saini, Priyanka; Vij, Shilpa

    2016-08-01

    Whey is a byproduct of the dairy industry, which has prospects of using as a source for production of various valuable compounds. The lactose present in whey is considered as an environmental pollutant and its utilization for enzyme and fuel production, may be effective for whey bioremediation. The dairy yeast Kluyveromyces marxianus have the ability to utilize lactose sharply as the major carbon source for the production of the enzyme. Five strains were tested for the production of the β-galactosidase using whey. The maximum β-galactosidase activity of 1.74 IU/mg dry weight was achieved in whey using K. marxianus MTCC 1389. The biocatalyst was further immobilized on chitosan macroparticles and exhibited excellent functional activity at 35 °C. Almost 89 % lactose hydrolysis was attained for concentrated whey (100 g/L) and retained 89 % catalytic activity after 15 cycles of reuse. Finally, β-galactosidase was immobilized on chitosan and Saccharomyces cerevisiae on calcium alginate, and both were used together for the production of ethanol from concentrated whey. Maximal ethanol titer of 28.9 g/L was achieved during fermentation at 35 °C. The conclusions generated by employing two different matrices will be beneficial for the future modeling using engineered S. cerevisiae in scale-up studies. PMID:27059625

  12. Single cell visualization of sulfur cycling in intertidal microbial mats

    NASA Astrophysics Data System (ADS)

    Dawson, K.; Green, A.; Orphan, V. J.

    2014-12-01

    Chemoautrophic microbial mats form in shallow intertidal pools adjacent to sulfidic hydrothermal vents in San Pedro, CA. Sulfide is primarily geologically derived. However, microscopy revealed deltaproteobacteria closely associated with Beggiatoa -like filaments, indicating an additional biogenic sulfide source, derived from sulfate reduction or sulfur disproportionation. At small scales the intercellular interaction of sulfide producing and sulfide consuming bacteria may play a important role in biogeochemical sulfur cycling. We explored the intracellular transfer of biologically derived sulfide in this system with triple and quadruple stable isotope labeling experiments: 13C, 15N, 33S, and 34S. Silicon wafers colonized by microbial mats in situ, were then incubated with 34SO42- or 34SO42- and 33S0 as well as 13C-acetate and 15NH4+and analyzed by fluorescent in situ hybridization (FISH) coupled to nanometer-scale secondary ion mass spectrometry (NanoSIMS). We observed enrichment of 34S and 33S in both deltaproteobacteria and sulfide oxidizing gammaproteobacteria. Greater enrichment relative to killed controls occurred in deltaproteobacteria than the sulfide oxidizers during both sulfate reducing (Δ34Sdelta-killed = 240‰, Δ34Sgamma-killed = 40‰) and sulfur disproportionating incubations (Δ33Sdelta-killed = 1730‰, Δ33Sgamma-killed = 1050‰). These results provide a direct visualization of interspecies sulfur transfer and indicate that biogenic sulfide derived from either sulfate or intermediate oxidation state sulfur species plays a role in sulfur cycling in this system.

  13. ChemCell : a particle-based model of protein chemistry and diffusion in microbial cells.

    SciTech Connect

    Plimpton, Steven James; Slepoy, Alexander

    2003-12-01

    Prokaryotic single-cell microbes are the simplest of all self-sufficient living organisms. Yet microbes create and use much of the molecular machinery present in more complex organisms, and the macro-molecules in microbial cells interact in regulatory, metabolic, and signaling pathways that are prototypical of the reaction networks present in all cells. We have developed a simple simulation model of a prokaryotic cell that treats proteins, protein complexes, and other organic molecules as particles which diffuse via Brownian motion and react with nearby particles in accord with chemical rate equations. The code models protein motion and chemistry within an idealized cellular geometry. It has been used to simulate several simple reaction networks and compared to more idealized models which do not include spatial effects. In this report we describe an initial version of the simulation code that was developed with FY03 funding. We discuss the motivation for the model, highlight its underlying equations, and describe simulations of a 3-stage kinase cascade and a portion of the carbon fixation pathway in the Synechococcus microbe.

  14. A hybrid microbial fuel cell stack based on single and double chamber microbial fuel cells for self-sustaining pH control

    NASA Astrophysics Data System (ADS)

    Yang, Wei; Li, Jun; Ye, Dingding; Zhang, Liang; Zhu, Xun; Liao, Qiang

    2016-02-01

    Proton accumulation in the anode chamber is the major problem that affects the operational stability and electricity generation performance of double chamber microbial fuel cells (MFCs). In this study, a hybrid microbial fuel cell stack (DS-DS stack) based on single (SCMFCs) and double chamber MFCs (DCMFCs) is proposed for self-sustaining pH control in the MFC stack. It is found that the aerobic microbial oxidation of acetate by the biofilm that is attached to the air cathode of SCMFCs is responsible for the self-sustaining removal of accumulated H+ in the effluent of DCMFCs. Compared with the stack that solely consists of SCMFCs (SS-SS stack) or DCMFCs (DD-DD stack), the hybrid stack exhibits the highest electricity output performance and the most effective conversion of acetate into electricity at high power levels. Furthermore, the hybrid stack demonstrates the operation time of 15.7 ± 1.1 h when the operating voltage is above 0.8 V. This value is much higher than that of the DD-DD (8.5 ± 2.4 h) and SS-SS (8.1 ± 1.4 h) stacks, which suggests that the hybrid stack had a good operational stability.

  15. Three Independent Forms of Regulation Affect Expression of Ho, Cln1 and Cln2 during the Cell Cycle of Saccharomyces Cerevisiae

    PubMed Central

    Breeden, L.; Mikesell, G.

    1994-01-01

    The G(1) cyclins (CLNs) bind to and activate the CDC28 kinase during the G(1) to S transition in Saccharomyces cerevisiae. Two G(1) cyclins are regulated at the RNA level so that their RNAs peak at the G(1)/S boundary. In this report we show that the cell cycle regulation of CLN1 and CLN2 is partially determined by the restricted expression of SWI4, a known trans-activator of SCB elements. When SWI4 is constitutively expressed or deleted, cell cycle regulation of CLN1/2 is reduced but not eliminated. In the absence of Swi6, another known regulator of both SCB and MCB elements, cell cycle regulation of the CLNs is also reduced, and the Start-dependence of HO transcription is eliminated. This indicates that Swi6 also plays an important role in the normal cell cycle regulation of all three promoters. When both Swi6 activity and the transcriptional regulation of SWI4 are eliminated, cell cycle regulation is further reduced, indicating that these are two independent pathways of regulation. However, a twofold fluctuation in transcript levels still persists under these conditions. This reveals a third source of cell cycle control, which could affect Swi4 activity post-transcriptionally, or reflect the existence of another unidentified regulator of these promoters. PMID:7896087

  16. Antimicrobial peptides (AMPs) produced by Saccharomyces cerevisiae induce alterations in the intracellular pH, membrane permeability and culturability of Hanseniaspora guilliermondii cells.

    PubMed

    Branco, Patrícia; Viana, Tiago; Albergaria, Helena; Arneborg, Nils

    2015-07-16

    Saccharomyces cerevisiae produces antimicrobial peptides (AMPs) during alcoholic fermentation that are active against several wine-related yeasts (e.g. Hanseniaspora guilliermondii) and bacteria (e.g. Oenococcus oeni). In the present study, the physiological changes induced by those AMPs on sensitive H. guilliermondii cells were evaluated in terms of intracellular pH (pHi), membrane permeability and culturability. Membrane permeability was evaluated by staining cells with propidium iodide (PI), pHi was determined by a fluorescence ratio imaging microscopy (FRIM) technique and culturability by a classical plating method. Results showed that the average pHi of H. guilliermondii cells dropped from 6.5 (healthy cells) to 5.4 (damaged cells) after 20 min of exposure to inhibitory concentrations of AMPs, and after 24 h 77.0% of the cells completely lost their pH gradient (∆pH=pHi-pHext). After 24h of exposure to AMPs, PI-stained (dead) cells increased from 0% to 77.7% and the number of viable cells fell from 1×10(5) to 10 CFU/ml. This means that virtually all cells (99.99%) became unculturable but that a sub-population of 22.3% of the cells remained viable (as determined by PI staining). Besides, pHi results showed that after 24h, 23% of the AMP-treated cells were sub-lethally injured (with 0<∆pH<3). Taken together, these results indicated that this subpopulation was under a viable but non-culturable (VBNC) state, which was further confirmed by recuperation assays. In summary, our study reveals that these AMPs compromise the plasma membrane integrity (and possibly also the vacuole membrane) of H. guilliermondii cells, disturbing the pHi homeostasis and inducing a loss of culturability. PMID:25897995

  17. Small ubiquitin-related modifier ligase activity of Mms21 is required for maintenance of chromosome integrity during the unperturbed mitotic cell division cycle in Saccharomyces cerevisiae.

    PubMed

    Rai, Ragini; Varma, Satya P M V; Shinde, Nikhil; Ghosh, Shilpa; Kumaran, Srikala P; Skariah, Geena; Laloraya, Shikha

    2011-04-22

    The SUMO ligase activity of Mms21/Nse2, a conserved member of the Smc5/6 complex, is required for resisting extrinsically induced genotoxic stress. We report that the Mms21 SUMO ligase activity is also required during the unchallenged mitotic cell cycle in Saccharomyces cerevisiae. SUMO ligase-defective cells were slow growing and spontaneously incurred DNA damage. These cells required caffeine-sensitive Mec1 kinase-dependent checkpoint signaling for survival even in the absence of extrinsically induced genotoxic stress. SUMO ligase-defective cells were sensitive to replication stress and displayed synthetic growth defects with DNA damage checkpoint-defective mutants such as mec1, rad9, and rad24. MMS21 SUMO ligase and mediator of replication checkpoint 1 gene (MRC1) were epistatic with respect to hydroxyurea-induced replication stress or methyl methanesulfonate-induced DNA damage sensitivity. Subjecting Mms21 SUMO ligase-deficient cells to transient replication stress resulted in enhancement of cell cycle progression defects such as mitotic delay and accumulation of hyperploid cells. Consistent with the spontaneous activation of the DNA damage checkpoint pathway observed in the Mms21-mediated sumoylation-deficient cells, enhanced frequency of chromosome breakage and loss was detected in these mutant cells. A mutation in the conserved cysteine 221 that is engaged in coordination of the zinc ion in Loop 2 of the Mms21 SPL-RING E3 ligase catalytic domain resulted in strong replication stress sensitivity and also conferred slow growth and Mec1 dependence to unchallenged mitotically dividing cells. Our findings establish Mms21-mediated sumoylation as a determinant of cell cycle progression and maintenance of chromosome integrity during the unperturbed mitotic cell division cycle in budding yeast. PMID:21324902

  18. Use of a Burkholderia cenocepacia ABTS Oxidizer in a Microbial Fuel Cell

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Microbial fuel cells (MFCs) often use biological processes to generate electrons from organic material contained in the anode chamber and abiotic processes employing atmospheric oxygen as the oxidant in the cathode chamber. This study investigated the accumulation of an oxidant in bacterial cultures...

  19. Microbial electrodialysis cell for simultaneous water desalination and hydrogen gas production.

    PubMed

    Mehanna, Maha; Kiely, Patrick D; Call, Douglas F; Logan, Bruce E

    2010-12-15

    A new approach to water desalination is to use exoelectrogenic bacteria to generate electrical power from the biodegradation of organic matter, moving charged ions from a middle chamber between two membranes in a type of microbial fuel cell called a microbial desalination cell. Desalination efficiency using this approach is limited by the voltage produced by the bacteria. Here we examine an alternative strategy based on boosting the voltage produced by the bacteria to achieve hydrogen gas evolution from the cathode using a three-chambered system we refer to as a microbial electrodialysis cell (MEDC). We examined the use of the MEDC process using two different initial NaCl concentrations of 5 g/L and 20 g/L. Conductivity in the desalination chamber was reduced by up to 68 ± 3% in a single fed-batch cycle, with electrical energy efficiencies reaching 231 ± 59%, and maximum hydrogen production rates of 0.16 ± 0.05 m(3) H(2)/m(3) d obtained at an applied voltage of 0.55 V. The advantage of this system compared to a microbial fuel cell approach is that the potentials between the electrodes can be better controlled, and the hydrogen gas that is produced can be used to recover energy to make the desalination process self-sustaining with respect to electrical power requirements. PMID:21077623

  20. Influence of different buffers (HEPES/MOPS) on keratinocyte cell viability and microbial growth.

    PubMed

    Dias, Kássia de Carvalho; Barbugli, Paula Aboud; Vergani, Carlos Eduardo

    2016-06-01

    This study assessed the effect of the buffers 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) and 3-(N-morpholino) propanesulfonic acid (MOPS) on keratinocyte cell viability and microbial growth. It was observed that RPMI buffered with HEPES, supplemented with l-glutamine and sodium bicarbonate, can be used as a more suitable medium to promote co-culture. PMID:27060444

  1. Triclosan Alters Anti-microbial and Inflammatory Responses of Epithelial Cells

    PubMed Central

    Wallet, Mark A.; Calderon, Nadia L.; Alonso, Tess R.; Choe, Christina S.; Catalfamo, Dana L.; Lalane, Charles J.; Neiva, Kathleen G.; Panagakos, Foti; Wallet, Shannon M.

    2012-01-01

    Periodontal diseases are a class of pathologies wherein oral microbes induce harmful immune responses in a susceptible host. Therefore, an agent which can both reduce microbial burden and lessen pathogenesis of localized inflammation would have beneficial effects in periodontal disease. 2,4,4-trichloro-2-hydroxydiphenyl-ether [triclosan] is currently used in oral care products due to broad spectrum anti-microbial and anti-inflammatory properties. Objective To determine effects of triclosan on the response of oral epithelial cells to stimulation with the inflammatory microbial product lipopolysaccharide [LPS], a ligand for toll-like receptor 4 [TLR4]. Materials/Methods Primary human oral epithelial cells were stimulated with LPS in the presence and/or absence of triclosan after which expression of pro-inflammatory cytokines, β-defensins, micro-RNAs [miRNAs] or TLR signaling pathway proteins were evaluated. Results Here we demonstrate that triclosan is a potent inhibitor of oral epithelial cell LPS-induced pro-inflammatory responses by inducing miRNA regulation of the TLR-signaling pathway. Triclosan was not a pan-suppresser of oral epithelial cell responses as β-defensin 2 [βD2] and βD3 were upregulated by triclosan following LPS-stimulation. Conclusions These data demonstrate both a novel anti-microbial mechanism by which triclosan improves plaque control and an additional anti-inflammatory property which could have beneficial effects in periodontal disease resolution. PMID:24079913

  2. Utilizing the green alga Chlamydomonas reinhardtii for microbial electricity generation: a living solar cell.

    PubMed

    Rosenbaum, Miriam; Schröder, Uwe; Scholz, Fritz

    2005-10-01

    By employing living cells of the green alga Chlamydomonas reinhardtii, we demonstrate the possibility of direct electricity generation from microbial photosynthetic activity. The presented concept is based on an in situ oxidative depletion of hydrogen, photosynthetically produced by C. reinhardtii under sulfur-deprived conditions, by polymer-coated electrocatalytic electrodes. PMID:15696280

  3. Synthesization of SnO2-modified carbon nanotubes and their application in microbial fuel cell

    NASA Astrophysics Data System (ADS)

    Wang, Zi-Bo; Xiong, Shi-Chang; Guan, Yu-Jiang; Zhu, Xue-Qiang

    2016-03-01

    The aim of this work was to study the synthesization of SnO2-modified carbon nanotubes and their application in microbial fuel cell. With the chemical vapor deposition technique, carbon nanotubes growing in situ on a carbon felt are obtained. A SnO2 sol was applied to the carbon felt to prepare a SnO2-modified carbon nanotubes. X-ray diffraction and energy-dispersive X-ray analysis confirmed that SnO2 existed in the prepared samples. Using the prepared samples as anode electrodes, flexible graphite as cathode, and glucose solution as substrate in microbial fuel cell, the effects of the temperature, substrate concentration, and electrodes on removal rates for chemical oxygen demand and the performance of microbial fuel cell have been analyzed. With substrate concentration of 1500 mg L-1, the microbial fuel cell had an optimal output voltage of 563 mV and a removal rate of 78 % for chemical oxygen demand at 311 K. The composite electrodes are stable and reusable.

  4. Glycan-specific whole cell affinity chromatography: a versatile microbial adhesion platform

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We have constructed a C-glycoside ketohydrazide affinity chromatography resin that interacts with viable whole-cell microbial populations with biologically appropriate stereo-specificity in a carbohydrate-defined manner. It readily allows for the quantification, selection, and manipulation of target...

  5. Cadmium biosorption by Saccharomyces cerevisiae

    SciTech Connect

    Volesky, B.; May, H.; Holan, Z.R. )

    1993-04-01

    Cadmium uptake by nonliving and resting cells of Saccharomyces cerevisiae obtained from aerobic or anaerobic cultures from pure cadmium-bearing solutions was examined. The highest cadmium uptake exceeding 70 mg Cd/g was observed with aerobic baker's yeast biomass from the exponential growth phase. Nearly linear sorption isotherms featured by higher sorbing resting cells together with metal deposits localized exclusively in vacuoles indicate the possibility of a different metal-sequestering mechanism when compared to dry nonliving yeasts which did not usually accumulate more than 20 mg Cd/g. The uptake of cadmium was relatively fast, 75% of the sorption completed in less than 5 min.

  6. Transcription activator-like effector nucleases mediated metabolic engineering for enhanced fatty acids production in Saccharomyces cerevisiae.

    PubMed

    Aouida, Mustapha; Li, Lixin; Mahjoub, Ali; Alshareef, Sahar; Ali, Zahir; Piatek, Agnieszka; Mahfouz, Magdy M

    2015-10-01

    Targeted engineering of microbial genomes holds much promise for diverse biotechnological applications. Transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats/Cas9 systems are capable of efficiently editing microbial genomes, including that of Saccharomyces cerevisiae. Here, we demonstrate the use of TALENs to edit the genome of S. cerevisiae with the aim of inducing the overproduction of fatty acids. Heterodimeric TALENs were designed to simultaneously edit the FAA1 and FAA4 genes encoding acyl-CoA synthetases in S. cerevisiae. Functional yeast double knockouts generated using these TALENs over-produce large amounts of free fatty acids into the cell. This study demonstrates the use of TALENs for targeted engineering of yeast and demonstrates that this technology can be used to stimulate the enhanced production of free fatty acids, which are potential substrates for biofuel production. This proof-of-principle study extends the utility of TALENs as excellent genome editing tools and highlights their potential use for metabolic engineering of yeast and other organisms, such as microalgae and plants, for biofuel production. PMID:25907574

  7. Prospects of microbial cell factories developed through systems metabolic engineering.

    PubMed

    Gustavsson, Martin; Lee, Sang Yup

    2016-09-01

    While academic-level studies on metabolic engineering of microorganisms for production of chemicals and fuels are ever growing, a significantly lower number of such production processes have reached commercial-scale. In this work, we review the challenges associated with moving from laboratory-scale demonstration of microbial chemical or fuel production to actual commercialization, focusing on key requirements on the production organism that need to be considered during the metabolic engineering process. Metabolic engineering strategies should take into account techno-economic factors such as the choice of feedstock, the product yield, productivity and titre, and the cost effectiveness of midstream and downstream processes. Also, it is important to develop an industrial strain through metabolic engineering for pathway construction and flux optimization together with increasing tolerance to products and inhibitors present in the feedstock, and ensuring genetic stability and strain robustness under actual fermentation conditions. PMID:27435545

  8. Nanomechanical sensors for single microbial cell growth monitoring

    NASA Astrophysics Data System (ADS)

    Maloney, Niall; Lukacs, Gyongyi; Jensen, Jason; Hegner, Martin

    2014-06-01

    A nanomechanical technique for rapid real time detection and monitoring of microorganism growth will significantly reduce costs and diagnosis times in industrial and clinical settings. Owing to their label free detection mechanism and unprecedented sensitivity to the mass and elastic modulus of biological structures, dynamically operated cantilever arrays provide an opportunity to rapidly detect and track the evolution of microbial growth. Here we report the monitoring of the growth of single Aspergillus niger spores via the multimode response of microcantilevers. The fungal hyphal structure affects the cantilevers' nanomechanical properties as it propagates along the sensor. We demonstrate, for the first time, the mapping of cellular events with great accuracy using a cantilever frequency response. Imaging of growth conditions on the cantilever, which is performed in parallel, allows for verification of these results. Theoretical comparison and finite element modelling confirm experimental findings and allow for determination of the hyphal elastic modulus.A nanomechanical technique for rapid real time detection and monitoring of microorganism growth will significantly reduce costs and diagnosis times in industrial and clinical settings. Owing to their label free detection mechanism and unprecedented sensitivity to the mass and elastic modulus of biological structures, dynamically operated cantilever arrays provide an opportunity to rapidly detect and track the evolution of microbial growth. Here we report the monitoring of the growth of single Aspergillus niger spores via the multimode response of microcantilevers. The fungal hyphal structure affects the cantilevers' nanomechanical properties as it propagates along the sensor. We demonstrate, for the first time, the mapping of cellular events with great accuracy using a cantilever frequency response. Imaging of growth conditions on the cantilever, which is performed in parallel, allows for verification of these

  9. Rapid prototyping of microbial cell factories via genome-scale engineering.

    PubMed

    Si, Tong; Xiao, Han; Zhao, Huimin

    2015-11-15

    Advances in reading, writing and editing genetic materials have greatly expanded our ability to reprogram biological systems at the resolution of a single nucleotide and on the scale of a whole genome. Such capacity has greatly accelerated the cycles of design, build and test to engineer microbes for efficient synthesis of fuels, chemicals and drugs. In this review, we summarize the emerging technologies that have been applied, or are potentially useful for genome-scale engineering in microbial systems. We will focus on the development of high-throughput methodologies, which may accelerate the prototyping of microbial cell factories. PMID:25450192

  10. Engineering PQS Biosynthesis Pathway for Enhancement of Bioelectricity Production in Pseudomonas aeruginosa Microbial Fuel Cells

    PubMed Central

    Cao, Bin; Seviour, Thomas; Nesatyy, Victor J.; Marsili, Enrico; Kjelleberg, Staffan; Givskov, Michael; Tolker-Nielsen, Tim; Song, Hao; Loo, Joachim Say Chye; Yang, Liang

    2013-01-01

    The biosynthesis of the redox shuttle, phenazines, in Pseudomonas aeruginosa, an ubiquitous microorganism in wastewater microflora, is regulated by the 2-heptyl-3,4-dihydroxyquinoline (PQS) quorum-sensing system. However, PQS inhibits anaerobic growth of P. aeruginosa. We constructed a P. aeruginosa strain that produces higher concentrations of phenazines under anaerobic conditions by over-expressing the PqsE effector in a PQS negative ΔpqsC mutant. The engineered strain exhibited an improved electrical performance in microbial fuel cells (MFCs) and potentiostat-controlled electrochemical cells with an approximate five-fold increase of maximum current density relative to the parent strain. Electrochemical analysis showed that the current increase correlates with an over-synthesis of phenazines. These results therefore demonstrate that targeting microbial cell-to-cell communication by genetic engineering is a suitable technique to improve power output of bioelectrochemical systems. PMID:23700414

  11. Coating-type three-dimensional acetate-driven microbial fuel cells.

    PubMed

    Yu, Jin; Tang, Yulan

    2015-08-01

    This study uses sodium acetate as fuel to construct bioelectricity in coating-type three-dimensional microbial fuel cells anode. The coating-type three-dimensional anode was constructed using iron net as structural support, adhering a layer of carbon felt as primary coating and using carbon powder and 30% PTFE solution mixture as coating. The efficiency of electricity production and wastewater treatment were analyzed for the three-dimensional acetate-fed (C2H3NaO2) microbial fuel cells with the various ratio of the coating mixture. The results showed that the efficiency of electricity production was significantly improved when using the homemade coating-type microbial fuel cells anode compared with the one without coating on the iron net, which the apparent internal resistance was decreased by 59.4% and the maximum power density was increased by 1.5 times. It was found the electricity production was greatly influenced by the ratio of the carbon powder and PTFE in the coating. The electricity production was the highest with apparent internal resistance of 190 Ω, and maximum power density of 5189.4 mW m(-3) when 750 mg of carbon powder and 10 ml of PTFE (i.e., ratio 75:1) was used in the coating. With the efficiency of electricity production, wide distribution and low cost of the raw materials, the homemade acetate-fed microbial fuel cells provides a valuable reference to the development of the composition microbial fuel cell anode production. PMID:25681073

  12. Cleaning-up atrazine-polluted soil by using Microbial Electroremediating Cells.

    PubMed

    Domínguez-Garay, Ainara; Boltes, Karina; Esteve-Núñez, Abraham

    2016-10-01

    Biodegradation of pollutants in soil is greatly limited by the availability of terminal electron acceptors required for supporting microbial respiration. Such limitation can be overcome if soil-buried electrodes accept the electrons released in the microbial metabolism. We propose the term bioelectroventing for such a environmental treatment. The process would be performed in a device so-called Microbial Electroremediating Cell. Indeed, our studies demonstrate that the presence of electrodes as electron acceptors effectively stimulated by 5-fold the biodegradation rate of the herbicide atrazine (2-chloro-4-ethylamino-6-isopropyl amino-1,3,5-triazine) in comparison with soil natural attenuation. Furthermore, a different set of toxicological test using Pseudokirchneriella subcapitata green alga e, Salmonella typhimorium bacteria and Sorghum saccharatum plant seeds respectively, confirm that atrazine-polluted soil can be effectively cleaned-up in short time by the use of MERCs. PMID:27448317

  13. The Role of Synthetic Biology in the Design of Microbial Cell Factories for Biofuel Production

    PubMed Central

    Colin, Verónica Leticia; Rodríguez, Analía; Cristóbal, Héctor Antonio

    2011-01-01

    Insecurity in the supply of fossil fuels, volatile fuel prices, and major concerns regarding climate change have sparked renewed interest in the production of fuels from renewable resources. Because of this, the use of biodiesel has grown dramatically during the last few years and is expected to increase even further in the future. Biodiesel production through the use of microbial systems has marked a turning point in the field of biofuels since it is emerging as an attractive alternative to conventional technology. Recent progress in synthetic biology has accelerated the ability to analyze, construct, and/or redesign microbial metabolic pathways with unprecedented precision, in order to permit biofuel production that is amenable to industrial applications. The review presented here focuses specifically on the role of synthetic biology in the design of microbial cell factories for efficient production of biodiesel. PMID:22028591

  14. Microbial production of value-added nutraceuticals.

    PubMed

    Wang, Jian; Guleria, Sanjay; Koffas, Mattheos A G; Yan, Yajun

    2016-02-01

    Nutraceuticals are important natural bioactive compounds that confer health-promoting and medical benefits to humans. Globally growing demands for value-added nutraceuticals for prevention and treatment of human diseases have rendered nutraceuticals a multi-billion dollar market. However, supply limitations and extraction difficulties from natural sources such as plants, animals or fungi, restrict the large-scale use of nutraceuticals. Metabolic engineering via microbial production platforms has been advanced as an eco-friendly alternative approach for production of value-added nutraceuticals from simple carbon sources. Microbial platforms like the most widely used Escherichia coli and Saccharomyces cerevisiae have been engineered as versatile cell factories for production of diverse and complex value-added chemicals such as phytochemicals, prebiotics, polysaccaharides and poly amino acids. This review highlights the recent progresses in biological production of value-added nutraceuticals via metabolic engineering approaches. PMID:26716360

  15. Rapid resonance Raman microspectroscopy to probe carbon dioxide fixation by single cells in microbial communities

    PubMed Central

    Li, Mengqiu; Canniffe, Daniel P; Jackson, Philip J; Davison, Paul A; FitzGerald, Simon; Dickman, Mark J; Burgess, J Grant; Hunter, C Neil; Huang, Wei E

    2012-01-01

    Photosynthetic microorganisms play crucial roles in aquatic ecosystems and are the major primary producers in global marine ecosystems. The discovery of new bacteria and microalgae that play key roles in CO2 fixation is hampered by the lack of methods to identify hitherto-unculturable microorganisms. To overcome this problem we studied single microbial cells using stable-isotope probing (SIP) together with resonance Raman (RR) microspectroscopy of carotenoids, the light-absorbing pigments present in most photosynthetic microorganisms. We show that fixation of 13CO2 into carotenoids produces a red shift in single-cell RR (SCRR) spectra and that this SCRR–SIP technique is sufficiently sensitive to detect as little as 10% of 13C incorporation. Mass spectrometry (MS) analysis of labelled cellular proteins verifies that the red shift in carotenoid SCRR spectra acts as a reporter of the 13C content of single cells. Millisecond Raman imaging of cells in mixed cultures and natural seawater samples was used to identify cells actively fixing CO2, demonstrating that the SCRR–SIP is a noninvasive method for the rapid and quantitative detection of CO2 fixation at the single cell level in a microbial community. The SCRR–SIP technique may provide a direct method for screening environmental samples, and could help to reveal the ecophysiology of hitherto-unculturable microorganisms, linking microbial species to their ecological function in the natural environment. PMID:22113377

  16. PKS and NRPS gene clusters from microbial symbiont cells of marine sponges by whole genome amplification.

    PubMed

    Siegl, Alexander; Hentschel, Ute

    2010-08-01

    Whole genome amplification (WGA) approaches provide genomic information on single microbial cells and hold great promise for the field of environmental microbiology. Here, the microbial consortia of the marine sponge Aplysina aerophoba were sorted by fluorescence-activated cell sorting (FACS) and then subjected to WGA. A cosmid library was constructed from the WGA product of a sample containing two bacterial cells, one a member of the candidate phylum Poribacteria and one of a sponge-specific clade of Chloroflexi. Library screening led to the genomic characterization of three cosmid clones, encoding a polyketide synthase (PKS), a non-ribosomal peptide synthetase (NRPS) and the Chloroflexi 16S rRNA gene. PCR screening of WGA products from additional, FACS-sorted single bacterial symbiont cells supports the assignment of the Sup-PKS gene to the Poribacteria and the novel NRPS gene to the Chloroflexi. This promising single-cell genomics approach has permitted cloning of entire gene clusters from single microbial cells of known phylogenetic origin and thus provides a sought-after link between phylogeny and function. PMID:23766222

  17. Composition of EPS fractions from suspended sludge and biofilm and their roles in microbial cell aggregation.

    PubMed

    Zhang, Peng; Fang, Fang; Chen, You-Peng; Shen, Yu; Zhang, Wei; Yang, Ji-Xiang; Li, Chun; Guo, Jin-Song; Liu, Shao-Yang; Huang, Yang; Li, Shan; Gao, Xu; Yan, Peng

    2014-12-01

    The adhesion and aggregation properties of microbial cell are closely related to extracellular polymeric substances (EPS). In this work, the composition and physicochemical characteristics of EPS in biofilm and suspended sludge (S-sludge) were determined to evaluate their roles in microbial cell aggregation. Raman spectroscopy and three-dimensional fluorescence spectra have been employed to reveal each EPS fraction in different composition. The flocculating capacity of each EPS fraction in the S-sludge shows extraordinary activity, comparing its counterpart in biofilm. Microbial cell surfaces present high hydrophobicity and increased zeta potentials upon EPS extraction. In addition, the respective contribution of EPS to cell aggregating was elucidated. The contribution of combined SEPS and LB-EPS was 23% for S-sludge sample, whereas that was negligible for biofilm sample. The contribution of LB-EPS and TB-EPS were 16% and 30% for S-sludge sample, and -6% and negligible for biofilm sample, respectively. Therefore, EPS promoted the S-sludge cells to aggregate, while in contrast, they showed a negligible or negative effect on the biofilm cells aggregating. PMID:24968163

  18. GroE chaperonins assisted functional expression of bacterial enzymes in Saccharomyces cerevisiae.

    PubMed

    Xia, Peng-Fei; Zhang, Guo-Chang; Liu, Jing-Jing; Kwak, Suryang; Tsai, Ching-Sung; Kong, In Iok; Sung, Bong Hyun; Sohn, Jung-Hoon; Wang, Shu-Guang; Jin, Yong-Su

    2016-10-01

    Rapid advances in the capabilities of reading and writing DNA along with increasing understanding of microbial metabolism at the systems-level have paved an incredible path for metabolic engineering. Despite these advances, post-translational tools facilitating functional expression of heterologous enzymes in model hosts have not been developed well. Some bacterial enzymes, such as Escherichia coli xylose isomerase (XI) and arabinose isomerase (AI) which are essential for utilizing cellulosic sugars, cannot be functionally expressed in Saccharomyces cerevisiae. We hypothesized and demonstrated that the mismatching of the HSP60 chaperone systems between bacterial and eukaryotic cells might be the reason these bacterial enzymes cannot be functionally expressed in yeast. The results showed that the co-expression of E. coli GroE can facilitate the functional expression of E. coli XI and AI, as well as the Agrobacterium tumefaciens D-psicose epimerase in S. cerevisiae. The co-expression of bacterial chaperonins in S. cerevisiae is a promising post-translational strategy for the functional expression of bacterial enzymes in yeast. Biotechnol. Bioeng. 2016;113: 2149-2155. © 2016 Wiley Periodicals, Inc. PMID:27003667

  19. Metabolic engineering of Saccharomyces cerevisiae for production of fatty acid-derived biofuels and chemicals.

    PubMed

    Runguphan, Weerawat; Keasling, Jay D

    2014-01-01

    As the serious effects of global climate change become apparent and access to fossil fuels becomes more limited, metabolic engineers and synthetic biologists are looking towards greener sources for transportation fuels. In recent years, microbial production of high-energy fuels by economically efficient bioprocesses has emerged as an attractive alternative to the traditional production of transportation fuels. Here, we engineered the budding yeast Saccharomyces cerevisiae to produce fatty acid-derived biofuels and chemicals from simple sugars. Specifically, we overexpressed all three fatty acid biosynthesis genes, namely acetyl-CoA carboxylase (ACC1), fatty acid synthase 1 (FAS1) and fatty acid synthase 2 (FAS2), in S. cerevisiae. When coupled to triacylglycerol (TAG) production, the engineered strain accumulated lipid to more than 17% of its dry cell weight, a four-fold improvement over the control strain. Understanding that TAG cannot be used directly as fuels, we also engineered S. cerevisiae to produce drop-in fuels and chemicals. Altering the terminal "converting enzyme" in the engineered strain led to the production of free fatty acids at a titer of approximately 400 mg/L, fatty alcohols at approximately 100mg/L and fatty acid ethyl esters (biodiesel) at approximately 5 mg/L directly from simple sugars. We envision that our approach will provide a scalable, controllable and economic route to this important class of chemicals. PMID:23899824

  20. A microfluidic microbial fuel cell array that supports long-term multiplexed analyses of electricigens.

    PubMed

    Hou, Huijie; Li, Lei; Ceylan, Cemile Ümran; Haynes, Abria; Cope, Julia; Wilkinson, Heather H; Erbay, Celal; de Figueiredo, Paul; Han, Arum

    2012-10-21

    Microbial fuel cells (MFCs) are green energy technologies that exploit microbial metabolism to generate electricity. The widespread implementation of MFC technologies has been stymied by their high cost and limited power. MFC arrays in which device configurations or microbial consortia can be screened have generated significant interest because of their potential for defining aspects that will improve performance featuring high throughput characteristics. However, current miniature MFCs and MFC array systems do not support long-term studies that mimic field conditions, and hence, have limitations in fully characterizing and understanding MFC performances in varieties of conditions. Here, we describe an MFC array device that incorporates microfluidic technology to enable continuous long-term analysis of MFC performance at high throughput utilizing periodic anolyte/catholyte replenishment. The system showed 360% higher power output and 700% longer operating time when compared to MFC arrays without catholyte replenishment. We further demonstrate the utility of the system by reporting its successful use in screening microbial consortia collected from geographically diverse environments for communities that support enhanced MFC performance. Taken together, this work demonstrates that anolyte/catholyte replenishment can significantly improve the long-term performance of microfabricated MFC arrays, and support the characterization of diverse microbial consortia. PMID:22868338

  1. Microbial response to single-cell protein production and brewery wastewater treatment

    PubMed Central

    Lee, Jackson Z; Logan, Andrew; Terry, Seth; Spear, John R

    2015-01-01

    As global fisheries decline, microbial single-cell protein (SCP) produced from brewery process water has been highlighted as a potential source of protein for sustainable animal feed. However, biotechnological investigation of SCP is difficult because of the natural variation and complexity of microbial ecology in wastewater bioreactors. In this study, we investigate microbial response across a full-scale brewery wastewater treatment plant and a parallel pilot bioreactor modified to produce an SCP product. A pyrosequencing survey of the brewery treatment plant showed that each unit process selected for a unique microbial community. Notably, flow equalization basins were dominated by Prevotella, methanogenesis effluent had the highest levels of diversity, and clarifier wet-well samples were sources of sequences for the candidate bacterial phyla of TM7 and BD1-5. Next, the microbial response of a pilot bioreactor producing SCP was tracked over 1 year, showing that two different production trials produced two different communities originating from the same starting influent. However, SCP production resulted generally in enrichment of several clades of rhizospheric diazotrophs of Alphaproteobacteria and Betaproteobacteria in the bioreactor and even more so in the final product. These diazotrophs are potentially useful as the basis of a SCP product for commercial feed production. PMID:24837420

  2. Microbial response to single-cell protein production and brewery wastewater treatment.

    PubMed

    Lee, Jackson Z; Logan, Andrew; Terry, Seth; Spear, John R

    2015-01-01

    As global fisheries decline, microbial single-cell protein (SCP) produced from brewery process water has been highlighted as a potential source of protein for sustainable animal feed. However, biotechnological investigation of SCP is difficult because of the natural variation and complexity of microbial ecology in wastewater bioreactors. In this study, we investigate microbial response across a full-scale brewery wastewater treatment plant and a parallel pilot bioreactor modified to produce an SCP product. A pyrosequencing survey of the brewery treatment plant showed that each unit process selected for a unique microbial community. Notably, flow equalization basins were dominated by Prevotella, methanogenesis effluent had the highest levels of diversity, and clarifier wet-well samples were sources of sequences for the candidate bacterial phyla of TM7 and BD1-5. Next, the microbial response of a pilot bioreactor producing SCP was tracked over 1 year, showing that two different production trials produced two different communities originating from the same starting influent. However, SCP production resulted generally in enrichment of several clades of rhizospheric diazotrophs of Alphaproteobacteria and Betaproteobacteria in the bioreactor and even more so in the final product. These diazotrophs are potentially useful as the basis of a SCP product for commercial feed production. PMID:24837420

  3. Effects of proton exchange membrane on the performance and microbial community composition of air-cathode microbial fuel cells.

    PubMed

    Lee, Yun-Yeong; Kim, Tae Gwan; Cho, Kyung-Suk

    2015-10-10

    This study investigated the effects of proton exchange membranes (PEMs) on performance and microbial community of air-cathode microbial fuel cells (MFCs). Air-cathode MFCs with reactor volume of 1L were constructed in duplicate with or without PEM (designated as ACM-MFC and AC-MFC, respectively) and fed with a mixture of glucose and acetate (1:1, w:w). The maximum power density and coulombic efficiency did not differ between MFCs in the absence or presence of a PEM. However, PEM use adversely affected maximum voltage production and the rate of organic compound removal (p<0.05). Quantitative droplet digital PCR indicated that AC-MFCs had a greater bacterial population than ACM-MFCs (p<0.05). Likewise, ribosomal tag pyrosequencing revealed that the diversity index of bacterial communities was greater for AC-MFCs (p<0.05). Network analysis revealed that the most abundant genus was Enterococcus, which comprised ≥62% of the community and was positively associated with PEM and negatively associated with the rate of chemical oxygen demand (COD) removal (Pearson correlation>0.9 and p<0.05). Geobacter, which is known as an exoelectrogen, was positively associated with maximum power density and negatively associated with PEM. Thus, these results suggest that the absence of PEM favored the growth of Geobacter, a key player for electricity generation in MFC systems. Taken together, these findings demonstrate that MFC systems without PEM are more efficient with respect to power production and COD removal as well as exoelectrogen growth. PMID:26235818

  4. Response of Saccharomyces cerevisiae to cadmium and nickel stress: the use of the sugar cane vinasse as a potential mitigator.

    PubMed

    Oliveira, Ricardo Pinheiro de Souza; Basso, Luiz Carlos; Junior, Adalberto Pessoa; Penna, Thereza Christina Vessoni; Del Borghi, Marco; Converti, Attilio

    2012-01-01

    Most of the metals released from industrial activity, among them are cadmium (Cd) and nickel (Ni), inhibit the productivity of cultures and affect microbial metabolism. In this context, the aim of this work was to investigate the capacity of sugar cane vinasse to mitigate the adverse effects of Cd and Ni on cell growth, viability, budding rate and trehalose content of Saccharomyces cerevisiae, likely because of adsorption and chelating action. For this purpose, the yeast was grown batch-wise in YED medium supplemented with selected amounts of vinasse and Cd or Ni. The negative effects of Cd and Ni on S. cerevisiae growth and the mitigating one of sugar cane vinasse were quantified by an exponential model. Without vinasse, the addition of increasing levels of Cd and Ni reduced the specific growth rate, whereas in its presence no reduction was observed. Consistently with the well-proved toxicity of both metals, cell viability and budding rate progressively decreased with increasing their concentration, but in the presence of vinasse the situation was remarkably improved. The trehalose content of S. cerevisiae cells followed the same qualitative behavior as cell viability, even though the negative effect of both metals on this parameter was stronger. These results demonstrate the ability of sugar cane vinasse to mitigate the toxic effects of Cd and Ni. PMID:21809054

  5. A Genomic Screen Revealing the Importance of Vesicular Trafficking Pathways in Genome Maintenance and Protection against Genotoxic Stress in Diploid Saccharomyces cerevisiae Cells

    PubMed Central

    Krol, Kamil; Brozda, Izabela; Skoneczny, Marek; Bretne, Maria; Skoneczna, Adrianna

    2015-01-01

    The ability to survive stressful conditions is important for every living cell. Certain stresses not only affect the current well-being of cells but may also have far-reaching consequences. Uncurbed oxidative stress can cause DNA damage and decrease cell survival and/or increase mutation rates, and certain substances that generate oxidative damage in the cell mainly act on DNA. Radiomimetic zeocin causes oxidative damage in DNA, predominantly by inducing single- or double-strand breaks. Such lesions can lead to chromosomal rearrangements, especially in diploid cells, in which the two sets of chromosomes facilitate excessive and deleterious recombination. In a global screen for zeocin-oversensitive mutants, we selected 133 genes whose deletion reduces the survival of zeocin-treated diploid Saccharomyces cerevisiae cells. The screen revealed numerous genes associated with stress responses, DNA repair genes, cell cycle progression genes, and chromatin remodeling genes. Notably, the screen also demonstrated the involvement of the vesicular trafficking system in cellular protection against DNA damage. The analyses indicated the importance of vesicular system integrity in various pathways of cellular protection from zeocin-dependent damage, including detoxification and a direct or transitional role in genome maintenance processes that remains unclear. The data showed that deleting genes involved in vesicular trafficking may lead to Rad52 focus accumulation and changes in total DNA content or even cell ploidy alterations, and such deletions may preclude proper DNA repair after zeocin treatment. We postulate that functional vesicular transport is crucial for sustaining an integral genome. We believe that the identification of numerous new genes implicated in genome restoration after genotoxic oxidative stress combined with the detected link between vesicular trafficking and genome integrity will reveal novel molecular processes involved in genome stability in diploid cells

  6. Transporter engineering for improved tolerance against alkane biofuels in Saccharomyces cerevisiae

    PubMed Central

    2013-01-01

    Background Hydrocarbon alkanes, components of major fossil fuels, are considered as next-generation biofuels because their biological production has recently been shown to be possible. However, high-yield alkane production requires robust host cells that are tolerant against alkanes, which exhibit cytotoxicity. In this study, we aimed to improve alkane tolerance in Saccharomyces cerevisiae, a key industrial microbial host, by harnessing heterologous transporters that potentially pump out alkanes. Results To this end, we attempted to exploit ABC transporters in Yarrowia lipolytica based on the observation that it utilizes alkanes as a carbon source. We confirmed the increased transcription of ABC2 and ABC3 transporters upon exposure to a range of alkanes in Y. lipolytica. We then showed that the heterologous expression of ABC2 and ABC3 transporters significantly increased tolerance against decane and undecane in S. cerevisiae through maintaining lower intracellular alkane level. In particular, ABC2 transporter increased the tolerance limit of S. cerevisiae about 80-fold against decane. Furthermore, through site-directed mutagenesis for glutamate (E988 for ABC2, and E989 for ABC3) and histidine (H1020 for ABC2, and H1021 for ABC3), we provided the evidence that glutamate was essential for the activity of ABC2 and ABC3 transporters, with ATP most likely to be hydrolyzed by a catalytic carboxylate mechanism. Conclusions Here, we demonstrated that transporter engineering through expression of heterologous efflux pumps led to significantly improved tolerance against alkane biofuels in S. cerevisiae. We believe that our results laid the groundwork for developing robust alkane-producing yeast cells through transporter engineering, which will greatly aid in next-generation alkane biofuel production and recovery. PMID:23402697

  7. The Morphogenesis Checkpoint in Saccharomyces cerevisiae: Cell Cycle Control of Swe1p Degradation by Hsl1p and Hsl7p

    PubMed Central

    McMillan, John N.; Longtine, Mark S.; Sia, Rey A. L.; Theesfeld, Chandra L.; Bardes, Elaine S. G.; Pringle, John R.; Lew, Daniel J.

    1999-01-01

    In Saccharomyces cerevisiae, the Wee1 family kinase Swe1p is normally stable during G1 and S phases but is unstable during G2 and M phases due to ubiquitination and subsequent degradation. However, perturbations of the actin cytoskeleton lead to a stabilization and accumulation of Swe1p. This response constitutes part of a morphogenesis checkpoint that couples cell cycle progression to proper bud formation, but the basis for the regulation of Swe1p degradation by the morphogenesis checkpoint remains unknown. Previous studies have identified a protein kinase, Hsl1p, and a phylogenetically conserved protein of unknown function, Hsl7p, as putative negative regulators of Swe1p. We report here that Hsl1p and Hsl7p act in concert to target Swe1p for degradation. Both proteins are required for Swe1p degradation during the unperturbed cell cycle, and excess Hsl1p accelerates Swe1p degradation in the G2-M phase. Hsl1p accumulates periodically during the cell cycle and promotes the periodic phosphorylation of Hsl7p. Hsl7p can be detected in a complex with Swe1p in cell lysates, and the overexpression of Hsl7p or Hsl1p produces an effective override of the G2 arrest imposed by the morphogenesis checkpoint. These findings suggest that Hsl1p and Hsl7p interact directly with Swe1p to promote its recognition by the ubiquitination complex, leading ultimately to its destruction. PMID:10490630

  8. The Rho-GEF Rom2p Localizes to Sites of Polarized Cell Growth and Participates in Cytoskeletal Functions in Saccharomyces cerevisiae

    PubMed Central

    Manning, Brendan D.; Padmanabha, Ramesh; Snyder, Michael

    1997-01-01

    Rom2p is a GDP/GTP exchange factor for Rho1p and Rho2p GTPases; Rho proteins have been implicated in control of actin cytoskeletal rearrangements. ROM2 and RHO2 were identified in a screen for high-copy number suppressors of cik1Δ, a mutant defective in microtubule-based processes in Saccharomyces cerevisiae. A Rom2p::3XHA fusion protein localizes to sites of polarized cell growth, including incipient bud sites, tips of small buds, and tips of mating projections. Disruption of ROM2 results in temperature-sensitive growth defects at 11°C and 37°C. rom2Δ cells exhibit morphological defects. At permissive temperatures, rom2Δ cells often form elongated buds and fail to form normal mating projections after exposure to pheromone; at the restrictive temperature, small budded cells accumulate. High-copy number plasmids containing either ROM2 or RHO2 suppress the temperature-sensitive growth defects of cik1Δ and kar3Δ strains. KAR3 encodes a kinesin-related protein that interacts with Cik1p. Furthermore, rom2Δ strains exhibit increased sensitivity to the microtubule depolymerizing drug benomyl. These results suggest a role for Rom2p in both polarized morphogenesis and functions of the microtubule cytoskeleton. PMID:9348527

  9. A microfluidic-based genetic screen to identify microbial virulence factors that inhibit dendritic cell migration

    PubMed Central

    McLaughlin, Laura M.; Xu, Hui; Carden, Sarah E.; Fisher, Samantha; Reyes, Monique; Heilshorn, Sarah C.; Monack, Denise M.

    2014-01-01

    Microbial pathogens are able to modulate host cells and evade the immune system by multiple mechanisms. For example, Salmonella injects effector proteins into host cells and evades the host immune system in part by inhibiting dendritic cell (DC) migration. The identification of microbial factors that modulate normal host functions should lead to the development of new classes of therapeutics that target these pathways. Current screening methods to identify either host or pathogen genes involved in modulating migration towards a chemical signal are limited because they do not employ stable, precisely controlled chemical gradients. Here, we develop a positive selection microfluidic-based genetic screen that allows us to identify Salmonella virulence factors that manipulate DC migration within stable, linear chemokine gradients. Our screen identified 7 Salmonella effectors (SseF, SifA, SspH2, SlrP, PipB2, SpiC and SseI) that inhibit DC chemotaxis toward CCL19. This method is widely applicable for identifying novel microbial factors that influence normal host cell chemotaxis as well as revealing new mammalian genes involved in directed cell migration. PMID:24599496

  10. A microfluidic-based genetic screen to identify microbial virulence factors that inhibit dendritic cell migration.

    PubMed

    McLaughlin, Laura M; Xu, Hui; Carden, Sarah E; Fisher, Samantha; Reyes, Monique; Heilshorn, Sarah C; Monack, Denise M

    2014-04-01

    Microbial pathogens are able to modulate host cells and evade the immune system by multiple mechanisms. For example, Salmonella injects effector proteins into host cells and evades the host immune system in part by inhibiting dendritic cell (DC) migration. The identification of microbial factors that modulate normal host functions should lead to the development of new classes of therapeutics that target these pathways. Current screening methods to identify either host or pathogen genes involved in modulating migration towards a chemical signal are limited because they do not employ stable, precisely controlled chemical gradients. Here, we develop a positive selection microfluidic-based genetic screen that allows us to identify Salmonella virulence factors that manipulate DC migration within stable, linear chemokine gradients. Our screen identified 7 Salmonella effectors (SseF, SifA, SspH2, SlrP, PipB2, SpiC and SseI) that inhibit DC chemotaxis toward CCL19. This method is widely applicable for identifying novel microbial factors that influence normal host cell chemotaxis as well as revealing new mammalian genes involved in directed cell migration. PMID:24599496

  11. Microbial Fuel Cells Applied to the Metabolically Based Detection of Extraterrestrial Life

    NASA Astrophysics Data System (ADS)

    Abrevaya, Ximena C.; Mauas, Pablo J. D.; Cortón, Eduardo

    2010-12-01

    Since the 1970s, when the Viking spacecrafts carried out experiments to detect microbial metabolism on the surface of Mars, the search for nonspecific methods to detect life in situ has been one of the goals of astrobiology. It is usually required that a methodology detect life independently from its composition or form and that the chosen biological signature point to a feature common to all living systems, such as the presence of metabolism. In this paper, we evaluate the use of microbial fuel cells (MFCs) for the detection of microbial life in situ. MFCs are electrochemical devices originally developed as power electrical sources and can be described as fuel cells in which the anode is submerged in a medium that contains microorganisms. These microorganisms, as part of their metabolic process, oxidize organic material, releasing electrons that contribute to the electric current, which is therefore proportional to metabolic and other redox processes. We show that power and current density values measured in MFCs that use microorganism cultures or soil samples in the anode are much larger than those obtained with a medium free of microorganisms or sterilized soil samples, respectively. In particular, we found that this is true for extremophiles, which have been proposed as potential inhabitants of extraterrestrial environments. Therefore, our results show that MFCs have the potential to be used for in situ detection of microbial life.

  12. Microbial fuel cells applied to the metabolically based detection of extraterrestrial life.

    PubMed

    Abrevaya, Ximena C; Mauas, Pablo J D; Cortón, Eduardo

    2010-12-01

    Since the 1970s, when the Viking spacecrafts carried out experiments to detect microbial metabolism on the surface of Mars, the search for nonspecific methods to detect life in situ has been one of the goals of astrobiology. It is usually required that a methodology detect life independently from its composition or form and that the chosen biological signature point to a feature common to all living systems, such as the presence of metabolism. In this paper, we evaluate the use of microbial fuel cells (MFCs) for the detection of microbial life in situ. MFCs are electrochemical devices originally developed as power electrical sources and can be described as fuel cells in which the anode is submerged in a medium that contains microorganisms. These microorganisms, as part of their metabolic process, oxidize organic material, releasing electrons that contribute to the electric current, which is therefore proportional to metabolic and other redox processes. We show that power and current density values measured in MFCs that use microorganism cultures or soil samples in the anode are much larger than those obtained with a medium free of microorganisms or sterilized soil samples, respectively. In particular, we found that this is true for extremophiles, which have been proposed as potential inhabitants of extraterrestrial environments. Therefore, our results show that MFCs have the potential to be used for in situ detection of microbial life. PMID:21162676

  13. Optofluidic Cell Selection from Complex Microbial Communities for Single-Genome Analysis

    PubMed Central

    Landry, Zachary C.; Giovanonni, Stephen J.; Quake, Stephen R.; Blainey, Paul C.

    2013-01-01

    Genetic analysis of single cells is emerging as a powerful approach for studies of heterogeneous cell populations. Indeed, the notion of homogeneous cell populations is receding as approaches to resolve genetic and phenotypic variation between single cells are applied throughout the life sciences. A key step in single-cell genomic analysis today is the physical isolation of individual cells from heterogeneous populations, particularly microbial populations, which often exhibit high diversity. Here, we detail the construction and use of instrumentation for optical trapping inside microfluidic devices to select individual cells for analysis by methods including nucleic acid sequencing. This approach has unique advantages for analyses of rare community members, cells with irregular morphologies, small quantity samples, and studies that employ advanced optical microscopy. PMID:24060116

  14. Quantifying the contribution of single microbial cells to nitrogen assimilation in aquatic environments

    NASA Astrophysics Data System (ADS)

    Musat, N.; Kuypers, M. M. M.

    2009-04-01

    Nitrogen is a primary productivity-limiting nutrient in the ocean. The nitrogen limitation of productivity may be overcome by organisms capable of converting dissolved N2 into fixed nitrogen available to the ecosystem. In many oceanic regions, growth of phytoplankton is nitrogen limited because fixation of N2 cannot make up for the removal of fixed inorganic nitrogen (NH4+, NO2-, NO3-) by anaerobic microbial processes. The amount of available fixed nitrogen in the ocean can be changed by the biological processes of heterotrophic denitrification, anaerobic ammonium oxidation and nitrogen fixation. For a complete understanding of nitrogen cycling in the ocean a link between the microbial and biogeochemical processes at the single cell level and their role in global biogeochemical cycles is essential. Here we report a recently developed method, Halogen In Situ Hybridization-Secondary Ion Mass Spectroscopy (HISH-SIMS) and its potential application to study the nitrogen-cycle processes in the ocean. The method allows simultaneous phylogenetic identification and quantitation of metabolic activities of single microbial cells in the environment. It uses horseradish-peroxidase-labeled oligonucleotide probes and fluorine-containing tyramides for the identification of microorganisms in combination with stable-isotope-labeling experiments for analyzing the metabolic function of single microbial cells. HISH-SIMS was successfully used to study nitrogen assimilation and nitrogen fixation by anaerobic phototrophs in a meromictic alpine lake. The HISH-SIMS method enables studies of the ecophysiology of individual, phylogenetically identified microorganisms involved in the N-cycle and allows us to track the flow of nitrogen within microbial communities.

  15. Bacterial Cellulose as a Substrate for Microbial Cell Culture

    PubMed Central

    Yin, Na; Santos, Thiago M. A.; Auer, George K.; Crooks, John A.; Oliver, Piercen M.

    2014-01-01

    Bacterial cellulose (BC) has a range of structural and physicochemical properties that make it a particularly useful material for the culture of bacteria. We studied the growth of 14 genera of bacteria on BC substrates produced by Acetobacter xylinum and compared the results to growth on the commercially available biopolymers agar, gellan, and xanthan. We demonstrate that BC produces rates of bacterial cell growth that typically exceed those on the commercial biopolymers and yields cultures with higher titers of cells at stationary phase. The morphology of the cells did not change during growth on BC. The rates of nutrient diffusion in BC being higher than those in other biopolymers is likely a primary factor that leads to higher growth rates. Collectively, our results suggest that the use of BC may open new avenues in microbiology by facilitating bacterial cell culture and isolation. PMID:24441155

  16. Overactivation of the protein kinase C-signaling pathway suppresses the defects of cells lacking the Rho3/Rho4-GAP Rgd1p in Saccharomyces cerevisiae.

    PubMed Central

    de Bettignies, G; Thoraval, D; Morel, C; Peypouquet, M F; Crouzet, M

    2001-01-01

    The nonessential RGD1 gene encodes a Rho-GTPase activating protein for the Rho3 and Rho4 proteins in Saccharomyces cerevisiae. Previous studies have revealed genetic interactions between RGD1 and the SLG1 and MID2 genes, encoding two putative sensors for cell integrity signaling, and VRP1 encoding an actin and myosin interacting protein involved in polarized growth. To better understand the role of Rgd1p, we isolated multicopy suppressor genes of the cell lethality of the double mutant rgd1Delta mid2Delta. RHO1 and RHO2 encoding two small GTPases, MKK1 encoding one of the MAP-kinase kinases in the protein kinase C (PKC) pathway, and MTL1, a MID2-homolog, were shown to suppress the rgd1Delta defects strengthening the functional links between RGD1 and the cell integrity pathway. Study of the transcriptional activity of Rlm1p, which is under the control of Mpk1p, the last kinase of the PKC pathway, and follow-up of the PST1 transcription, which is positively regulated by Rlm1p, indicate that the lack of RGD1 function diminishes the PKC pathway activity. We hypothesize that the rgd1Delta inactivation, at least through the hyperactivation of the small GTPases Rho3p and Rho4p, alters the secretory pathway and/or the actin cytoskeleton and decreases activity of the PKC pathway. PMID:11779787

  17. Promotion of Cell Viability and Histone Gene Expression by the Acetyltransferase Gcn5 and the Protein Phosphatase PP2A in Saccharomyces cerevisiae.

    PubMed

    Petty, Emily L; Lafon, Anne; Tomlinson, Shannon L; Mendelsohn, Bryce A; Pillus, Lorraine

    2016-08-01

    Histone modifications direct chromatin-templated events in the genome and regulate access to DNA sequence information. There are multiple types of modifications, and a common feature is their dynamic nature. An essential step for understanding their regulation, therefore, lies in characterizing the enzymes responsible for adding and removing histone modifications. Starting with a dosage-suppressor screen in Saccharomyces cerevisiae, we have discovered a functional interaction between the acetyltransferase Gcn5 and the protein phosphatase 2A (PP2A) complex, two factors that regulate post-translational modifications. We find that RTS1, one of two genes encoding PP2A regulatory subunits, is a robust and specific high-copy suppressor of temperature sensitivity of gcn5∆ and a subset of other gcn5∆ phenotypes. Conversely, loss of both PP2A(Rts1) and Gcn5 function in the SAGA and SLIK/SALSA complexes is lethal. RTS1 does not restore global transcriptional defects in gcn5∆; however, histone gene expression is restored, suggesting that the mechanism of RTS1 rescue includes restoration of specific cell cycle transcripts. Pointing to new mechanisms of acetylation-phosphorylation cross-talk, RTS1 high-copy rescue of gcn5∆ growth requires two residues of H2B that are phosphorylated in human cells. These data highlight the potential significance of dynamic phosphorylation and dephosphorylation of these deeply conserved histone residues for cell viability. PMID:27317677

  18. Increase in chitin as an essential response to defects in assembly of cell wall polymers in the ggp1delta mutant of Saccharomyces cerevisiae.

    PubMed Central

    Popolo, L; Gilardelli, D; Bonfante, P; Vai, M

    1997-01-01

    The GGP1/GAS1 gene codes for a glycosylphosphatidylinositol-anchored plasma membrane glycoprotein of Saccharomyces cerevisiae. The ggp1delta mutant shows morphogenetic defects which suggest changes in the cell wall matrix. In this work, we have investigated cell wall glucan levels and the increase of chitin in ggp1delta mutant cells. In these cells, the level of alkali-insoluble 1,6-beta-D-glucan was found to be 50% of that of wild-type cells and was responsible for the observed decrease in the total alkali-insoluble glucan. Moreover, the ratio of alkali-soluble to alkali-insoluble glucan almost doubled, suggesting a change in glucan solubility. The increase of chitin in ggp1delta cells was found to be essential since the chs3delta ggp1delta mutations determined a severe reduction in the growth rate and in cell viability. Electron microscopy analysis showed the loss of the typical structure of yeast cell walls. Furthermore, in the chs3delta ggp1delta cells, the level of alkali-insoluble glucan was 57% of that of wild-type cells and the alkali-soluble/alkali-insoluble glucan ratio was doubled. We tested the effect of inhibition of chitin synthesis also by a different approach. The ggp1delta cells were treated with nikkomycin Z, a well-known inhibitor of chitin synthesis, and showed a hypersensitivity to this drug. In addition, studies of genetic interactions with genes related to the construction of the cell wall indicate a synthetic lethal effect of the ggp1delta kre6delta and the ggp1delta pkc1delta combined mutations. Our data point to an involvement of the GGP1 gene product in the cross-links between cell wall glucans (1,3-beta-D-glucans with 1,6-beta-D-glucans and with chitin). Chitin is essential to compensate for the defects due to the lack of Ggp1p. Moreover, the activities of Ggp1p and Chs3p are essential to the formation of the organized structure of the cell wall in vegetative cells. PMID:8990299

  19. Distinct roles of T-cell lymphopenia and the microbial flora for gastrointestinal and CNS autoimmunity.

    PubMed

    Fischer, Henrike J; Witte, Ann-Kathrin; Walter, Lutz; Gröne, Hermann-Josef; van den Brandt, Jens; Reichardt, Holger M

    2016-05-01

    T-cell lymphopenia is a major risk factor for autoimmunity. Here we describe congenic Lewis (LEW) rats with a loss-of-function mutation in the Gimap5 gene, leading to a 92% reduction in peripheral T-cell numbers. Gimap5-deficient LEW rats developed eosinophilic autoimmune gastroenteritis accompanied by a 40-fold increase in IgE serum levels. This phenotype was ameliorated by antibiotic treatment, indicating a critical role of the microbial flora in the development of inflammatory bowel disease. Interestingly, Gimap5-deficient LEW rats showed strongly aggravated experimental autoimmune encephalomyelitis (EAE) after immunization with guinea pig myelin basic protein. This phenotype, however, persisted after antibiosis, confirming that the enhanced CNS autoimmune response in T-cell lymphopenic Gimap5-deficient LEW rats was unrelated to the composition of the microbial flora. Rather, it seems that it was caused by the 7-fold increase in the percentage of activated T cells producing IL-17 and IFN-γ, and the skewed T-cell receptor (TCR) repertoire, both of which were the result of T-cell lymphopenia and not affected by antibiosis. This notion was supported by the observation that adoptive T-cell transfer corrected the TCR repertoire and improved EAE. Collectively, our findings confirm a critical albeit differential role of T-cell lymphopenia in the susceptibility to organ-specific autoimmune responses.-Fischer, H. J., Witte, A.-K., Walter, L., Gröne, H.-J., van den Brandt, J., Reichardt, H. M. Distinct roles of T-cell lymphopenia and the microbial flora for gastrointestinal and CNS autoimmunity. PMID:26740263

  20. Electricity and H2 generation from hemicellulose by sequential fermentation and microbial fuel/electrolysis cell

    NASA Astrophysics Data System (ADS)

    Yan, Di; Yang, Xuewei; Yuan, Wenqiao

    2015-09-01

    Electricity and hydrogen generation by bacteria Geobacter sulfurreducens in a dual-chamber microbial fuel/electrolysis cell following the fermentation of hemicellulose by bacteria Moorella thermoacetica was investigated. Experimental results showed that 10 g l-1 xylose under 60 °C was appropriate for the fermentation of xylose by M. thermoacetica, yielding 0.87 g-acetic acid per gram of xylose consumed. Corncob hydrolysate could also be fermented to produce acetic acid, but with lower yield (0.74 g-acid per g-xylose). The broths of xylose and corncob hydrolysate fermented by M. thermoacetica containing acetic acid were fed to G. sulfurreducens in a dual-chamber microbial fuel/electrolysis cell for electricity and hydrogen generation. The highest open-circuit cell voltages generated were 802 and 745 mV, and hydrogen yields were 41.7 and 23.3 mmol per mol-acetate, in xylose and corncob hydrolysate fermentation broth media, respectively. The internal resistance of the microbial fuel/electrolysis cell fed with corncob hydrolysate fermentation broth (3472 Ω) was much higher than that with xylose fermentation broth (1993 Ω) or sodium acetate medium (467 Ω), which was believed to be the main cause of the variation in hydrogen yield of the three feeding media.

  1. Surface display of malolactic enzyme from Oenococcus oeni on Saccharomyces cerevisiae.

    PubMed

    Zhang, Xiuyan; Hou, Xiaoyan; Liang, Fang; Chen, Fusheng; Wang, Xiaohong

    2013-04-01

    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. PMID:23446978

  2. Association of Transcription Factor IIA with TATA Binding Protein Is Required for Transcriptional Activation of a Subset of Promoters and Cell Cycle Progression in Saccharomyces cerevisiae

    PubMed Central

    Ozer, Josef; Lezina, Larissa E.; Ewing, Joshua; Audi, Salma; Lieberman, Paul M.

    1998-01-01

    The general transcription factor IIA (TFIIA) interacts with the TATA binding protein (TBP) and promoter DNA to mediate transcription activation in vitro. To determine if this interaction is generally required for activation of all class II genes in vivo, we have constructed substitution mutations in yeast TFIIA which compromise its ability to bind TBP. Substitution mutations in the small subunit of TFIIA (Toa2) at residue Y69 or W76 significantly impaired the ability of TFIIA to stimulate TBP-promoter binding in vitro. Gene replacement of wild-type TOA2 with a W76E or Y69A/W76A mutant was lethal in Saccharomyces cerevisiae, while the Y69F/W76F mutant exhibited extremely slow growth at 30°C. Both the Y69A and W76A mutants were conditionally lethal at higher temperatures. Light microscopy indicated that viable toa2 mutant strains accumulate as equal-size dumbbells and multibudded clumps. Transcription of the cell cycle-regulatory genes CLB1, CLB2, CLN1, and CTS1 was significantly reduced in the toa2 mutant strains, while the noncycling genes PMA1 and ENO2 were only modestly affected, suggesting that these toa2 mutant alleles disrupt cell cycle progression. The differential effect of these toa2 mutants on gene transcription was examined for a number of other genes. toa2 mutant strains supported high levels of CUP1, PHO5, TRP3, and GAL1 gene activation, but the constitutive expression of DED1 was significantly reduced. Activator-induced start site expression for HIS3, GAL80, URA1, and URA3 promoters was defective in toa2 mutant strains, suggesting that the TFIIA-TBP complex is important for promoters which require an activator-dependent start site selection from constitutive to regulated expression. We present evidence to indicate that transcription defects in toa2 mutants can be both activator and promoter dependent. These results suggest that the association of TFIIA with TBP regulates activator-induced start site selection and cell cycle progression in S

  3. [Tolerance of Saccharomyces cerevisiae to monoterpenes--a review].

    PubMed

    Liu, Jidong; Zhou, Jingwen; Chen, Jian

    2013-06-01

    Tolerance of Saccharomyces cerevisiae to monoterpenes is important in both metabolic engineering of the yeast to produce these chemicals de novo and efficient use of biomass containing these chemicals. Understanding the mechanisms in the tolerance of S. cerevisiae to monoterpenes could facilitate the construction of yeast strains with enhanced monoterpenes resistance, and therefore improve related bioprocesses. Monoterpenes could disturb the redox balance in S. cerevisiae, therefore increase the accumulation of reactive oxygen species (ROS) and result in cell death. S. cerevisiae has to systematically improve its antioxidative ability to deal with the ROS induced damage. The current review summarized the recent developments in demonstration of the tolerance of S. cerevisiae to different typical monoterpenes mainly from the aspect of the antioxidative mechanisms. Based on the analysis of the previous works, further attempts to demonstrate the mechanisms were proposed. PMID:24028054

  4. [Determination of Azospirillum Brasilense Cells With Bacteriophages via Electrooptical Analysis of Microbial Suspensions].

    PubMed

    Gulii, O I; Karavayeva, O A; Pavlii, S A; Sokolov, O I; Bunin, V D; Ignatov, O V

    2015-01-01

    The dependence-of changes in the electrooptical properties of Azospirillum brasilense cell suspension Sp7 during interaction with bacteriophage ΦAb-Sp7 on the number and time of interactions was studied. Incubation of cells with bacteriophage significantly changed the electrooptical signal within one minute. The selective effect of bacteriophage ΦAb on 18 strains of bacteria of the genus Azospirillum was studied: A. amazonense Ami4, A. brasilense Sp7, Cd, Sp107, Sp245, Jm6B2, Brl4, KR77, S17, S27, SR55, SR75, A. halopraeferans Au4, A. irakense KBC1, K A3, A. lipoferum Sp59b, SR65 and RG20a. We determined the limit of reliable determination of microbial cells infected with bacteriophage: - 10(4) cells/mL. The presence of foreign cell cultures of E. coli B-878 and E. coli XL-1 did not complicate the detection of A brasilense Sp7 cells with the use of bacteriophage ΦAb-Sp7. The results demonstrated that bacteriophage (ΦAb-Sp7 can be used for the detection of Azospirillum microbial cells via t electrooptical analysis of cell suspensions. PMID:26204775

  5. Alternate switching between microbial fuel cell and microbial electrolysis cell operation as a new method to control H2O2 level in Bioelectro-Fenton system

    NASA Astrophysics Data System (ADS)

    Zhang, Yifeng; Wang, Yong; Angelidaki, Irini

    2015-09-01

    Sustainable H2O2 supply and cost-effective elimination of residual H2O2 are two key challenges associated with the successful application of Fenton reaction for contaminant removal. In this study, an innovative Bioelectro-Fenton system capable of alternate switching between microbial electrolysis cell (MEC) and microbial fuel cell (MFC) mode of operation was developed to meet the challenges. In the MEC mode, a bioelectrochemical system (BES) produces H2O2 which reacts with Fenton's reagent (Fe II) to form hydroxyradical. The unused H2O2 (residual H2O2) is removed as electron acceptor by switching the system to MFC mode of operation. Complete decolorization and mineralization of 50 mg L-1 methylene blue (MB) was achieved in the MEC mode with apparent first order rate constants of 0.43 and 0.22 h-1, respectively. After switching to the MFC mode, residual H2O2 of 180 mg L-1 was removed at a removal rate of 4.61 mg L-1 h-1 while generating a maximum current density of 0.49 A m-2. The MB degradation and residual H2O2 removal were affected by external resistance, cathode pH and initial MB concentration. Furthermore, the system performance was enhanced under stack operation. This study provides a proof-in-concept new system for efficient and cost-effective H2O2 control and recalcitrant pollutants removal.

  6. Two-stage conversion of crude glycerol to energy using dark fermentation linked with microbial fuel cell or microbial electrolysis cell.

    PubMed

    Chookaew, Teera; Prasertsan, Poonsuk; Ren, Zhiyong Jason

    2014-03-25

    Crude glycerol is a main byproduct of the biodiesel industry, and the beneficial use of waste glycerol has been a major challenge. This study characterises the conversion of crude glycerol into bioenergy such as H2 and electricity using a two-stage process linking dark fermentation with a microbial fuel cell (MFC) or microbial electrolysis cell (MEC). The results showed that fermentation achieved a maximum H2 rate of 332 mL/L and a yield of 0.55 mol H2/mol glycerol, accompanied by 20% of organic removal. Fed with the raw fermentation products with an initial COD of 7610 mg/L, a two-chamber MFC produced 92 mW/m(2) in power density and removed 50% of COD. The Columbic efficiency was 14%. When fed with 50% diluted fermentation product, a similar power output (90m W/m(2)) and COD removal (49%) were obtained, but the CE doubled to 27%. Similar substrates were used to produce H2 in two-chamber MECs, and the diluted influent had a higher performance, with the highest yield at 106 mL H2/g COD and a CE of 24%. These results demonstrate that dark fermentation linked with MFC/MEC can be a feasible option for conversion of waste glycerol into bioenergy. PMID:24380781

  7. The Cdc42 GTPase-associated proteins Gic1 and Gic2 are required for polarized cell growth in Saccharomyces cerevisiae

    PubMed Central

    Chen, Guang-Chao; Kim, Yung-Jin; Chan, Clarence S.M.

    1997-01-01

    BEM2 of Saccharomyces cerevisiae encodes a Rho-type GTPase-activating protein that is required for proper bud site selection at 26°C and for bud emergence at elevated temperatures. We show here that the temperature-sensitive growth phenotype of bem2 mutant cells can be suppressed by increased dosage of the GIC1 gene. The Gic1 protein, together with its structural homolog Gic2, are required for cell size and shape control, bud site selection, bud emergence, actin cytoskeletal organization, mitotic spindle orientation/positioning, and mating projection formation in response to mating pheromone. Each protein contains a CRIB (Cdc42/Rac-interactive binding) motif and each interacts in the two-hybrid assay with the GTP-bound form of the Rho-type Cdc42 GTPase, a key regulator of polarized growth in yeast. The CRIB motif of Gic1 and the effector domain of Cdc42 are required for this association. Genetic experiments indicate that Gic1 and Gic2 play positive roles in the Cdc42 signal transduction pathway, probably as effectors of Cdc42. Subcellular localization studies with a functional green fluorescent protein–Gic1 fusion protein indicate that this protein is concentrated at the incipient bud site of unbudded cells, at the bud tip and mother-bud neck of budded cells, and at cortical sites on large-budded cells that may delimit future bud sites in the two progeny cells. The ability of Gic1 to associate with Cdc42 is important for its function but is apparently not essential for its subcellular localization. PMID:9367979

  8. (R)-PAC biosynthesis in [BMIM][PF₆]/aqueous biphasic system using Saccharomyces cerevisiae BY4741 cells.

    PubMed

    Kandar, Smita; Suresh, A K; Noronha, Santosh B

    2015-02-01

    (R)-phenylacetylcarbinol or (R)-PAC is a pharmaceutical precursor of (1R, 2S) ephedrine and (1S, 2S) pseudoephedrine. Biotransformation of benzaldehyde and glucose by pyruvate decarboxylase produces (R)-PAC. This biotransformation suffers from toxicity of the substrate, product [(R)-PAC] and by-product (benzyl alcohol). In the present study, ionic liquid/aqueous biphasic system was employed to enhance (R)-PAC production. Fermented broth was the reaction medium in which Saccharomyces cerevisiae BY4741 was the source of pyruvate decarboxylase. Hydrophobic ionic liquid 1-butyl-3-methylimidazolium hexafluorophosphate ([BMIM][PF6]) was the non-aqueous phase in which toxic compounds reside. Biocompatibility of [BMIM][PF6] and adequate distribution coefficients of benzaldehyde, (R)-PAC and benzyl alcohol were determined. A Box-Behnken design and response surface methodology were used for the optimization of biotransformation variables in order to maximize (R)-PAC yield and productivity. The results showed higher (R)-PAC yield and productivity of ∼1.5-fold each in the biphasic biotransformation of phase volume ratio 0.05 as compared to the monophasic (conventional) biotransformation. Moreover, the level of major by-product benzyl alcohol was also 3.5-fold lower in biphasic biotransformation. [BMIM][PF6]/aqueous biphasic system is a new approach which could intensify the (R)-PAC production. PMID:25424285

  9. Anaerobic and sequential aerobic production of high-titer ethanol and single cell protein from NaOH-pretreated corn stover by a genome shuffling-modified Saccharomyces cerevisiae strain.

    PubMed

    Ren, Xueliang; Wang, Juncong; Yu, Hui; Peng, Chunlan; Hu, Jinlong; Ruan, Zhiyong; Zhao, Shumiao; Liang, Yunxiang; Peng, Nan

    2016-10-01

    In this study, a Saccharomyces cerevisiae recombinant strain 14 was constructed through genome shuffling method by transferring the whole genomic DNA of Candida intermedia strain 23 into a thermo-tolerant S. cerevisiae strain. The recombinant strain 14 combined the good natures of both parent strains that efficiently produced ethanol from glucose and single cell protein from xylose with 54.6% crude protein and all essential amino acids except cysteine at 35°C. Importantly, the recombinant strain 14 produced 64.07g/L ethanol from 25%(w/v) NaOH-pretreated and washed corn stover with the ethanol yield of 0.26g/g total stover by fed-batch simultaneous saccharification and fermentation and produced 66.50g/L dry cell mass subsequently from the residual hydrolysate and ethanol. Therefore, this study represents a feasible method to comprehensively utilize hexose and pentose in lignocellulosic materials. PMID:27416512

  10. Electricity generation from cattle dung using microbial fuel cell technology during anaerobic acidogenesis and the development of microbial populations.

    PubMed

    Zhao, Guang; Ma, Fang; Wei, Li; Chua, Hong; Chang, Chein-Chi; Zhang, Xiao-Jun

    2012-09-01

    A microbial fuel cell (MFC) was constructed to investigate the possible generation of electricity using cattle dung as a substrate. After 30 days of operation, stable electricity was generated, and the maximum volumetric power density was 0.220 W/m(3). The total chemical oxygen demand (TCOD) removal and coulombic efficiency (CE) of the MFC reached 73.9±1.8% and 2.79±0.6%, respectively, after 120 days of operation. Acetate was the main metabolite in the anolyte, and other volatile fatty acids (VFAs) (propionate and butyrate) were present in minor amounts. The PCR-DGGE analysis indicated that the following five groups of microbes were present: Proteobacteria, Bacteroides, Chloroflexi, Actinobacteria and Firmicutes. Proteobacteria and Firmicutes were the dominant phyla in the sample; specifically, 36.3% and 24.2% of the sequences obtained were Proteobacteria and Firmicutes, respectively. Clostridium sp., Pseudomonas luteola and Ochrobactrum pseudogrignonense were the most dominant groups during the electricity generation process. The diversity of archaea dramatically decreased after 20 days of operation. The detected archaea were hydrogenotrophic methanogens, and the Methanobacterium genus disappeared during the periods of stable electricity generation via acidogenesis. PMID:22595839

  11. Characterization of methane production and microbial community shifts during waste activated sludge degradation in microbial electrolysis cells.

    PubMed

    Sun, Rui; Zhou, Aijuan; Jia, Jianna; Liang, Qing; Liu, Qian; Xing, Defeng; Ren, Nanqi

    2015-01-01

    Microbial electrolysis cell (MECs) were investigated as a promising technology to manage waste activated sludge (WAS) reduction and bio-methane generation. The effect of WAS concentration on the MECs performance was discussed. At the optimal concentration of 15gCOD/L, maximum methane yield of MECs fed with alkaline pretreated WAS (A-WAS) were achieved with the value of 77.13±2.52LCH4/kg-COD on Day 3, which had been improved by 1.5-fold compared with MECs fed with raw WAS (R-WAS), while that was negligible in open circuit controls. Efficient sludge reduction was also obtained in terms of TCOD, total protein, TSS and VSS removal. Pyrosequencing revealed the dominance of exoelectrogen Geobacter and hydrogen-producing bacteria Petrimonas in MECs fed with WAS. Methanocorpusculum with the capacity of methane generation using CO2 and H2 also showed overwhelming dominance (96.01%). The large proportions of Petrimonas and Methanocorpusculum indicated the occurrence of hydrogenotrophic methanogenesis in our methane-producing MECs. PMID:25459805

  12. Bifunctional silver nanoparticle cathode in microbial fuel cells for microbial growth inhibition with comparable oxygen reduction reaction activity.

    PubMed

    An, Junyeong; Jeon, Hongrae; Lee, Jaeyoung; Chang, In Seop

    2011-06-15

    Organic contamination of water bodies in which benthic microbial fuel cells (benthic MFCs) are installed, and organic crossover from the anode to the cathode of membraneless MFCs, is a factor causing oxygen depletion and substrate loss in the cathode due to the growth of heterotrophic aerobic bacteria. This study examines the possible use of silver nanoparticles (AgNPs) as a cathodic catalyst for MFCs suffering from organic contamination and oxygen depletion. Four treated cathodes (AgNPs-coated, Pt/C-coated, Pt/C+AgNPs-coated, and plain graphite cathodes) were prepared and tested under high levels of organics loading. During operation (fed with 50 mM acetate), the AgNPs-coated system showed the highest DO concentration (0.8 mg/L) in the cathode area as well as the highest current (ranging from 0.04 to 0.12 mA). Based on these results, we concluded that (1) the growth of oxygen-consuming heterotrophic microbes could be inhibited by AgNPs, (2) the function of AgNPs as a bacterial growth inhibitor resulted in a greater increase of DO concentration in the cathode than the other tested cathode systems, (3) AgNPs could be applied as a cathode catalyst for oxygen reduction, and as a result (4) the MFC with the AgNPs-coated cathode led to the highest current generation among the tested MFCs. PMID:21585217

  13. Operational, design and microbial aspects related to power production with microbial fuel cells implemented in constructed wetlands.

    PubMed

    Corbella, Clara; Guivernau, Miriam; Viñas, Marc; Puigagut, Jaume

    2015-11-01

    This work aimed at determining the amount of energy that can be harvested by implementing microbial fuel cells (MFC) in horizontal subsurface constructed wetlands (HSSF CWs) during the treatment of real domestic wastewater. To this aim, MFC were implemented in a pilot plant based on two HSSF CW, one fed with primary settled wastewater (Settler line) and the other fed with the effluent of a hydrolytic up-flow sludge blanket reactor (HUSB line). The eubacterial and archaeal community was profiled on wetland gravel, MFC electrodes and primary treated wastewater by means of 16S rRNA gene-based 454-pyrosequencing and qPCR of 16S rRNA and mcrA genes. Maximum current (219 mA/m(2)) and power (36 mW/m(2)) densities were obtained for the HUSB line. Power production pattern correlated well with water level fluctuations within the wetlands, whereas the type of primary treatment implemented had a significant impact on the diversity and relative abundance of eubacteria communities colonizing MFC. It is worth noticing the high predominance (13-16% of relative abundance) of one OTU belonging to Geobacter on active MFC of the HUSB line that was absent for the settler line MFC. Hence, MFC show promise for power production in constructed wetlands receiving the effluent of a HUSB reactor. PMID:26253894

  14. Power densities and microbial communities of brewery wastewater-fed microbial fuel cells according to the initial substrates.

    PubMed

    Yu, Jaecheul; Park, Younghyun; Kim, Byunggoon; Lee, Taeho

    2015-01-01

    Single-chamber microbial fuel cells (MFCs) acclimated with glucose, butyrate, propionate, acetate, and a mixture of the four were operated with brewery wastewater (BWW) under a fed-batch mode. Glucose-fed MFC (GW-MFC) showed the highest maximum power density (PDmax) of 1,519 mW/m(2), followed in order by acetate-fed MFC (AW-MFC), mixed substrates-fed MFC (MW-MFC), butyrate-fed MFC (BW-MFC), and propionate-fed MFC (PW-MFC). After changing to BWW, power production was decreased for all MFCs. MFC acclimated with glucose showed the highest PDmax of 890 ± 12 mW/m(2), followed in order by MW-MFC, AW-MFC, BW-MFC, and PW-MFC. The PDmax in BWW-MFC, which was acclimated and operated with BWW, of 552 mW/m(2) was less than that of GW-MFC and MW-MFC but more than that of AW-MFC, BW-MFC, and PW-MFC. MFCs with fermentable substrates were less affected by the BWW. Gammaproteobacteria, including Pseudomonas sp., Acinetobacter sp. and Xanthomonas axonopodis, were found in all MFCs with pure substrates and Rubrivivax benzoatilyticus, thiobacillus sp. and Denitratisoma oestradiolicum belonging to Betaproteobacteria were newly detected in all MFCs when the substrate was changed to BWW. PMID:24973903

  15. Recombinant pharmaceuticals from microbial cells: a 2015 update.

    PubMed

    Sanchez-Garcia, Laura; Martín, Lucas; Mangues, Ramon; Ferrer-Miralles, Neus; Vázquez, Esther; Villaverde, Antonio

    2016-01-01

    Diabetes, growth or clotting disorders are among the spectrum of human diseases related to protein absence or malfunction. Since these pathologies cannot be yet regularly treated by gene therapy, the administration of functional proteins produced ex vivo is required. As both protein extraction from natural producers and chemical synthesis undergo inherent constraints that limit regular large-scale production, recombinant DNA technologies have rapidly become a choice for therapeutic protein production. The spectrum of organisms exploited as recombinant cell factories has expanded from the early predominating Escherichia coli to alternative bacteria, yeasts, insect cells and especially mammalian cells, which benefit from metabolic and protein processing pathways similar to those in human cells. Up to date, around 650 protein drugs have been worldwide approved, among which about 400 are obtained by recombinant technologies. Other 1300 recombinant pharmaceuticals are under development, with a clear tendency towards engineered versions with improved performance and new functionalities regarding the conventional, plain protein species. This trend is exemplified by the examination of the contemporary protein-based drugs developed for cancer treatment. PMID:26861699

  16. Increased performance of hydrogen production in microbial electrolysis cells under alkaline conditions.

    PubMed

    Rago, Laura; Baeza, Juan A; Guisasola, Albert

    2016-06-01

    This work reports the first successful enrichment and operation of alkaline bioelectrochemical systems (microbial fuel cells, MFC, and microbial electrolysis cells, MEC). Alkaline (pH=9.3) bioelectrochemical hydrogen production presented better performance (+117%) compared to conventional neutral conditions (2.6 vs 1.2 litres of hydrogen gas per litre of reactor per day, LH2·L(-1)REACTOR·d(-1)). Pyrosequencing results of the anodic biofilm showed that while Geobacter was mainly detected under conventional neutral conditions, Geoalkalibacter sp. was highly detected in the alkaline MFC (21%) and MEC (48%). This is the first report of a high enrichment of Geoalkalibacter from an anaerobic mixed culture using alkaline conditions in an MEC. Moreover, Alkalibacter sp. was highly present in the anodic biofilm of the alkaline MFC (37%), which would indicate its potentiality as a new exoelectrogen. PMID:26855359

  17. Biofilm vivacity and destruction on antimicrobial nanosurfaces assayed within a microbial fuel cell.

    PubMed

    Sugnaux, Marc; Fischer, Fabian

    2016-08-01

    A novel method was developed to assay the antimicrobial capacity of nanostructured surfaces for medical implants in a bicathodic microbial fuel cell. Nano-structured gold surfaces with protruding nanopillars and nanorings were investigated. Escherichia coli K12 were used as a model microbe to record electronic effects caused by the interaction with nanosurfaces. The nanostructured gold surfaces enabled power density maxima up to 1910mW/m(2), indicating fair vivacity, while flat surfaces on the nanoscale provided almost no power 0.35mW/m(2). The biofilm presence on antimicrobial nanosurfaces was confirmed by the addition of ampicillin and its bactericidal effect resulted in oscillating and declining potentiometric signals. Current density experiments showed that biofilms on antimicrobial nanostructured electrodes caused low currents, indicating that E.coli biofilm remained functional before destruction. The bicathodic microbial fuel cell sensor is a novel tool for evaluating antimicrobial effects caused by nanosurfaces and antibiotics. PMID:27071334

  18. Application of genetically engineered microbial whole-cell biosensors for combined chemosensing.

    PubMed

    He, Wei; Yuan, Sheng; Zhong, Wen-Hui; Siddikee, Md Ashaduzzaman; Dai, Chuan-Chao

    2016-02-01

    The progress of genetically engineered microbial whole-cell biosensors for chemosensing and monitoring has been developed in the last 20 years. Those biosensors respond to target chemicals and produce output signals, which offer a simple and alternative way of assessment approaches. As actual pollution caused by human activities usually contains a combination of different chemical substances, how to employ those biosensors to accurately detect real contaminant samples and evaluate biological effects of the combined chemicals has become a realistic object of environmental researches. In this review, we outlined different types of the recent method of genetically engineered microbial whole-cell biosensors for combined chemical evaluation, epitomized their detection performance, threshold, specificity, and application progress that have been achieved up to now. We also discussed the applicability and limitations of this biosensor technology and analyzed the optimum conditions for their environmental assessment in a combined way. PMID:26615397

  19. Quantification of effective exoelectrogens by most probable number (MPN) in a microbial fuel cell.

    PubMed

    Heidrich, Elizabeth S; Curtis, Thomas P; Woodcock, Stephen; Dolfing, Jan

    2016-10-01

    The objective of this work was to quantify the number of exoelectrogens in wastewater capable of producing current in a microbial fuel cell by adapting the classical most probable number (MPN) methodology using current production as end point. Inoculating a series of microbial fuel cells with various dilutions of domestic wastewater and with acetate as test substrate yielded an apparent number of exoelectrogens of 17perml. Using current as a proxy for activity the apparent exoelectrogen growth rate was 0.03h(-1). With starch or wastewater as more complex test substrates similar apparent growth rates were obtained, but the apparent MPN based numbers of exoelectrogens in wastewater were significantly lower, probably because in contrast to acetate, complex substrates require complex food chains to deliver the electrons to the electrodes. Consequently, the apparent MPN is a function of the combined probabilities of members of the food chain being present. PMID:27347794

  20. Power generation enhancement in novel microbial carbon capture cells with immobilized Chlorella vulgaris

    NASA Astrophysics Data System (ADS)

    Zhou, Minghua; He, Huanhuan; Jin, Tao; Wang, Hongyu

    2012-09-01

    With the increasing concerns for global climate change, a sustainable, efficient and renewable energy production from wastewater is imperative. In this study, a novel microbial carbon capture cell (MCC), is constructed for the first time by the introduction of immobilized microalgae (Chlorella vulgaris) into the cathode chamber of microbial fuel cells (MFCs) to fulfill the zero discharge of carbon dioxide. This process can achieve an 84.8% COD removal, and simultaneously the maximum power density can reach 2485.35 mW m-3 at a current density of 7.9 A m-3 and the Coulombic efficiency is 9.40%, which are 88% and 57.7% greater than that with suspended C. vulgaris, respectively. These enhancements in performance demonstrate the feasibility of an economical and effective approach for the simultaneous wastewater treatment, electricity generation and biodiesel production from microalgae.

  1. Novel microbial fuel cell design to operate with different wastewaters simultaneously.

    PubMed

    Mathuriya, Abhilasha Singh

    2016-04-01

    A novel single cathode chamber and multiple anode chamber microbial fuel cell design (MAC-MFC) was developed by incorporating multiple anode chambers into a single unit and its performance was checked. During 60 days of operation, performance of MAC-MFC was assessed and compared with standard single anode/cathode chamber microbial fuel cell (SC-MFC). The tests showed that MAC-MFC generated stable and higher power outputs compared with SC-MFC and each anode chamber contributed efficiently. Further, MAC-MFCs were incorporated with different wastewaters in different anode chambers and their behavior in MFC performance was observed. MAC-MFC efficiently treated multiple wastewaters simultaneously at low cost and small space, which claims its candidature for future possible scale-up applications. PMID:27090700

  2. Characteristic changes in algal organic matter derived from Microcystis aeruginosa in microbial fuel cells.

    PubMed

    Wang, Huan; Lu, Lu; Liu, Dongmei; Cui, Fuyi; Wang, Peng

    2015-11-01

    The objective of this study was to investigate behavior of algal organic matter (AOM) during bioelectrochemical oxidation in microbial fuel cell in terms of compositions and structures. Study revealed that the AOM derived from blue-green algae Microcystis aeruginosa could be degraded more completely (82% COD removal) in microbial fuel cells (MFCs) than by anaerobic fermentation (24% COD removal) in a control reactor without closed-circuit electrode and electricity was produced simultaneously. A variety of techniques were used to characterize the changes in AOM compositions and structures during bioelectrochemical oxidation. The presence of syntrophic interactions between electrochemical active bacteria and fermentative bacteria to degrade large molecular organics into small molecular substances, which could be oxidized by electrode but not by fermentation. The dominant tryptophan protein-like substances, humic acid-like substances and Chlorophyll a in AOM were highly degraded during MFC treatment. PMID:26081162

  3. An efficient approach to cathode operational parameters optimization for microbial fuel cell using response surface methodology

    PubMed Central

    2014-01-01

    Background In the recent study, optimum operational conditions of cathode compartment of microbial fuel cell were determined by using Response Surface Methodology (RSM) with a central composite design to maximize power density and COD removal. Methods The interactive effects of parameters such as, pH, buffer concentration and ionic strength on power density and COD removal were evaluated in two-chamber microbial batch-mode fuel cell. Results Power density and COD removal for optimal conditions (pH of 6.75, buffer concentration of 0.177 M and ionic strength of cathode chamber of 4.69 mM) improve by 17 and 5%, respectively, in comparison with normal conditions (pH of 7, buffer concentration of 0.1 M and ionic strength of 2.5 mM). Conclusions In conclusion, results verify that response surface methodology could successfully determine cathode chamber optimum operational conditions. PMID:24423039

  4. Oral microbial biofilm stimulation of epithelial cell responses.

    PubMed

    Peyyala, Rebecca; Kirakodu, Sreenatha S; Novak, Karen F; Ebersole, Jeffrey L

    2012-04-01

    Oral bacterial biofilms trigger chronic inflammatory responses in the host that can result in the tissue destructive events of periodontitis. However, the characteristics of the capacity of specific host cell types to respond to these biofilms remain ill-defined. This report describes the use of a novel model of bacterial biofilms to stimulate oral epithelial cells and profile select cytokines and chemokines that contribute to the local inflammatory environment in the periodontium. Monoinfection biofilms were developed with Streptococcus sanguinis, Streptococcus oralis, Streptococcus gordonii, Actinomyces naeslundii, Fusobacterium nucleatum, and Porphyromonas gingivalis on rigid gas-permeable contact lenses. Biofilms, as well as planktonic cultures of these same bacterial species, were incubated under anaerobic conditions with a human oral epithelial cell line, OKF4, for up to 24h. Gro-1α, IL1α, IL-6, IL-8, TGFα, Fractalkine, MIP-1α, and IP-10 were shown to be produced in response to a range of the planktonic or biofilm forms of these species. P. gingivalis biofilms significantly inhibited the production of all of these cytokines and chemokines, except MIP-1α. Generally, the biofilms of all species inhibited Gro-1α, TGFα, and Fractalkine production, while F. nucleatum biofilms stimulated significant increases in IL-1α, IL-6, IL-8, and IP-10. A. naeslundii biofilms induced elevated levels of IL-6, IL-8 and IP-10. The oral streptococcal species in biofilms or planktonic forms were poor stimulants for any of these mediators from the epithelial cells. The results of these studies demonstrate that oral bacteria in biofilms elicit a substantially different profile of responses compared to planktonic bacteria of the same species. Moreover, certain oral species are highly stimulatory when in biofilms and interact with host cell receptors to trigger pathways of responses that appear quite divergent from individual bacteria. PMID:22266273

  5. Analysis of different approaches for evaluation of surface energy of microbial cells by contact angle goniometry.

    PubMed

    Sharma, P K; Rao, K Hanumantha

    2002-08-01

    Microbial adhesion on solid substrate is important in various fields of science. Mineral-microbe interactions alter the surface chemistry of the minerals and the adhesion of the bacterial cells to mineral surface is a prerequisite in several biobeneficiation processes. Apart from the surface charge and hydrophobic or hydrophilic character of the bacterial cells, the surface energy is a very important parameter influencing their adhesion on solid surfaces. There were many thermodynamic approaches in the literature to evaluate the cells surface energy. Although contact angle measurements with different liquids with known surface tension forms the basis in the calculation of the value of surface energy of solids, the results are different depending on the approach followed. In the present study, the surface energy of 140 bacterial and seven yeast cell surfaces has been studied following Fowkes, Equation of state, Geometric mean and Lifshitz-van der Waals acid-base (LW-AB) approaches. Two independent issues were addressed separately in our analysis. At first, the surface energy and the different components of the surface energy for microbial cells surface are examined. Secondly, the different approaches are evaluated for their internal consistency, similarities and dissimilarities. The Lifshitz-van der Waals component of surface energy for most of the microbial cells is realised to be approximately 40 mJ/m2 +/-10%. Equation of state and Geometric mean approaches do not possess any internal consistency and yield different results. The internal consistency of the LW-AB approach could be checked only by varying the apolar liquid and it evaluates coherent surface energy parameters by doing so. The electron-donor surface energy component remains exactly the same with the change of apolar liquid. This parameter could differentiate between the Gram-positive and Gram-negative bacterial cells. Gram-negative bacterial cells having higher electron-donor parameter had lower

  6. Bioinspired Nanosucker Array for Enhancing Bioelectricity Generation in Microbial Fuel Cells.

    PubMed

    Wang, Wei; You, Shijie; Gong, Xiaobo; Qi, Dianpeng; Chandran, Bevita K; Bi, Lanpo; Cui, Fuyi; Chen, Xiaodong

    2016-01-13

    A bioinspired active anode with a suction effect is demonstrated for microbial fuel cells by constructing polypyrrole (PPy) nanotubular arrays on carbon textiles. The oxygen in the inner space of the nanosucker can be depleted by micro-organisms with the capability of facul-tative respiration, forming a vacuum, which then activates the electrode to draw the microorganism by suction and thus improve the bioelectricity generation. PMID:26550771

  7. Biofuel Cells Select for Microbial Consortia That Self-Mediate Electron Transfer

    PubMed Central

    Rabaey, Korneel; Boon, Nico; Siciliano, Steven D.; Verhaege, Marc; Verstraete, Willy

    2004-01-01

    Microbial fuel cells hold great promise as a sustainable biotechnological solution to future energy needs. Current efforts to improve the efficiency of such fuel cells are limited by the lack of knowledge about the microbial ecology of these systems. The purposes of this study were (i) to elucidate whether a bacterial community, either suspended or attached to an electrode, can evolve in a microbial fuel cell to bring about higher power output, and (ii) to identify species responsible for the electricity generation. Enrichment by repeated transfer of a bacterial consortium harvested from the anode compartment of a biofuel cell in which glucose was used increased the output from an initial level of 0.6 W m−2 of electrode surface to a maximal level of 4.31 W m−2 (664 mV, 30.9 mA) when plain graphite electrodes were used. This result was obtained with an average loading rate of 1 g of glucose liter−1 day−1 and corresponded to 81% efficiency for electron transfer from glucose to electricity. Cyclic voltammetry indicated that the enhanced microbial consortium had either membrane-bound or excreted redox components that were not initially detected in the community. Dominant species of the enhanced culture were identified by denaturing gradient gel electrophoresis and culturing. The community consisted mainly of facultative anaerobic bacteria, such as Alcaligenes faecalis and Enterococcus gallinarum, which are capable of hydrogen production. Pseudomonas aeruginosa and other Pseudomonas species were also isolated. For several isolates, electrochemical activity was mainly due to excreted redox mediators, and one of these mediators, pyocyanin produced by P. aeruginosa, could be characterized. Overall, the enrichment procedure, irrespective of whether only attached or suspended bacteria were examined, selected for organisms capable of mediating the electron transfer either by direct bacterial transfer or by excretion of redox components. PMID:15345423

  8. Sequencing Single Cell Microbial Genomes with Microfluidic Amplifications Tools (MICW - Metagenomics Informatics Challenges Workshop: 10K Genomes at a Time)

    SciTech Connect

    Quake, Steve

    2011-10-12

    Stanford University's Steve Quake on "Sequencing Single Cell Microbial Genomes with Microfluidic Amplification Tools" at the Metagenomics Informatics Challenges Workshop held at the DOE JGI on October 12-13, 2011.

  9. Sequencing Single Cell Microbial Genomes with Microfluidic Amplifications Tools (MICW - Metagenomics Informatics Challenges Workshop: 10K Genomes at a Time)

    ScienceCinema

    Quake, Steve [University of Stanford

    2013-01-22

    Stanford University's Steve Quake on "Sequencing Single Cell Microbial Genomes with Microfluidic Amplification Tools" at the Metagenomics Informatics Challenges Workshop held at the DOE JGI on October 12-13, 2011.

  10. Overexpression of peptide-methionine sulfoxide reductase in Saccharomyces cerevisiae and human T cells provides them with high resistance to oxidative stress

    PubMed Central

    Moskovitz, Jackob; Flescher, Eliezer; Berlett, Barbara S.; Azare, Janeen; Poston, J. Michael; Stadtman, Earl R.

    1998-01-01

    The yeast peptide-methionine sulfoxide reductase (MsrA) was overexpressed in a Saccharomyces cerevisiae null mutant of msrA by using a high-copy plasmid harboring the msrA gene and its promoter. The resulting strain had about 25-fold higher MsrA activity than its parent strain. When exposed to either hydrogen peroxide, paraquat, or 2,2′-azobis-(2-amidinopropane) dihydrochloride treatment, the MsrA overexpressed strain grew better, had lower free and protein-bound methionine sulfoxide and had a better survival rate under these conditions than did the msrA mutant and its parent strain. Substitution of methionine with methionine sulfoxide in a medium lacking hydrogen peroxide had little effect on the growth pattern, which suggests that the oxidation of free methionine in the growth medium was not the main cause of growth inhibition of the msrA mutant. Ultraviolet A radiation did not result in obvious differences in survival rates among the three strains. An enhanced resistance to hydrogen peroxide treatment was shown in human T lymphocyte cells (Molt-4) that were stably transfected with the bovine msrA and exposed to hydrogen peroxide. The survival rate of the transfected strain was much better than its parent strain when grown in the presence of hydrogen peroxide. These results support the proposition that the msrA gene is involved in the resistance of yeast and mammalian cells to oxidative stress. PMID:9826655

  11. Multicopy Fzf1 (Sul1) Suppresses the Sulfite Sensitivity but Not the Glucose Derepression or Aberrant Cell Morphology of a Grr1 Mutant of Saccharomyces Cerevisiae

    PubMed Central

    Avram, D.; Bakalinsky, A. T.

    1996-01-01

    An ssu2 mutation in Sacccharomyces cerevisiae, previously shown to cause sulfite sensitivity, was found to be allelic to GRR1, a gene previously implicated in glucose repression. The suppressor rgt1, which suppresses the growth defects of grr1 strains on glucose, did not fully suppress the sensitivity on glucose or nonglucose carbon sources, indicating that it is not strictly linked to a defect in glucose metabolism. Because the Cln1 protein was previously shown to be elevated in grr1 mutants, the effect of CLN1 overexpression on sulfite sensitivity was investigated. Overexpression in GRR1 cells resulted in sulfite sensitivity, suggesting a connection between CLN1 and sulfite metabolism. Multicopy FZF1, a putative transcription factor, was found to suppress the sulfite sensitive phenotype of grr1 strains, but not the glucose derepression or aberrant cell morphology. Multicopy FZF1 was also found to suppress the sensitivity of a number of other unrelated sulfite-sensitive mutants, but not that of ssu1 or met20, implying that FZF1 may act through Ssu1p and Met20p. Disruption of FZF1 resulted in sulfite sensitivity when the construct was introduced in single copy at the FZF1 locus in a GRR1 strain, providing evidence that FZF1 is involved in sulfite metabolism. PMID:8889516

  12. Beta-glucan-depleted, glycopeptide-rich extracts from Brewer's and Baker's yeast (Saccharomyces cerevisiae) lower interferon-gamma production by stimulated human blood cells in vitro.

    PubMed

    Williams, Roderick; Dias, Daniel A; Jayasinghe, Nirupama; Roessner, Ute; Bennett, Louise E

    2016-04-15

    Regulation of the human immune system requires controlled pro- and anti-inflammatory responses for host defence against infection and disease states. Yeasts (Saccharomyces cerevisiae), as used in brewing and baking, are mostly known for ability to stimulate the human immune-system predominantly reflecting the pro-inflammatory cell wall β-glucans. However, in this study, using food-compatible processing methods, glycopeptide-enriched and β-glucan-depleted products were each prepared from Brewer's and Baker's yeasts, which suppressed production of interferon-γ (IFN-γ) in human whole blood cell assay, signifying that anti-inflammatory factors are also present in yeast. Anti-inflammatory bioactivities of products prepared from Brewer's and Baker's yeast were compared with the commercial yeast product, Epicor®. While unfractionated Epicor was inactive, the C18 resin-binding fractions of Brewer's and Baker's yeast products and Epicor dose-dependently lowered IFN-γ, demonstrating that Epicor also contained both pro-inflammatory (β-glucans) and anti-inflammatory components. Anti-inflammatory activity was attributed to C18 resin-binding species glyco-peptides in Epicor and experimental yeast products. This study demonstrated that pro- and anti-inflammatory factors could be resolved and enriched in yeasts by suitable processing, with potential to improve specific activities. PMID:26617014

  13. Activity of mitochondrially synthesized reporter proteins is lower than that of imported proteins and is increased by lowering cAMP in glucose-grown Saccharomyces cerevisiae cells.

    PubMed Central

    Demlow, Christina M; Fox, Thomas D

    2003-01-01

    We selected for increased phenotypic expression of a synthetic cox2::arg8m-G66S reporter gene inserted into Saccharomyces cerevisiae mtDNA in place of COX2. Recessive mutations in ras2 and cyr1, as well as elevated dosage of PDE2, allowed cox2::arg8m-G66S to support Arg prototrophy. Each of these genetic alterations should decrease cellular cAMP levels. The resulting signal was transduced through redundant action of the three cAMP-dependent protein kinases, TPK1, TPK2, and TPK3. ras2 had little or no effect on the level of wild-type Arg8p encoded by cox2::ARG8m, but did increase Arg8p activity, as judged by growth phenotype. ras2 also caused increased fluorescence in cells carrying the synthetic cox3::GFPm reporter in mtDNA, but had little effect on the steady-state level of GFP polypeptide detected immunologically. Thus, decreased cAMP levels did not affect the synthesis of mitochondrially coded protein reporters in glucose-grown cells, but rather elevated activities in the matrix that promote efficient folding. Furthermore, we show that when Arg8p is synthesized in the cytoplasm and imported into mitochondria, it has greater activity than when it is synthesized in the matrix. Thus, mitochondrially synthesized proteins may not have the same access to matrix chaperones as cytoplasmically synthesized proteins emerging from the import apparatus. PMID:14668357

  14. Submersible microbial fuel cell sensor for monitoring microbial activity and BOD in groundwater: focusing on impact of anodic biofilm on sensor applicability.

    PubMed

    Zhang, Yifeng; Angelidaki, Irini

    2011-10-01

    A sensor, based on a submersible microbial fuel cell (SUMFC), was developed for in situ monitoring of microbial activity and biochemical oxygen demand (BOD) in groundwater. Presence or absence of a biofilm on the anode was a decisive factor for the applicability of the sensor. Fresh anode was required for application of the sensor for microbial activity measurement, while biofilm-colonized anode was needed for utilizing the sensor for BOD content measurement. The current density of SUMFC sensor equipped with a biofilm-colonized anode showed linear relationship with BOD content, to up to 250 mg/L (∼233 ± 1 mA/m(2)), with a response time of <0.67 h. This sensor could, however, not measure microbial activity, as indicated by the indifferent current produced at varying active microorganisms concentration, which was expressed as microbial adenosine-triphosphate (ATP) concentration. On the contrary, the current density (0.6 ± 0.1 to 12.4 ± 0.1 mA/m(2)) of the SUMFC sensor equipped with a fresh anode showed linear relationship, with active microorganism concentrations from 0 to 6.52 nmol-ATP/L, while no correlation between the current and BOD was observed. It was found that temperature, pH, conductivity, and inorganic solid content were significantly affecting the sensitivity of the sensor. Lastly, the sensor was tested with real contaminated groundwater, where the microbial activity and BOD content could be detected in <3.1 h. The microbial activity and BOD concentration measured by SUMFC sensor fitted well with the one measured by the standard methods, with deviations ranging from 15% to 22% and 6% to 16%, respectively. The SUMFC sensor provides a new way for in situ and quantitative monitoring contaminants content and biological activity during bioremediation process in variety of anoxic aquifers. PMID:21557205

  15. Oral epithelial cell responses to multispecies microbial biofilms.

    PubMed

    Peyyala, R; Kirakodu, S S; Novak, K F; Ebersole, J L

    2013-03-01

    This report describes the use of a novel model of multispecies biofilms to stimulate profiles of cytokines/chemokines from oral epithelial cells that contribute to local inflammation in the periodontium. Streptococcus gordonii (Sg)/S. oralis (So)/S. sanguinis (Ss) and Sg/Fusobacterium nucleatum (Fn)/Porphyromonas gingivalis (Pg) biofilms elicited significantly elevated levels of IL-1α and showed synergistic stimulatory activity compared with an additive effect of the 3 individual bacteria. Only the Sg/Actinomyces naeslundii (An)/Fn multispecies biofilms elicited IL-6 levels above those of control. IL-8 was a primary response to the Sg/An/Fn biofilms, albeit the level was not enhanced compared with a predicted composite level from the monospecies challenges. These results represent some of the first data documenting alterations in profiles of oral epithelial cell responses to multispecies biofilms. PMID:23300185

  16. Cell immobilization for microbial production of 1,3-propanediol.

    PubMed

    Gungormusler-Yilmaz, Mine; Cicek, Nazim; Levin, David B; Azbar, Nuri

    2016-06-01

    Cell and enzyme immobilization are often used for industrial production of high-value products. In recent years, immobilization techniques have been applied to the production of value-added chemicals such as 1,3-Propanediol (1,3-PDO). Biotechnological fermentation is an attractive alternative to current 1,3-PDO production methods, which are primarily thermochemical processes, as it generates high volumetric yields of 1,3-PDO, is a much less energy intensive process, and generates lower amounts of environmental organic pollutants. Although several approaches including: batch, fed-batch, continuous-feed and two-step continuous-feed were tested in suspended systems, it has been well demonstrated that cell immobilization techniques can significantly enhance 1,3-PDO production and allow robust continuous production in smaller bioreactors. This review covers various immobilization methods and their application for 1,3-PDO production. PMID:25600463

  17. Gate control: guard cell regulation by microbial stress.

    PubMed

    McLachlan, Deirdre H; Kopischke, Michaela; Robatzek, Silke

    2014-09-01

    Terrestrial plants rely on stomata, small pores in the leaf surface, for photosynthetic gas exchange and transpiration of water. The stomata, formed by a pair of guard cells, dynamically increase and decrease their volume to control the pore size in response to environmental cues. Stresses can trigger similar or opposing movements: for example, drought induces closure of stomata, whereas many pathogens exploit stomata and cause them to open to facilitate entry into plant tissues. The latter is an active process as stomatal closure is part of the plant's immune response. Stomatal research has contributed much to clarify the signalling pathways of abiotic stress, but guard cell signalling in response to microbes is a relatively new area of research. In this article, we discuss present knowledge of stomatal regulation in response to microbes and highlight common points of convergence, and differences, compared to stomatal regulation by abiotic stresses. We also expand on the mechanisms by which pathogens manipulate these processes to promote disease, for example by delivering effectors to inhibit closure or trigger opening of stomata. The study of pathogen effectors in stomatal manipulation will aid our understanding of guard cell signalling. PMID:25040778

  18. Rapid and high-throughput construction of microbial cell-factories with regulatory noncoding RNAs.

    PubMed

    Chaudhary, Amit Kumar; Na, Dokyun; Lee, Eun Yeol

    2015-11-01

    Due to global crises such as pollution and depletion of fossil fuels, sustainable technologies based on microbial cell-factories have been garnering great interest as an alternative to chemical factories. The development of microbial cell-factories is imperative in cutting down the overall manufacturing cost. Thus, diverse metabolic engineering strategies and engineering tools have been established to obtain a preferred genotype and phenotype displaying superior productivity. However, these tools are limited to only a handful of genes with permanent modification of a genome and significant labor costs, and this is one of the bottlenecks associated with biofactory construction. Therefore, a groundbreaking rapid and high-throughput engineering tool is needed for efficient construction of microbial cell-factories. During the last decade, copious small noncoding RNAs (ncRNAs) have been discovered in bacteria. These are involved in substantial regulatory roles like transcriptional and post-transcriptional gene regulation by modulating mRNA elongation, stability, or translational efficiency. Because of their vulnerability, ncRNAs can be used as another layer of conditional control over gene expression without modifying chromosomal sequences, and hence would be a promising high-throughput tool for metabolic engineering. Here, we review successful design principles and applications of ncRNAs for high-throughput metabolic engineering or physiological studies of diverse industrially important microorganisms. PMID:26027891

  19. In situ bioremediation of trichloroethylene-contaminated water by a resting-cell methanotrophic microbial filter

    SciTech Connect

    Taylor, R T; Duba, A G; Durham, W B; Hanna, M L; Jackson, K J; Jovanovich, M C; Knapp, R B; Knezovich, J P; Shah, N N; Shonnard, D R; Wijesinghe, A M

    1992-10-01

    The Lawrence Livermore National Laboratory is testing and developing an in situ microbial filter technology for remediating migrating subsurface plumes contaminated with low concentrations of trichloroethylene (TCE). Their current focus is the establishment of a replenishable bioactive zone (catalytic filter) along expanding plume boundaries by the Injection of a representative methanotrophic bacterium, Methylosinus trichosporium OB3b. We have successfully demonstrated this microbial filter strategy using emplaced, attached resting cells (no methane additions) in a 1.1-m flow-through test bed loaded with water-saturated sand. Two separate 24 h pulses of TCE (109 ppb and 85 ppb), one week apart, were pumped through the system at a flow velocity of 1.5 cm/h; no TCE (<0.5 ppb) was detected on the downstream side of the microbial filter. Subsequent excavation of the wet sand confirmed the existence of a TCE-bioactive zone 19 days after it had been created. An enhanced longevity of the cellular, soluble-form methane monooxygenase produced by this methanotroph Is a result of our laboratory bioreactor culturing conditions. Additional experiments with cells in sealed vials and emplaced in the 1.1-m test bed yielded a high resting-cell finite TCE biotransformation capacity of [approximately] 0.25 mg per mg of bacteria; this is suitable for a planned sand-filled trench field demonstration at a Lawrence Livermore National Laboratory site.

  20. Photosynthetic microbial desalination cells (PMDCs) for clean energy, water and biomass production.

    PubMed

    Kokabian, Bahareh; Gude, Veera Gnaneswar

    2013-12-01

    Current microbial desalination cell (MDC) performances are evaluated with chemical catalysts such as ferricyanide, platinum catalyzed air-cathodes or aerated cathodes. All of these methods improve power generation potential in MDCs, however, they are not preferable for large scale applications due to cost, energy and environmental toxicity issues. In this study, performance of microbial desalination cells with an air cathode and an algae biocathode (Photosynthetic MDC - PMDC) were evaluated, both under passive conditions (no mechanical aeration or mixing). The results indicate that passive algae biocathodes perform better than air cathodes and enhance COD removal and utilize treated wastewater as the growth medium to obtain valuable biomass for high value bioproducts. Maximum power densities of 84 mW m(-3) (anode volume) or 151 mW m(-3) (biocathode volume) and a desalination rate of 40% were measured with 0.9 : 1 : 0.5 volumetric ratios of anode, desalination and algae biocathode chambers respectively. This first proof-of-concept study proves that the passive mechanisms can be beneficial in enhancing the sustainability of microbial desalination cells. PMID:24154718

  1. Graphene/Nickel Nanofiber Hybrids for Catalytic and Microbial Fuel Cell Applications.

    PubMed

    Kartick, B; Srivastava, S K; Chandra, A

    2016-01-01

    The present work deals with the preparation of fcc-nickel (Ni) nanofibers and Graphene/Ni hybrids. The analysis of XRD, FTIR and Raman data confirms the formation of pure Ni, graphite oxide, reduced graphene and their hybrids. SEM micrographs clearly show the decoration of Ni nanofibers on the graphene flakes. The synthesized Ni-based hybrid systems can have applications in areas ranging from: magnetism, catalysis to microbial fuel cells. The vibrating sample magnetometer (VSM) investigations reveal that the hybrid structure shows hysteresis loop similar to that expected from a superparamagnetic system. When the same hybrid structure is used as catalyst for decolourization of 4-nitrophenol, the maximum rate constant of 0.1 min⁻¹ can be obtained. The catalytic activity also shows dependence on the loading concentration of Ni. The enhanced performance can be explained on the basis of synergistic effect between graphene and nickel nanofibers. The developed hybrids have also been applied as catalyst for cathode in microbial fuel cells and the studies showed improved power density compared to conventional microbial fuel cells (MFCs). PMID:27398458

  2. Ribosomal acidic phosphoproteins P1 and P2 are not required for cell viability but regulate the pattern of protein expression in Saccharomyces cerevisiae.

    PubMed Central

    Remacha, M; Jimenez-Diaz, A; Bermejo, B; Rodriguez-Gabriel, M A; Guarinos, E; Ballesta, J P

    1995-01-01

    Saccharomyces cerevisiae strains with either three inactivated genes (triple disruptants) or four inactivated genes (quadruple disruptants) encoding the four acidic ribosomal phosphoproteins, YP1 alpha, YP1 beta, YP2 alpha, and YP2 beta, present in this species have been obtained. Ribosomes from the triple disruptants and, obviously, those from the quadruple strain do not have bound P proteins. All disrupted strains are viable; however, they show a cold-sensitive phenotype, growing very poorly at 23 degrees C. Cell extracts from the quadruple-disruptant strain are about 30% as active as the control in protein synthesis assays and are stimulated by the addition of free acidic P proteins. Strains lacking acidic proteins do not have a higher suppressor activity than the parental strains, and cell extracts derived from the quadruple disruptant do not show a higher degree of misreading, indicating that the absence of acidic proteins does not affect the accuracy of the ribosomes. However, the patterns of protein expressed in the cells as well as in the cell-free protein system are affected by the absence of P proteins from the particles; a wild-type pattern is restored upon addition of exogenous P proteins to the cell extract. In addition, strains carrying P-protein-deficient ribosomes are unable to sporulate but recover this capacity upon transformation with one of the missing genes. These results indicate that acidic proteins are not an absolute requirement for protein synthesis but regulate the activity of the 60S subunit, affecting the translation of certain mRNAs differently. PMID:7651393

  3. Biotransformation of malachite green by Saccharomyces cerevisiae MTCC 463.

    PubMed

    Jadhav, J P; Govindwar, S P

    2006-03-01

    In recent years, use of microbial biomass for decolourization of textile industry wastewater is becoming a promising alternative in which some bacteria and fungi are used to replace present treatment processes. Saccharomyces cerevisiae MTCC 463 decolourized the triphenylmethane dyes (malachite green, cotton blue, methyl violet and crystal violet) by biosorption, showing different decolourization patterns. However, malachite green decolourized by biosorption at the initial stage and further biodegradation occurred, about 85% in plain distilled water within 7 h, and about 95.5% in 5% glucose medium within 4 h, under aerobic conditions and at room temperature. Decolourization of malachite green depends on various conditions, such as concentration of dye, concentration of cells, composition of medium and agitation. HPLC, UV-VIS, FTIR and TLC analysis of samples extracted with ethyl acetate from decolourized culture flasks confirmed the biodegradation of malachite green into several metabolites. A study of the enzymes responsible for the biodegradation of malachite green in the control and cells obtained after decolourization showed the activities of laccase, lignin peroxidase, NADH-DCIP reductase, malachite green reductase and aminopyrine N-demethylase in control cells. A significant increase in the activities of NADH-DCIP reductase and MG reductase was observed in the cells obtained after decolourization, indicating a major involvement of reductases in malachite green degradation. PMID:16544273

  4. Biosorption of heavy metals by Saccharomyces cerevisiae.

    PubMed

    Volesky, B; May-Phillips, H A

    1995-01-01

    Abundant and common yeast biomass has been examined for its capacity to sequester heavy metals from dilute aqueous solutions. Live and non-living biomass of Saccharomyces cerevisiae differs in the uptake of uranium, zinc and copper at the optimum pH 4-5. Culture growth conditions can influence the biosorbent metal uptake capacity which normally was: living and non-living brewer's yeast: U > Zn > Cd > Cu; non-living baker's yeast: Zn > (Cd) > U > Cu; living baker's yeast: Zn > Cu approximately (Cd) > U. Non-living brewer's yeast biomass accumulated 0.58 mmol U/g. The best biosorbent of zinc was non-living baker's yeast (approximately 0.56 mmol Zn/g). Dead cells of S. cerevisiae removed approximately 40% more uranium or zinc than the corresponding live cultures. Biosorption of uranium by S. cerevisiae was a rapid process reaching 60% of the final uptake value within the first 15 min of contact. Its deposition differing from that of other heavy metals more associated with the cell wall, uranium was deposited as fine needle-like crystals both on the inside and outside of the S. cerevisiae cells. PMID:7765919

  5. Sporulation in the Budding Yeast Saccharomyces cerevisiae

    PubMed Central

    Neiman, Aaron M.

    2011-01-01

    In response to nitrogen starvation in the presence of a poor carbon source, diploid cells of the yeast Saccharomyces cerevisiae undergo meiosis and package the haploid nuclei produced in meiosis into spores. The formation of spores requires an unusual cell division event in which daughter cells are formed within the cytoplasm of the mother cell. This process involves the de novo generation of two different cellular structures: novel membrane compartments within the cell cytoplasm that give rise to the spore plasma membrane and an extensive spore wall that protects the spore from environmental insults. This article summarizes what is known about the molecular mechanisms controlling spore assembly with particular attention to how constitutive cellular functions are modified to create novel behaviors during this developmental process. Key regulatory points on the sporulation pathway are also discussed as well as the possible role of sporulation in the natural ecology of S. cerevisiae. PMID:22084423

  6. Microbial cell wall agents as an occupational hazard

    SciTech Connect

    Sigsgaard, T. . E-mail: ts@mil.au.dk; Bonefeld-Jorgensen, E.C.; Hoffmann, H.J.; Bonlokke, J.; Krueger, T.

    2005-09-01

    Organic dusts cause inflammatory reactions in the tissues exposed. The lung and the cells lining the surface of the respiratory tract are a primary target. Many receptors have been shown to react specifically on the presence of microorganisms that are ubiquitous elements in organic dusts. There is a great variability in the individual response to organic dusts. Almost 50% of Caucasians are hyporesponders to LPS exposure, and people with alpha-1-antitrypsin deficiency are hyperresponsive to organic dust exposure. The diseases resulting from organic dust exposures include asthma, allergy, hypersensitivity pneumonitis and toxic pneumonitis (organic dust toxic syndrome).This paper deals with inflammation and the subsequent mechanism of disease as it is encountered in industries with these exposures. Toxicological studies including human experimental exposures and ex vivo studies of cells are described. Cellular reactions are mediated through the attachment of, e.g. LPS and {beta} (1,3)-D-glucan to lipopolysaccharide binding protein, CD14 and Toll-like receptors. The relation between protein release and the gene activation is described. Furthermore, studies of the individual susceptibility will be reviewed.

  7. High rate copper and energy recovery in microbial fuel cells

    PubMed Central

    Rodenas Motos, Pau; ter Heijne, Annemiek; van der Weijden, Renata; Saakes, Michel; Buisman, Cees J. N.; Sleutels, Tom H. J. A.

    2015-01-01

    Bioelectrochemical systems (BESs) are a novel, promising technology for the recovery of metals. The prerequisite for upscaling from laboratory to industrial size is that high current and high power densities can be produced. In this study we report the recovery of copper from a copper sulfate stream (2 g L-1 Cu2+) using a laboratory scale BES at high rate. To achieve this, we used a novel cell configuration to reduce the internal voltage losses of the system. At the anode, electroactive microorganisms produce electrons at the surface of an electrode, which generates a stable cell voltage of 485 mV when combined with a cathode where copper is reduced. In this system, a maximum current density of 23 A m-2 in combination with a power density of 5.5 W m-2 was produced. XRD analysis confirmed 99% purity in copper of copper deposited onto cathode surface. Analysis of voltage losses showed that at the highest current, most voltage losses occurred at the cathode, and membrane, while anode losses had the lowest contribution to the total voltage loss. These results encourage further development of BESs for bioelectrochemical metal recovery. PMID:26150802

  8. Signature gene expressions of cell wall integrity pathway concur with tolerance response of industrial yeast Saccharomyces cerevisiae against biomass pretreatment inhibitors

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Traditional industrial ethanologenic yeast Saccharomyces cerevisiae has a robust performance under various environmental conditions and can be served as a candidate for the next-generation biocatalyst development for advanced biofuels production using lignocellulose mateials. Overcoming toxic compou...

  9. Immobilization of anode-attached microbes in a microbial fuel cell

    PubMed Central

    2012-01-01

    Current-generating (exoelectrogenic) bacteria in bioelectrochemical systems (BESs) may not be culturable using standard in vitro agar-plating techniques, making isolation of new microbes a challenge. More in vivo like conditions are needed where bacteria can be grown and directly isolated on an electrode. While colonies can be developed from single cells on an electrode, the cells must be immobilized after being placed on the surface. Here we present a proof-of-concept immobilization approach that allows exoelectrogenic activity of cells on an electrode based on applying a layer of latex to hold bacteria on surfaces. The effectiveness of this procedure to immobilize particles was first demonstrated using fluorescent microspheres as bacterial analogs. The latex coating was then shown to not substantially affect the exoelectrogenic activity of well-developed anode biofilms in two different systems. A single layer of airbrushed coating did not reduce the voltage produced by a biofilm in a microbial fuel cell (MFC), and more easily applied dip-and-blot coating reduced voltage by only 11% in a microbial electrolysis cell (MEC). This latex immobilization procedure will enable future testing of single cells for exoelectrogenic activity on electrodes in BESs. PMID:22214379

  10. Use of Surface Enhanced Blocking (SEB) Electrodes for Microbial Cell Lysis in Flow-Through Devices

    PubMed Central

    Talebpour, Abdossamad; Maaskant, Robert; Khine, Aye Aye; Alavie, Tino

    2014-01-01

    By simultaneously subjecting microbial cells to high amplitude pulsed electric fields and flash heating of the cell suspension fluid, effective release of intracellular contents was achieved. The synergistic effect of the applied electric field and elevated temperature on cell lysis in a flow-through device was demonstrated for Gram-negative and Gram-positive bacteria, and Mycobacterium species. The resulting lysate is suitable for downstream nucleic acid amplification and detection without requiring further preparation. The lysis chamber employs surface enhanced blocking electrodes which possess an etched micro-structured surface and a thin layer of dielectric metal oxide which provides a large effective area and blocks transmission of electrical current. The surface enhanced blocking electrodes enable simultaneous suppression of the rapid onset of electric field screening in the bulk of the cell suspension medium and avoidance of undesired electrochemical processes at the electrode-electrolyte interface. In addition the blocking layer ensures the robustness of the cell lysis device in applications involving prolonged flow-through processing of the microbial cells. PMID:25033080

  11. Modification of a High-Throughput Automatic Microbial Cell Enumeration System for Shipboard Analyses.

    PubMed

    Bennke, Christin M; Reintjes, Greta; Schattenhofer, Martha; Ellrott, Andreas; Wulf, Jörg; Zeder, Michael; Fuchs, Bernhard M

    2016-06-01

    In the age of ever-increasing "-omics" studies, the accurate and statistically robust determination of microbial cell numbers within often-complex samples remains a key task in microbial ecology. Microscopic quantification is still the only method to enumerate specific subgroups of microbial clades within complex communities by, for example, fluorescence in situ hybridization (FISH). In this study, we improved an existing automatic image acquisition and cell enumeration system and adapted it for usage at high seas on board an oceanographic research ship. The system was evaluated by testing settings such as minimal pixel area and image exposure times ashore under stable laboratory conditions before being brought on board and tested under various wind and wave conditions. The system was robust enough to produce high-quality images even with ship heaves of up to 3 m and pitch and roll angles of up to 6.3°. On board the research ship, on average, 25% of the images acquired from plankton samples on filter membranes could be used for cell enumeration. Automated enumeration was highly correlated with manual counts (r(2) > 0.9). Even the smallest of microbial cells in the open ocean, members of the alphaproteobacterial SAR11 clade, could be confidently detected and enumerated. The automated image acquisition and cell enumeration system developed here enables an accurate and reproducible determination of microbial cell counts in planktonic samples and allows insight into the abundance and distribution of specific microorganisms already on board within a few hours.IMPORTANCE In this research article, we report on a new system and software pipeline, which allows for an easy and quick image acquisition and the subsequent enumeration of cells in the acquired images. We put this pipeline through vigorous testing and compared it to manual microscopy counts of microbial cells on membrane filters. Furthermore, we tested this system at sea on board a marine research vessel and

  12. Use of Saccharum spontaneum (wild sugarcane) as biomaterial for cell immobilization and modulated ethanol production by thermotolerant Saccharomyces cerevisiae VS3.

    PubMed

    Chandel, Anuj K; Narasu, M Lakshmi; Chandrasekhar, G; Manikyam, A; Rao, L Venkateswar

    2009-04-01

    Saccharum spontaneum is a wasteland weed consists of 45.10+/-0.35% cellulose and 22.75+/-0.28% of hemicellulose on dry solid (DS) basis. Aqueous ammonia delignified S. spontaneum yielded total reducing sugars, 53.91+/-0.44 g/L (539.10+/-0.55 mg/g of substrate) with a hydrolytic efficiency of 77.85+/-0.45%. The enzymes required for hydrolysis were prepared from culture supernatants of Aspergillus oryzae MTCC 1846. A maximum of 0.85+/-0.07 IU/mL of filter paperase (FPase), 1.25+/-0.04 IU/mL of carboxy methyl cellulase (CMCase) and 55.56+/-0.52 IU/mL of xylanase activity was obtained after 7 days of incubation at 28+/-0.5 degrees C using delignified S. spontaneum as carbon source under submerged fermentation conditions. Enzymatic hydrolysate of S. spontaneum was then tested for ethanol production under batch and repeated batch production system using "in-situ" entrapped Saccharomyces cerevisiae VS3 cells in S. spontaneum stalks (1 cm x 1 cm) size. Immobilization was confirmed by the scanning electron microscopy (SEM). Batch fermentation of VS3 free cells and immobilized cells showed ethanol production, 19.45+/-0.55 g/L (yield, 0.410+/-0.010 g/g) and 21.66+/-0.62 g/L (yield, 0.434+/-0.021 g/g), respectively. Immobilized VS3 cells showed maximum ethanol production (22.85+/-0.44 g/L, yield, 0.45+/-0.04 g/g) up to 8th cycle during repeated batch fermentation followed by a gradual reduction in subsequent cycles of fermentation. PMID:19114303

  13. Cell abundance and microbial community composition along a complete oil sand mining and reclamation process

    NASA Astrophysics Data System (ADS)

    Lappé, M.; Schneider, B.; Kallmeyer, J.

    2012-12-01

    Hydrocarbons constitute an important energy source for microbes but can also be of environmental concern. Microbial activity causes hydrocarbon degradation and thereby loss of economical value, but also helps to remove hydrocarbons from the environment. The present study characterizes the abundance of microbes along the oil sand mining process in Alberta, Canada, as a first approach to assess the impact of mining and oil extraction on the microbial population. After mining the oil is extracted from the sediment by a hot-water extraction (50-60°C), resulting in three major fractions: crude oil, tailings sand and fine tailings. The tailings sand is used as substratum for newly developing soils on the reclamation areas. The very liquid fine tailings still have a TOC content of about 4.3% and are pumped into tailings ponds, where they need up to three decades to settle and solidify. After deposition, these mature fine tailings (MFTs) are enriched in organics (TOC content between 9.6 and 16.8%) and dredged out of the ponds and put on dumps for several years for dewatering. Finally they are brought out onto the reclamation sites and deposited below the sand layer. Cells were extracted from oily sediments according to the protocol of Lappé and Kallmeyer (2011), stained with SYBR Green I and counted by fluorescence microscopy. Cell abundance in the unprocessed oil sand is around 1.6 x 107 cells cm-3. After processing the fresh fine tailings still contain around 1.6 x 107 cells cm-3. Cell counts in the processed MFTs are 5.8 x 107 cells cm-3, whereas in the sand used as substratum for newly developing soils, they are twice as high (1.4 x 108). In root-bearing horizons, cell counts reach 1.1 x 109 cell cm-3. Cell numbers calculated from cultivation experiments are in the same range. Higher cell counts in the tailings sand are probably due to a higher nitrogen supply through the addition of a 35 cm top layer of a peat-mineral mix. In the sand nitrate concentrations are high

  14. Duty Cycling Influences Current Generation in Multi-Anode Environmental Microbial Fuel Cells

    SciTech Connect

    Gardel, EJ; Nielsen, ME; Grisdela, PT; Girguis, PR

    2012-05-01

    Improving microbial fuel cell (MFC) performance continues to be the subject of research, yet the role of operating conditions, specifically duty cycling, on MFC performance has been modestly addressed. We present a series of studies in which we use a 15-anode environmental MFC to explore how duty cycling (variations in the time an anode is connected) influences cumulative charge, current, and microbial composition. The data reveal particular switching intervals that result in the greatest time-normalized current. When disconnection times are sufficiently short, there is a striking decrease in current due to an increase in the overall electrode reaction resistance. This was observed over a number of whole cell potentials. Based on these results, we posit that replenishment of depleted electron donors within the biofilm and surrounding diffusion layer is necessary for maximum charge transfer, and that proton flux may be not limiting in the highly buffered aqueous phases that are common among environmental MFCs. Surprisingly, microbial diversity analyses found no discernible difference in gross community composition among duty cycling treatments, suggesting that duty cycling itself has little or no effect. Such duty cycling experiments are valuable in determining which factors govern performance of bioelectrochemical systems and might also be used to optimize field-deployed systems.

  15. Control of geometrical properties of carbon nanotube electrodes towards high-performance microbial fuel cells

    NASA Astrophysics Data System (ADS)

    Erbay, Celal; Pu, Xiong; Choi, Woongchul; Choi, Mi-Jin; Ryu, Yeontack; Hou, Huijie; Lin, Furong; de Figueiredo, Paul; Yu, Choongho; Han, Arum

    2015-04-01

    In microbial fuel cells (MFCs), physical and electrochemical interactions between microbes and electrode surfaces are critical to performance. Nanomaterial-based electrodes have shown promising performances, however their unique characteristics have not been fully utilized. The developed electrodes here consist of multi-wall carbon nanotubes (MWCNTs) directly grown in the radial direction from the wires of stainless steel (SS) meshes, providing extremely large three-dimensional surfaces while ensuring minimal ohmic loss between CNTs and SS meshes, fully utilizing the advantages of CNTs. Systematic studies on how different lengths, packing densities, and surface conditions of CNTs affect MFC power output revealed that long and loosely packed CNTs without any amorphous carbon show the highest power production performance. The power density of this anode is 7.4-fold higher compared to bare carbon cloth, which is the highest reported improvement for MFCs with nanomaterial-decorated electrodes. The results of this study offer great potential for advancing the development of microbial electrochemical systems by providing a highly efficient nanomaterial-based electrode that delivers large surface area, high electrochemical activity, and minimum ohmic loss, as well as provide design principles for next-generation nanomaterial-based electrodes that can be broadly applicable for highly efficient microbial electrochemical cells.

  16. Olive mill wastewater treatment in single-chamber air-cathode microbial fuel cells.

    PubMed

    Bermek, Hakan; Catal, Tunc; Akan, S Süha; Ulutaş, Mehmet Sefa; Kumru, Mert; Özgüven, Mine; Liu, Hong; Özçelik, Beraat; Akarsubaşı, Alper Tunga

    2014-04-01

    Olive mill wastewaters create significant environmental issues in olive-processing countries. One of the most hazardous groups of pollutants in these wastewaters is phenolic compounds. Here, olive mill wastewater was used as substrate and treated in single-chamber air-cathode microbial fuel cells. Olive mill wastewater yielded a maximum voltage of 381 mV on an external resistance of 1 kΩ. Notable decreases in the contents of 3,4-dihydroxybenzoic acid, tyrosol, gallic acid and p-coumaric acid were detected. Chemical oxygen demand removal rates were 65 % while removal of total phenolics by the process was lower (49 %). Microbial community analysis during the olive mill wastewater treating MFC has shown that both exoelectrogenic and phenol-degrading microorganisms have been enriched during the operation. Brevundimonas-, Sphingomonas- and Novosphingobium-related phylotypes were enriched on the anode biofilm, while Alphaproteobacteria and Bacteriodetes dominated the cathode biofilm. As one of the novel studies, it has been demonstrated that recalcitrant olive mill wastewaters could be treated and utilized for power generation in microbial fuel cells. PMID:24165748

  17. Microbial degradation of quinoline by immobilized cells of Burkholderia pickettii.

    PubMed

    Jianlong, Wang; Xiangchun, Quan; Liping, Han; Yi, Qian; Hegemann, Werner

    2002-05-01

    A quinoline-biodegrading microorganism was isolated from activated sludge of coke-oven wastewater treatment plant using quinoline as sole carbon and nitrogen source. It is a gram negative, rod-shaped and aerobic strain, which was identified as Burkholderia pickettii. The biodegradation of quinoline was carried out with this isolated strain. Analysis by high performance liquid chromatography and gas chromatography/mass spectrum (GC/MS) revealed that 2-hydroxyquinoline (2-OH-Q) was the first intermediate in the course of quinoline biodegradation. A novel immobilization carrier, that is, polyvinyl alcohol (PVA)-gauze hybrid carrier, was developed. The isolated strain was immobilized by two different immobilizing techniques and used for the quinolinerdegradation. It was found that biodegradation rate of quinoline by the microorganisms immobilized on PVA-gauze hybrid carrier was faster than that by the microorganisms immobilized in PVA gel beads. Kinetics of quinoline biodegradation by cells of Burkholderia pickettii immobilized on PVA-gauze hybrid carrier was investigated. The results demonstrate that quinoline degradation could be described by zero-order reaction rate equation when the initial quinoline concentration was in the range of 50-500 mg l(-1). PMID:12108721

  18. Microbial exposure alters HIV-1-induced mucosal CD4+ T cell death pathways Ex vivo

    PubMed Central

    2014-01-01

    Background Early HIV-1 infection causes massive CD4+ T cell death in the gut and translocation of bacteria into the circulation. However, the programmed cell death (PCD) pathways used by HIV-1 to kill CD4+ T cells in the gut, and the impact of microbial exposure on T cell loss, remain unclear. Understanding mucosal HIV-1 triggered PCD could be advanced by an ex vivo system involving lamina propria mononuclear cells (LPMCs). We therefore modeled the interactions of gut LPMCs, CCR5-tropic HIV-1 and a commensal gut bacterial species, Escherichia coli. In this Lamina Propria Aggregate Culture (LPAC) model, LPMCs were infected with HIV-1BaL by spinoculation and cultured in the presence or absence of heat killed E.coli. CD4+ T cell numbers derived from flow cytometry and viable cell counts were reported relative to mock infection. Viable cells were identified by viability dye exclusion (AqVi), and intracellular HIV-1 Gag p24 protein was used to identify infected cells. Annexin V and AqVi were used to identify apoptotic versus necrotic cells. Caspase-1 and Caspase-3 activities were blocked using specific inhibitors YVAD and DEVD, respectively. Results CD4+ T cell depletion following HIV-1 infection was reproducibly observed by 6 days post infection (dpi). Depletion at 6 dpi strongly correlated with infection frequency at 4 dpi, was significantly blocked by Efavirenz treatment, and was primarily driven by p24-negative cells that were predominantly necrotic. HIV-1 infection significantly induced CD4+ T-cell intrinsic Caspase-1 activity, whereas Caspase-1 inhibition, but not Caspase-3 inhibition, significantly blocked CD4+ T cell depletion. Exposure to E.coli enhanced HIV-1 infection and CD4+ T depletion, and significantly increased the number of apoptotic p24+ cells. Notably, CD4+ T cell depletion in the presence of E.coli was partially blocked by Caspase-3, but not by Caspase-1 inhibition. Conclusions In the LPAC model, HIV-1 induced Caspase-1 mediated pyroptosis in

  19. Effect of dissolved oxygen on nitrogen and phosphorus removal and electricity production in microbial fuel cell.

    PubMed

    Tao, Qinqin; Luo, Jingjing; Zhou, Juan; Zhou, Shaoqi; Liu, Guangli; Zhang, Renduo

    2014-07-01

    Performance of a two-chamber microbial fuel cell (MFC) was evaluated with the influence of cathodic dissolved oxygen (DO). The maximum voltage, coulombic efficiency and maximum power density outputs of MFC decreased from 521 to 303 mV, 52.48% to 23.09% and 530 to 178 mW/m(2) with cathodic DO declining. Furthermore, a great deal of total phosphorus (TP) was removed owing to chemical precipitation (about 80%) and microbial absorption (around 4-17%). COD was first removed in anode chamber (>70%) then in cathode chamber (<5%). Most of nitrogen was removed when the cathodic DO was at low levels. Chemical precipitates formed in cathode chamber were verified as phosphate, carbonate and hydroxyl compound with the aid of scanning electron microscope capable of energy dispersive spectroscopy (SEM-EDS), X-ray diffractometer (XRD) and Fourier transform infrared spectroscopy (FTIR). PMID:24880930

  20. Low-potential respirators support electricity production in microbial fuel cells.

    PubMed

    Grüning, André; Beecroft, Nelli J; Avignone-Rossa, Claudio

    2015-07-01

    In this paper, we analyse how electric power production in microbial fuel cells (MFCs) depends on the composition of the anodic biofilm in terms of metabolic capabilities of identified sets of species. MFCs are a promising technology for organic waste treatment and sustainable bioelectricity production. Inoculated with natural communities, they present a complex microbial ecosystem with syntrophic interactions between microbes with different metabolic capabilities. Our results demonstrate that low-potential anaerobic respirators--that is those that are able to use terminal electron acceptors with a low redox potential--are important for good power production. Our results also confirm that community metabolism in MFCs with natural inoculum and fermentable feedstock is a two-stage system with fermentation followed by anode respiration. PMID:25388758

  1. Organic acids from lignocellulose: Candida lignohabitans as a new microbial cell factory.

    PubMed

    Bellasio, Martina; Mattanovich, Diethard; Sauer, Michael; Marx, Hans

    2015-05-01

    Biorefinery applications require microbial cell factories for the conversion of various sugars derived from lignocellulosic material into value-added chemicals. Here, the capabilities of the yeast Candida lignohabitans to utilize a range of such sugars is characterized. Substrates efficiently converted by this yeast include the pentoses xylose and arabinose. Genetic engineering of C. lignohabitans with the isolated endogenous GAP promoter and GAP terminator was successful. GFP expression was used as a proof of functionality for the isolated transcription elements. Expression of lactate dehydrogenase and cis-aconitate decarboxylase resulted in stable and reproducible production of lactic acid and itaconic acid, respectively. The desired organic acids were accumulated converting pure sugars as well as lignocellulosic hydrolysates. C. lignohabitans proved therefore to be a promising reliable microbial host for production of organic acids from lignocellulosic material. PMID:25651876

  2. A Single-Use Paper-Shaped Microbial Fuel Cell for Rapid Aqueous Biosensing.

    PubMed

    Zuo, Kuichang; Liu, Han; Zhang, Qiaoying; Liang, Peng; Huang, Xia; Vecitis, Chad D

    2015-06-22

    The traditional chamber-based microbial fuel cell (MFC) often has the disadvantages of high ohmic resistance, large volume requirements, and delayed start-up. In this study, paper-shaped MFCs utilizing a porous carbon anode, a solid Ag2 O-coated carbon cathode, and a micrometer-thin porous polyvinylidene fluoride (PVDF) separator are investigated to address the classical MFC issues. The Ag2 O-coated cathode has a low overpotential of 0.06 V at a reducing current of 1 mA compared to a Pt-air cathode. Rapid inoculation by filtration results in an instantaneous power density of 92 mW m(-2) with an internal resistance of 162 Ω. Integrated current over the first 30 min of operation has a linear relation with microbial concentration. PMID:26013975

  3. Optimization of membrane stack configuration in enlarged microbial desalination cells for efficient water desalination

    NASA Astrophysics Data System (ADS)

    Chen, Xi; Sun, Haotian; Liang, Peng; Zhang, Xiaoyuan; Huang, Xia

    2016-08-01

    Microbial desalination cells are considered a low-energy-consumption, clean technology to simultaneously purify wastewater and desalinate saline water by utilizing the in situ energy source contained in wastewater. To enhance desalination performance and achieve an optimal membrane stack configuration, an enlarged stacked microbial desalination cell (SMDC) has been developed and tested with 6-14 desalination cells. The cross-membrane area of the enlarged SMDC is 100 cm2. The anode and cathode volumes are both 200 mL. To reduce internal resistance, the width of desalination cells is kept as <0.5 mm. The optimal configuration with 10 desalination cells achieves the highest total desalination rate (TDR) of 423 mg/h and the highest charge transfer efficiency (CTE) of 836% when treating the 20 g/L NaCl solution. During this process, the junction potential across membranes increases from 0 to 374 mV, and occupies up to 74% of the total potential loss inside the SMDC. This shows that the SMDC used in this work achieves the highest TDR and CTE among the reported studies, and the junction potential should be effectively controlled to achieve the desired desalination performance in future practical applications.

  4. Natural Killer-like B Cells Prime Innate Lymphocytes against Microbial Infection.

    PubMed

    Wang, Shuo; Xia, Pengyan; Chen, Yi; Huang, Guanling; Xiong, Zhen; Liu, Jing; Li, Chong; Ye, Buqing; Du, Ying; Fan, Zusen

    2016-07-19

    Natural killer (NK) cells and non-cytotoxic interferon-γ (IFN-γ)-producing group I innate lymphoid