Hayato Tokumoto

Efficient, high-speed methane fermentation for sewage sludge using subcritical water hydrolysis as pretreatment.

A novel biomass-energy process for the production of methane from sewage sludge using a subcritical water (sub-CW) hydrolysis reaction as pretreatment is proposed. The main substances of sewage sludge hydrolyzed by sub-CW at 513 K for 10 min were acetic acid, formic acid, pyroglutamic acid, alanine, and glycine. Fermentation experiments were conducted in an anaerobic-sludge reactor for two different samples: real sewage sludge and a model solution containing components typically produced by the sub-CW pretreatment of sewage sludge. In the experiment for the sub-CW pretreatment of sewage sludge, methane generation was twice that for non-pretreatment after 3 days of incubation. In the model experiment, the methane conversion was about 40% with the application of mixture of organic acids and amino acids after 5 days of incubation. Furthermore, the methane conversion was about 60% for 2 days when only organic acids, such as acetic acid and formic acid, were applied. Because acetic acid is the key intermediate and main precursor of the methanogenesis step, fermentation experiments were conducted in an anaerobic-sludge reactor with high concentrations of acetic acid (0.01-0.1M). Nearly 100% of acetic acid was converted to methane and carbon dioxide in 1-3 days.

Exposure of the yeast Saccharomyces cerevisiae to functionalized polystyrene latex nanoparticles: influence of surface charge on toxicity.

Novel nanoparticles with unique physicochemical characteristics are being developed with increasing frequency, leading to higher probability of nanoparticle release and environmental accumulation. Therefore, it is important to assess the potential environmental and biological adverse effects of nanoparticles. In this study, we investigated the toxicity and behavior of surface-functionalized nanoparticles toward yeast (Saccharomyces cerevisiae). The colony count method and confocal microscopy were used to examine the cytotoxicity of manufactured polystyrene latex (PSL) nanoparticles with various functional groups (amine, carboxyl, sulfate, and nonmodified). S. cerevisiae were exposed to PSL nanoparticles (40 mg/L) dispersed in 5-154 mM NaCl solutions for 1 h. Negatively charged nanoparticles had little or no toxic effect. Interestingly, nanoparticles with positively charged amine groups (p-Amine) were not toxic in 154 mM NaCl, but highly toxic in 5 mM NaCl. Confocal microscopy indicated that in 154 mM NaCl, the p-Amine nanoparticles were internalized by endocytosis, whereas in 5 mM NaCl they covered the dead cell surfaces. This demonstrates that nanoparticle-induced cell death might to be related to their adhesion to cells rather than their internalization. Together, these findings identify important factors in determining nanoparticle toxicity that might affect their impact on the environment and human health.

Novel anaerobic digestion induced by bacterial components for value-added byproducts from high-loading glycerol.

A novel bioprocessing system was developed and tested that involved anaerobic fermentation to degrade high-loading glycerol by a fermentation promoter, and which could be used for the production of important resources. In the absence of a promoter, there was no anaerobic digestion when glycerol (4.0-6.0%, v/v) was added to the reactor. By contrast, glycerol was readily decomposed when sewage sludge, acting as a fermentation promoter, was added to the anaerobic reactor. Fermentation resulted in the generation of hydrogen, 1,3-propanediol (1,3-PDO) and various organic acids. In 7 days, glycerol decomposition reached 88%; hydrogen production was 3.1mg/kg-glycerol (0.0004 g/day/L), and 1,3-PDO yield reached 0.35 kg/kg-glycerol (0.05 g/day/L). Further experiments confirmed that the bacteria Escherichia coli and particularly Schizosaccharomyces pombe (found within sewage sludge) and especially glucose (found within bacterial components (i.e., cell walls)) acted as efficient promoters of fermentation.

Effect of the surface characteristics of Methanosarcina barkeri on immobilization to support materials

Abstract To realize a highly efficient anaerobic treatment, it is necessary to immobilize high concentrations of methanogens within a fermenter. In this study, we experimentally examine the effect of the surface characteristics of the acetate-utilizing methanogen Methanosarcina barkeri (JCM 10043) on immobilization to support materials. To this aim, we measured the electrostatic and hydrophobic properties of M. barkeri . The electrophoretic mobility of M. barkeri decreased with increasing ionic strength of the cell suspension and was fitted by the Ohshima equation using two parameters: spatial charge density in the polyelectrolyte region ( ZeN = –1.15 × 10 6 C/m 3 ) and the softness parameter (1/λ = 3.35 × 10 −9 m). M. barkeri showed an affinity to n -hexadecane, with adhesion of more than 60%. M. barkeri showed hydrophobicity relative to Escherichia coli . We also carried out a microbial adhesion test to support materials. M. barkeri showed better adhesion to the anion-exchange resin than to the hydrophobic resin. The microbial cells adhered uniformly to the anion-exchange resin, while coagulated cells adhered non-uniformly to the hydrophobic resin. M. barkeri showed poor adhesion to the cation-exchange resin. The anion-exchange resin is most effective in immobilizing M. barkeri within the fermenter.

Measurement of microbial adhesive forces with a parallel plate flow chamber.

HYPOTHESIS It was predicted that the colloidal behaviors of archaea and bacteria with disparate surface structure were different. In this study, the effects of the physicochemical properties of microbial cell surfaces on colloidal behavior were analyzed with Derjaguin-Landau-Verwey-Overbeek (DLVO) theory, thermodynamics, and powder technology. EXPERIMENTS Cell attachment and detachment from model substrates were directly observed using a parallel plate flow chamber. Gram-negative Escherichia coli and archaeal Methanosarcina barkeri were used as model microbial cells, and positively and negatively charged glass slides were used as model substrates. FINDINGS Microbial adhesion on both substrates agreed well with predictions calculated from DLVO theory, using experimental parameters. The total number of cells detached from the substrates as a function of flow rate was fit with the Weibull distribution function. In addition, the drag force required for detachment, which was estimated from the hydrodynamic forces, had a wide distribution; however, the forces became smaller with increasing ionic strength because of reduced electrostatic interactions between the cells and the substrate. M. barkeri could not be detached from positively charged substrates because it would entail a negative change in the interfacial energy of interaction. Thus adhesion was thermodynamically favored in this case.

Value-adding conversion and volume reduction of sewage sludge by anaerobic co-digestion with crude glycerol

In this study, conversion of sewage sludge to biogas by anaerobic co-digestion with crude glycerol was examined. When 0.126g/L crude glycerol was added to the reactor, only methane was produced. Upon addition of 5.04g/L crude glycerol, hydrogen production occurred, and a significant amount of 1,3-propanediol (1,3-PDO) was generated in the liquid phase. On day 6, the dry weight was largely composed of organic acids (48%) and 1,3-PDO (17%), which are water-soluble. Degradation of 1,3-PDO was very slow, which is advantageous for recovery. Crude glycerol, which contains alkaline substances, promoted organic matter degradation by microorganisms, which possibly affected biogas and 1,3-PDO production. Addition of 0.630-2.52g/L glycerol initially led to hydrogen production, followed by methane production a few days later, which stabilized within 1week. In conclusion, adjustment of the crude glycerol concentration allows controllable conversion to value-added products for co-digestion.

Estimation of the adhesive force distribution for the flagellar adhesion of Escherichia coli on a glass surface

The effects of the presence or absence of microbial flagella and the microbial motility on the colloidal behaviors of microbial cells were quantitatively evaluated. The microbial cell attachment and detachment processes on a glass surface were observed directly using a parallel-plate flow chamber. Wild-type, flagellar paralyzed, and nonflagellated Escherichia coli strains were used as model microbial cells. In the cell attachment tests, the microbial adhesion rate in a 160mM NaCl solution was approximately 10 times higher than that in a 10mM solution, for all E. coli strains. The colloidal behavior of the microbial cells agreed well with the predictions of the DLVO theory. In addition, the microbial flagella and motility did not significantly affect the cell attachment, regardless of the existence of a potential barrier between the cell and the glass substratum. In the cell detachment tests, the cumulative number of microbial cells detached from the glass substratum with increasing flow rate was fit well with the Weibull distribution function. The list of strains arranged in order of increasing median drag force required to remove them was nonflagellated strain, flagellar paralyzed strain, and wild-type strain. These results indicated that the flagella and the flagellar motility inhibited the cell detachment from the glass substratum. Furthermore, a large external force would likely be required to inhibit the microbial adhesion in the early stage of the biofilm formation.

Changes in dominant fermentation type during anaerobic digestion of high-loading glycerol with slight glucose content

High-loading glycerol containing slight amounts of five different monosaccharides was inoculated with seed sludge obtained from a methane fermentation reactor. The use of different monosaccharides as fermentation promoters resulted in changes in fermentation types; in particular, glucose induced the formation of 1,3-propanediol. After 9 days incubation with glucose, glycerol levels had fallen by 81%, while molar yields of organic acids and 1,3-propanediol (per mole of glycerol degraded) were 0.22 and 0.39, respectively. Other monosaccharides enhanced methane production after 14 days of incubation in the following order: galactose, galacturonic acid, mannose and arabinose. Hydrogen was generated (together with a negligible amount of methane) only in the presence of glucose. When glucose was introduced to a methane-producing reactor (promoted by galacturonic acid), hydrogen production began 5 days later and displaced the methane production after 12 days. These results suggest that glucose catalyzes glycerol degradation, resulting in the production of hydrogen.

Bacterial Toxicity of Functionalized Polystyrene Latex Nanoparticles Toward Escherichia coli

Nanotechnology has the potential to produce a variety of new materials in the coming years, as a result of the design of novel nanoparticles with new physicochemical characteristics. However, their potential to adversely affect the environment and human health must be addressed. The toxicity of polystyrene latex (PSL) nanoparticles with various functional groups toward Escherichia coli KP7600 strain was investigated using the colony count method, and confocal microscopy observations. It was found that the positively charged PSL nanoparticles led to the death of the bacterial cells. Confocal observations of the bacterial cells after 1 h of exposure to the amine-modified, positively-charged PSL nanoparticles in an aqueous NaCl solution showed that the surfaces of the dead cells were almost entirely covered with the nanoparticles. No uptake of the nanoparticles into the bacterial cells was observed, regardless of the cell viability. It is likely that the adhesion of the positively charged nanoparticles onto the surface of the bacterial cells (due to the electrostatic attractive force) caused a decrease in the fluidity of the cell membrane, and the inhibition of metabolism through the cell membrane led to the death of the bacterial cells.

Synthesis of Hollow Titania Photocatalytic Particles Using Yeast as Templates

Hollow inorganic particles have attracted considerable interest for a variety of applications. In this work, yeast powder was used as a bio-template to fabricate yeast/titania core-shell spheres. A titania precursor was deposited via the hydrolysis of tetraisopropyl titanate. Triethanolamine was employed to control the reaction rate of the sol-gel process. The hollow titania particles with a diameter of approximately 4 m and a thickness of approximately 54 nm were successfully obtained after calcination of the core-shell spheres at 700 °C. The crystal structure of the particles was anatase. The specific surface area and the average pore diameter were 21.4 m2/g and 1.6 nm, respectively. The photocatalytic activity of the hollow particles was higher compared with that of solid particles.