Characterization of Biogenic PbS Quantum Dots.

Heavy metals in a polluted environment are toxic to life. However, some microorganisms can remove or immobilize heavy metals through biomineralization. These bacteria also form minerals with compositions similar to those of semiconductors. Here, this bioprocess was used to fabricate semiconductors with low energy consumption and cost. Bacteria that form lead sulfide (PbS) nanoparticles were screened, and the crystallinity and semiconductor properties of the resulting nanoparticles were characterized. Bacterial consortia that formed PbS nanoparticles were obtained. Extracellular particle size ranged from 3.9 to 5.5 nm, and lattice fringes were observed. The lattice fringes and electron diffraction spectra corresponded to crystalline PbS. The X-ray diffraction (XRD) patterns of bacterial PbS exhibited clear diffraction peaks. The experimental and theoretical data of the diffraction angles on each crystal plane of polycrystalline PbS were in good agreement. Synchrotron XRD measurements showed no crystalline impurity-derived peaks. Thus, bacterial biomineralization can form ultrafine crystalline PbS nanoparticles. Optical absorption and current-voltage measurements of PbS were obtained to characterize the semiconductor properties; the results showed semiconductor quantum dot behavior. Moreover, the current increased under light irradiation when PbS nanoparticles were used. These results suggest that biogenic PbS has band gaps and exhibits the general fundamental characteristics of a semiconductor.

Reaction Rate of Hydrothermal Ammonia Production from Chicken Manure.

Ammonia is an important fertilizer feedstock and an expected next-generation hydrogen carrier. Thus, it is necessary to ensure effective production of ammonia from the waste biomass. In this regard, chicken manure was treated in an autoclave under hydrothermal reaction conditions, and the ammonia release rate was determined in the temperature range of 250-400 °C for holding times ranging from 2 to 120 min. A reaction network for ammonia production was proposed, and the reaction rate constants were determined. A nitrogen yield as high as 0.8 was obtained, corresponding to a hydrogen potential of 88.1 billion m3/year from chicken manure. Consequently, chicken manure was identified as a potentially favorable feedstock for ammonium production.

Reutilization of Algal Supercritical Water Gasification Waste for Microalgae Chlorella vulgaris Cultivation.

Effluents obtained through a supercritical water gasification (SCWG) process at 400 and 600 °C were mixed with Bristol Medium to cultivate Chlorella vulgaris. Improvement of growth rate was observed only for the medium with the effluent at 600 °C. Low non-purgeable organic carbon implied that the inhibiting material was decomposed due to the high temperature of 600 °C. Thus, SCWG effluents might be more suitable for algae cultivation than hydrothermal liquefaction effluents. Phosphorus accumulation in C. vulgaris was improved in the SCWG mixed medium, irrespective of the treatment temperature. The media with SCWG effluents showed 2.5 times higher phosphorus accumulation in the algae, indicating the possibility of using a combination of C. vulgaris and SCWG for nutrient recycling processes.

Feasible conditions for Japanese woody biomass utilization.

There is an abundance of woody biomass in Japan. However, its economic feasibility is limited. There have been several discussions on whether generation of heat or electrical power is more suitable for woody biomass. In this study, we clarified the feasibility of generating heat and electricity from the viewpoint of biomass unit price and biomass productivity based on actual operation data for the first time. It was determined that heat production is feasible for a small-scale biomass energy use of 2,660 t/year or more. Electrical power generation is feasible only for a large-scale biomass energy use of 13,100 t/year or more, but it allows use of woody biomass with a higher unit price.

Supercritical Water Gasification of Guaiacol with Acetic Acid as a Radical Scavenger: Interaction Effect on Char Formation and Gas Composition.

Supercritical water gasification (SCWG) of mixtures of guaiacol and acetic acid was carried out in a continuous reactor at 600 °C and 25 MPa with a residence time of 94 s. Different concentrations of acetic acid were employed to investigate the effect of acetic acid on product yield and gas composition. The interaction between guaiacol and acetic acid during SCWG was discussed. Acetic acid, as a radical scavenger, was found to inhibit radical polymerization, resulting in the suppression of char formation.

Response of Palladium and Carbon Nanotube Composite Films to Hydrogen Gas and Behavior of Conductive Carriers.

To develop a high-performance hydrogen gas sensor, we fabricated a composite film made of carbon nanotubes (CNTs) and palladium nanoparticles. Carbon nanotubes were spin-coated onto a glass substrate, and subsequently, palladium nanoparticles were sputtered onto this film. The response to hydrogen gas was measured during two seasons (summer and winter) using a vacuum chamber by introducing a hydrogen/argon gas mixture. There was a clear difference in the sensor response despite the temperature difference between summer and winter. In addition, since a clean chamber was used, fewer water molecules acted as a dopant, and the behavior of the CNT changed from p-type to n-type because of the dissociative adsorption of hydrogen. This phenomenon was confirmed as the Seebeck effect. Finally, the work functions of Pd, PdHx, and CNT were calculated by first-principle calculations. As predicted by previous studies, a decrease in work function due to hydrogen adsorption was confirmed; however, the electron transfer to CNT was not appropriate from the perspective of charge neutrality and was found to be localized at the Pd/CNT interface. It seems that the Seebeck effect causes the concentration of conductive carriers to change.

Flocculation of Chlorella vulgaris by shell waste-derived bioflocculants for biodiesel production: Process optimization, characterization and kinetic studies.

Flocculants are foreign particles that aggregate suspended microalgae cells and due to cost factor and toxicity, harvesting of microalgae biomass has shifted towards the use of bioflocculants. In this study, mild acid-extracted bioflocculants from waste chicken's eggshell and clam shell were used to harvest Chlorella vulgaris that was cultivated using chicken compost as nutrient source. It was found that a maximum of 99% flocculation efficiency can be attained at pH medium of 9.8 using 60 mg/L of hydrochloric acid-extracted chicken's eggshell bioflocculant at 50 °C of reaction temperature. On the other hand, 80 mg/L of hydrochloric acid-extracted clam shell bioflocculant was sufficient to recover C. vulgaris biomass at pH 9.8 and optimum temperature of 40 °C. The bioflocculants and bioflocs were characterized using microscopic, zeta potential, XRD, AAS and FT-IR analysis. The result revealed that calcium ions in the bioflocculants are the main contributor towards the flocculation of C. vulgaris, employing charge neutralization and sweeping as possible flocculation mechanisms. The kinetic parameters were best fitted pseudo-second order which resulted in R2 of 0.99 under optimal flocculation temperature. The results herein, disclosed the applicability of shell waste-derived bioflocculants for up-scaled microalgae harvesting for biodiesel production.

Complete genome sequence of Nitratireductor sp. strain OM-1: A lipid-producing bacterium with potential use in wastewater treatment.

Reducing CO2 emissions is necessary to alleviate rising global temperature. Renewable sources of energy are becoming an increasingly important substitute for fossil fuels. An important step in this direction is the isolation of novel, technologically relevant microorganisms. Nitratireductor sp. strain OM-1 can convert volatile short-chain fatty acids in wastewater into 2-butenoic acid and its ester and can accumulate intracellularly esterified compounds up to 50% of its dried cell weight under nitrogen-depleted conditions. It is believed that a novel fatty acid biosynthesis pathway including an esterifying enzyme is encoded in its genome. In this study, we report the whole-genome sequence (4.8 Mb) of OM-1, which comprises a chromosome (3,977,827 bp) and a megaplasmid (857,937 bp). This sequence information provides insight into the genome organization and biochemical pathways of OM-1. In addition, we identified lipid biosynthesis pathways in OM-1, paving the way to a better understanding of its biochemical characterization.

RNase H-assisted RNA-primed rolling circle amplification for targeted RNA sequence detection.

RNA-primed rolling circle amplification (RPRCA) is a useful laboratory method for RNA detection; however, the detection of RNA is limited by the lack of information on 3'-terminal sequences. We uncovered that conventional RPRCA using pre-circularized probes could potentially detect the internal sequence of target RNA molecules in combination with RNase H. However, the specificity for mRNA detection was low, presumably due to non-specific hybridization of non-target RNA with the circular probe. To overcome this technical problem, we developed a method for detecting a sequence of interest in target RNA molecules via RNase H-assisted RPRCA using padlocked probes. When padlock probes are hybridized to the target RNA molecule, they are converted to the circular form by SplintR ligase. Subsequently, RNase H creates nick sites only in the hybridized RNA sequence, and single-stranded DNA is finally synthesized from the nick site by phi29 DNA polymerase. This method could specifically detect at least 10 fmol of the target RNA molecule without reverse transcription. Moreover, this method detected GFP mRNA present in 10 ng of total RNA isolated from Escherichia coli without background DNA amplification. Therefore, this method can potentially detect almost all types of RNA molecules without reverse transcription and reveal full-length sequence information.

Isolation of High Carotenoid-producing Aurantiochytrium sp. Mutants and Improvement of Astaxanthin Productivity Using Metabolic Information.

The marine eukaryotic microheterotroph thraustochytrid genus Aurantiochytrium is a known producer of polyunsaturated fatty acids, carotenoids, and squalene. We previously constructed a lipid fermentation system for Aurantiochytrium sp. strains using underutilized biomass, such as canned syrup and brown macroalgae. To improve the productivity, in this study, Aurantiochytrium sp. RH-7A and RH-7A-7 that produced high levels of carotenoids, such as astaxanthin and canthaxanthin, were isolated through chemical mutagenesis. Moreover, metabolomic analysis of the strain RH-7A revealed that oxidative stress impacts carotenoid accumulation. Accordingly, the addition of ferrous ion (Fe2+), as an oxidative stress compound, to the culture medium significantly enhanced the production of astaxanthin by the mutants. These approaches improved the productivity of astaxanthin up to 9.5 mg/L/day at the flask scale using not only glucose but also fructose which is the main carbon source in fermentation systems with syrup and brown algae as the raw materials.

Comparative study of hydrothermal pretreatment for rice straw and its corresponding mixture of cellulose, xylan, and lignin.

We herein investigated the effect of hemicellulose and lignin on the hydrothermal pretreatment of lignocellulosic biomass for glucose production from cellulose in terms of structural cross-linking between cellulose and the above components. A comparison was made between the hydrothermal pretreatment of biomass and a mixture containing the individual model compounds in the same composition ratio. Thus, rice straw biomass and the mixture containing cellulose, xylan, and lignin were treated in an autoclave reactor at temperatures between 150 and 250 °C for 30 min. The obtained products were then subjected to enzymatic hydrolysis. Interestingly, different results were obtained for the two samples, as the presence of cross-linking between cellulose and lignin in the original biomass sample affected the efficiency of hydrothermal pretreatment. A model was therefore proposed to account for the obtained result in accordance with previous knowledge regarding the behavior of these compounds under hydrothermal conditions.

Supercritical water gasification of sewage sludge in continuous reactor.

In this study, a process for the continuous recovery of phosphorus and generation of gas from sewage sludge is investigated for the first time using supercritical water gasification (SCWG). A continuous reactor was employed and experiments were conducted by varying the temperature (500-600 °C) and residence time (5-60 s) while fixing the pressure at 25 MPa. The behavior of phosphorus during the SCWG process was studied. The effect of the temperature and time on the composition of the product gas was also investigated. A model of the reaction kinetics for the SCWG of sewage sludge was developed. The organic phosphorus (OP) was rapidly converted into inorganic phosphorus (IP) within a short residence time of 10 s. The gaseous products were mainly composed of H2, CO2, and CH4. The reaction followed first order kinetics, and the model was found to fit the experimental data well.

Efficient conversion of mannitol derived from brown seaweed to fructose for fermentation with a thraustochytrid.

Macroalgae are a promising biomass feedstock for energy and valuable chemicals. Mannitol and alginate are the major carbohydrates found in the microalga Laminaria japonica (Konbu). To convert mannitol to fructose for its utilization as a carbon source in mannitol non-assimilating bacteria, a psychrophile-based simple biocatalyst (PSCat) was constructed using a psychrophile as a host by expressing mesophilic enzymes, including mannitol 2-dehydrogenase for mannitol oxidation, and NADH oxidase and alkyl hydroxyperoxide reductase for NAD+ regeneration. PSCat was treated at 40 °C to inactivate the psychrophilic enzymes responsible for byproduct formation and to increase the membrane permeability of the substrate. PSCat efficiently converted mannitol to fructose with high conversion yield without additional input of NAD+. Konbu extract containing mannitol was converted to fructose with hydroperoxide scavenging, inhibiting the mannitol dehydrogenase activity. Auranthiochytrium sp. could grow well in the presence of fructose converted by PSCat. Thus, PSCat is a potential carbohydrate converter for mannitol non-assimilating microorganism.

Improved methanization and microbial diversity during batch mode cultivation with repetition of substrate addition using defined organic matter and marine sediment inoculum at seawater salinity.

The activation of microbes, which are needed to initiate continuous methane production, can be accomplished by fed-batch methanization. In the present study, marine sediment inoculum was activated by batch mode methanization with repetition of substrate addition using defined organic matter from sugar, protein, or fat at seawater salinity to investigate the potential for application of the activation method to various types of saline waste and microbial community compositions. All substrates had methane potentials close to the theoretical value except for bovine serum albumin (BSA) whose methane potential was lower, but the maximum methane potential reached the value during repeated methanization. Beta diversity analysis revealed that substrate (especially BSA)-fed and non-fed cultures had distinct microbial community compositions. Bacterial members depended on substrate. Thus, marine sediment inocula activated via the methanization method can be used to effectively treat various types of saline waste.

Continuous production of biodiesel under supercritical methyl acetate conditions: Experimental investigation and kinetic model.

In this study, biodiesel production by using supercritical methyl acetate in a continuous flow reactor was investigated for the first time. The aim of this study was to elucidate the reaction kinetics of biodiesel production by using supercritical methyl. Experiments were conducted at various reaction temperatures (300-400°C), residence times (5-30min), oil-to-methyl acetate molar ratio of 1:40, and a fixed pressure of 20MPa. Reaction kinetics of biodiesel production with supercritical methyl acetate was determined. Finally, biodiesel yield obtained from this method was compared to that obtained with supercritical methanol, ethanol, and MTBE (methyl tertiary-butyl ether). The results showed that biodiesel yield with supercritical methyl acetate increased with temperature and time. The developed kinetic model was found to fit the experimental data well. The reactivity of supercritical methyl acetate was the lowest, followed by that of supercritical MTBE, ethanol, and methanol, under the same conditions.

Bacterial community structure and predicted alginate metabolic pathway in an alginate-degrading bacterial consortium.

Methane fermentation is one of the effective approaches for utilization of brown algae; however, this process is limited by the microbial capability to degrade alginate, a main polysaccharide found in these algae. Despite its potential, little is known about anaerobic microbial degradation of alginate. Here we constructed a bacterial consortium able to anaerobically degrade alginate. Taxonomic classification of 16S rRNA gene, based on high-throughput sequencing data, revealed that this consortium included two dominant strains, designated HUA-1 and HUA-2; these strains were related to Clostridiaceae bacterium SK082 (99%) and Dysgonomonas capnocytophagoides (95%), respectively. Alginate lyase activity and metagenomic analyses, based on high-throughput sequencing data, revealed that this bacterial consortium possessed putative genes related to a predicted alginate metabolic pathway. However, HUA-1 and 2 did not grow on agar medium with alginate by using roll-tube method, suggesting the existence of bacterial interactions like symbiosis for anaerobic alginate degradation.

Isolation and characterization of bacterium producing lipid from short-chain fatty acids.

Anaerobic fermentation generates propionic acid, which inhibits microbial growth and accumulates in wastewater containing increased amounts of organic matter. We therefore isolated a propionic acid-assimilating bacterium that could produce triacylglycerol, for use in wastewater treatment. Nitratireductor sp. strain OM-1 can proliferate in medium containing propionic, acetic, butyric, and valeric acids as well as glycerol, and produces triacylglycerol when both propionic and acetic acids or glycerol are present. In composite model wastewater containing acetic acid, propionic acid and glycerol, this strain shows an even higher conversion rate, suggesting that it is suitable for wastewater treatment. Further, nitrogen depletion in medium containing an acetic-propionic acid mixture resulted in the production of the light oil 2-butenoic acid 1-methylethyl ester, but not triacylglycerol. Collectively, our data indicate that strain OM-1 has the potential to reduce accumulation of activated sludge in wastewater treatment and may contribute to the production of biodiesel.

Semi-continuous methane production from undiluted brown algae using a halophilic marine microbial community.

Acclimated marine sediment-derived culture was used for semi-continuous methane production from materials equivalent to raw brown algae, without dilution of salinity and without nutrient supply, under 3 consecutive conditions of varying organic loading rates (OLRs) and hydraulic retention time (HRT). Methane production was stable at 2.0gVS/kg/day (39-day HRT); however, it became unstable at 2.9gVS/kg/day (28-day HRT) due to acetate and propionate accumulation. OLR subsequently decreased to 1.7gVS/kg/day (46-day HRT), stabilizing methane production beyond steady state. Methane yield was above 300mL/g VS at all OLRs. These results indicated that the acclimated marine sediment culture was able to produce methane semi-continuously from raw brown algae without dilution and nutrient supply under steady state. Microbial community analysis suggested that hydrogenotrophic methanogens predominated among archaea during unstable methane production, implying a partial shift of the methanogenic pathway from acetoclastic methanogenesis to acetate oxidation.

Characterization of a halotolerant acetoclastic methanogen highly enriched from marine sediment and its application in removal of acetate.

A marine sediment collected from Hiroshima Bay was cultured in artificial seawater, containing 0.51 M NaCl and 60 mM acetate and was found to exhibit active methane production at 37°C. Following four successive serial dilutions of cultures in medium containing 0.51 M NaCl, 60 mM acetate, and antibiotics, the well-acclimated methanogen was found to exhibit growth over a range of NaCl concentration (between 0 M and 2.06 M). The specific growth rates of the highly enriched methanogen, termed strain HA, in the absence of NaCl and in the presence of 1.54 M NaCl were estimated to be 0.037 h(-1) and 0.027 h(-1), respectively. The pH and temperature for optimum growth were determined to be 7.0-8.8 and 37°C, respectively. Although cells that had morphology similar to Methanosaeta sp. became dominant in the culture, methane production was still detected in the medium containing 0.51 M NaCl and other substrates such as methanol, formate, and methylamine, indicating contamination with other methanogens. The phylogenetic tree based on 16S rRNA gene sequences revealed that the strain HA was closely related to Methanosaeta harundinacea 6Ac and 8Ac(T), with sequence similarity of 98% and 97%, respectively. The continuous removal of acetate with upflow anaerobic filter reactor for industrial use of strain HA determined a methane production rate of 70 mM/d under condition of 0.51 M NaCl and successful methane production even under 1.54 M NaCl.

Effect of salinity on methanogenic propionate degradation by acclimated marine sediment-derived culture.

Degradation of propionate under high salinity is needed for biomethane production from salt-containing feedstocks. In this study, marine sediment-derived culture was evaluated to determine the effect of salinity on methanogenic propionate degradation. Microbes in marine sediments were subjected to fed-batch cultivation on propionate for developing acclimatized cultures. The rate of propionate degradation increased eightfold during 10 rounds of cultivation. Microbial community composition was determined through pyrosequencing of 16S rRNA gene amplicons after 10 rounds of cultivation. Taxa analysis was conducted for the reads obtained by pyrosequencing. Known propionate degraders were undetectable in the acclimated culture. Comparison of bacterial taxa in the original sediment with those in the acclimated culture revealed that the populations of four bacterial taxa were significantly increased during acclimation. Methanolobus was the predominant archaea genus in the acclimated culture. The propionate degradation rate of the acclimated culture was not affected by salinity of up to equivalent of 1.9 % NaCl. The rate decreased at higher salinity levels and was more than 50 % of the maximum rate even at equivalent of 4.3 % NaCl.