Naoya Ohmura

Anaerobic respiration using Fe(3+), S(0), and H(2) in the chemolithoautotrophic bacterium Acidithiobacillus ferrooxidans

The chemolithoautotrophic bacterium Acidithiobacillus ferrooxidans has been known as an aerobe that respires on iron and sulfur. Here we show that the bacterium could chemolithoautotrophically grow not only on H(2)/O(2) under aerobic conditions but also on H(2)/Fe(3+), H(2)/S(0), or S(0)/Fe(3+) under anaerobic conditions. Anaerobic respiration using Fe(3+) or S(0) as an electron acceptor and H(2) or S(0) as an electron donor serves as a primary energy source of the bacterium. Anaerobic respiration based on reduction of Fe(3+) induced the bacterium to synthesize significant amounts of a c-type cytochrome that was purified as an acid-stable and soluble 28-kDa monomer. The purified cytochrome in the oxidized form was reduced in the presence of the crude extract, and the reduced cytochrome was reoxidized by Fe(3+). Respiration based on reduction of Fe(3+) coupled to oxidation of a c-type cytochrome may be involved in the primary mechanism of energy production in the bacterium on anaerobic iron respiration.

Isolation and determination of cultural characteristics of microalgae which functions under CO2 enriched atmosphere

A fresh-water microalgae, which functions under CO2 enriched atmosphere conditions, was isolated and its cultural characteristics were investigated. The HA-1 strain, identified as genus Chlorella, was newly isolated from a paddy field by an enrichment culture using reproduced stack gases from a thermal power plant with a concentration of CO2 and O2 of 15% and 2% respectively. It showed maximum growth at 10% CO2 enriched air flowing condition, and showed a good growth rate in a broad range of physically controllable conditions, including CO2 enriched air flow rate, temperature and pH value. The results indicated the feasibility of the HA-1 strain for mass cultivation using stack gases.

Bioelectrochemical system stabilizes methane fermentation from garbage slurry.

Methanogenic bioreactors, which are packed with supporting material, have attracted attention as an efficient means of degrading garbage. We aimed to increase bioreactor performance by using an electrochemical system to regulate the electrical potential on supporting material. At an organic loading rate of 26.9g dichromate chemical oxygen demand (CODcr)/L/day, reactors with a potential of -0.6 or -0.8V, generated by a cathodic electrochemical reaction, showed greater removal of CODcr and methanogenesis than reactors with a potential of 0.0 or -0.3V, generated by anodic reaction, or control reactors without electrochemical regulation. 16S rRNA gene analysis revealed that the same methanogens were present in all our reactors, but quantitative real-time polymerase chain reaction showed that higher prokaryotic and methanogenic copy numbers were present on cathodic electrodes than on anodic or control electrodes. These results indicate that cathodic electrochemical regulation can support methane fermentation from garbage.

Syntrophic degradation of proteinaceous materials by the thermophilic strains Coprothermobacter proteolyticus and Methanothermobacter thermautotrophicus

Protein is a major component of organic solid wastes, and therefore, it is necessary to further elucidate thermophilic protein degradation process. The effects of hydrogenotrophic methanogens on protein degradation were investigated using the proteolytic bacterial strain CT-1 that was isolated from a methanogenic thermophilic (55°C) packed-bed reactor degrading artificial garbage slurry. Strain CT-1 was closely related to Coprothermobacter proteolyticus, which is frequently found in methanogenic reactors degrading organic solid wastes. Strain CT-1 was cultivated in the absence or presence of Methanothermobacter thermautotrophicus by using 3 kinds of proteinaceous substrates. Degradation rates of casein, gelatin, and bovine serum albumin were higher in co-cultures than in monocultures. Strain CT-1 showed faster growth in co-cultures than in monocultures. M. thermautotrophicus comprised 5.5-6.0% of the total cells in co-culture. Increased production of ammonia and acetate was observed in co-cultures than in monocultures, suggesting that addition of M. thermautotrophicus increases the products of protein degradation. Hydrogen produced in the monocultures was converted to methane in co-cultures. These results suggest that thermophilic proteolytic bacteria find it favorable to syntrophically degrade protein in a methanogenic environment, and that it is important to retain hydrogen-scavenging methanogens within the reactor.

Isolation and identification of a novel strain of the genus Ochrobactrum with phenol-degrading activity

A bacterial strain AS1 belonging to the genus Ochrobactrum, was isolated from an enriched phenol-activated sludge in Egypt. This strain grew aerobically on phenol as the sole carbon source using the meta-cleavage pathway at high phenol-degrading rates compared with those in a previous report.

Effect of adding carbon fiber textiles to methanogenic bioreactors used to treat an artificial garbage slurry

To compare the performances and microbial populations of methanogenic reactors with and without carbon fiber textiles (CFT), we operated small-scale (200 ml) reactors using a slurry of artificial garbage. For both types of reactors, the organic loading rate (OLR) was stepwisely and rapidly increased in the same manner. Start-up period was shortened by adding CFT. Reactors with CFT showed greater efficiency for removal of suspended solid and volatile suspended solid than reactors without CFT at a long hydraulic retention time (HRT) between 8 and 13 days. The reactors with CFT maintained stable methane production at an OLR of 15.3 g dichromate chemical oxygen demand (CODcr)/l/day and DNAs from microorganisms were highly concentrated in adhering fractions on CFT. As shown by quantitative PCR analysis, the proportions of methanogenic archaea were conserved more than 25% in adhering fractions on CFT in reactors with CFT. By contrast, reactors without CFT showed accumulation of volatile fatty acid and deteriorated at an OLR of 2.4 gCODcr/l/day. Methanogenic proportions dropped to 17.1% in suspended fractions of reactors without CFT. Denaturing gradient gel electrophoresis (DGGE) analysis revealed that all archaeal DGGE bands in both types of reactors were related to methanogens, but more bands were observed in reactors with CFT. Thus the higher performance of reactors with CFT likely reflects the greater abundance of microorganisms and methanogenic diversity.

Isolation and Characterization of Phenol-catabolizing Bacteria from a Coking Plant

New phenol degrading bacteria with high biodegradation activity and high tolerance were isolated as Burkholderia cepacia PW3 and Pseudomonas aeruginosa AT2. Both isolates could grow aerobically on phenol as a sole carbon source even at 3 g/l. The whole-cell kinetic properties for phenol degradation by strains PW3 and AT2 showed a Vmax of 0.321 and 0.253 mg/l/min/(mg protein), respectively. The metabolic pathways for phenol biodegradation in both strains were assigned to the meta-cleavage activity of catechol 2,3-dioxygenase.

Trace-Level Mercury Ion (Hg2+) Analysis in Aqueous Sample Based on Solid-Phase Extraction Followed by Microfluidic Immunoassay

Mercury is considered the most important heavy-metal pollutant, because of the likelihood of bioaccumulation and toxicity. Monitoring widespread ionic mercury (Hg(2+)) contamination requires high-throughput and cost-effective methods to screen large numbers of environmental samples. In this study, we developed a simple and sensitive analysis for Hg(2+) in environmental aqueous samples by combining a microfluidic immunoassay and solid-phase extraction (SPE). Using a microfluidic platform, an ultrasensitive Hg(2+) immunoassay, which yields results within only 10 min and with a lower detection limit (LOD) of 0.13 μg/L, was developed. To allow application of the developed immunoassay to actual environmental aqueous samples, we developed an ion-exchange resin (IER)-based SPE for selective Hg(2+) extraction from an ion mixture. When using optimized SPE conditions, followed by the microfluidic immunoassay, the LOD of the assay was 0.83 μg/L, which satisfied the guideline values for drinking water suggested by the United States Environmental Protection Agency (USEPA) (2 μg/L; total mercury), and the World Health Organisation (WHO) (6 μg/L; inorganic mercury). Actual water samples, including tap water, mineral water, and river water, which had been spiked with trace levels of Hg(2+), were well-analyzed by SPE, followed by microfluidic Hg(2+) immunoassay, and the results agreed with those obtained from reduction vaporizing-atomic adsorption spectroscopy.

Composition of the sheath produced by the green alga Chlorella sorokiniana

Aims:  To investigate the chemical characterization of the mucilage sheath produced by Chlorella sorokiniana.

Acidianus manzaensis sp. nov., a Novel Thermoacidophilic Archaeon Growing Autotrophically by the Oxidation of H2 with the Reduction of Fe3+

A novel thermoacidophilic iron-reducing Archaeon, strain NA−1, was isolated from a hot fumarole in Manza, Japan. Strain NA-1 could grow autotrophically using H2 or S0 as an electron donor and Fe3+ as an electron acceptor, and also could grow heterotrophically using some organic compounds. Fe3+ and O2 served as electron acceptors for growth. However, S0, NO3−, NO2−, SO42−, Mn4+, fumarate, and Fe2O3 did not serve as electron acceptors. The ranges of growth temperature and pH were 60–90°C (optimum: 80°C) and pH 1.0–5.0 (optimum: pH 1.2–1.5), respectively. Cells were nearly regular cocci with an envelope comprised of the cytoplasmic membrane and a single outer S-layer. The crenarchaeal-specific quinone (cardariellaquinone) was detected, and the genomic DNA G + C content was 29.9 mol%. From 16S rDNA analysis, it was determined that strain NA-1 is closely related to Acidianus ambivalens (93.1%) and Acidianus infernus (93.0%). However, differences revealed by phylogenetic and phenotypic analyses clearly show that strain NA-1 represents a new species, Acidianus manzaensis, sp. nov., making it the first identified thermoacidophilic iron-reducing microorganism (strain NA-1T = NBRC 100595 = ATCC BAA 1057).