Fumihisa Kobayashi

Phylogenetic analysis of atmospheric halotolerant bacterial communities at high altitude in an Asian dust (KOSA) arrival region, Suzu City.

The microbial communities transported by Asian desert dust (KOSA) events have attracted much attention as bioaerosols because the transported microorganisms are thought to influence the downwind ecosystems in Korea and Japan. However, the atmospheric microbial community has not been investigated at high altitude in the KOSA arrival area. In this study, to estimate the viability and diversity of atmospheric halotolerant bacteria, which are expected to resist to various environmental stresses as well as high salinities, bioaerosol samples were collected at 10 and 600 m above the ground within the KOSA arrival area, Suzu City, Japan, during KOSA events. During the sampling period, the particle numbers at 600 m were higher than those at 10 m, suggesting that large particles of aerosol fall from the high altitude of 600 m to the ground surface. The microorganisms in bioaerosol samples grew in media containing up to 15% NaCl concentrations demonstrating the viability of the halotolerant bacteria in bioaerosol samples. The PCR-DGGE analysis using 16S rDNA revealed that the bacterial species in NaCl-amended cultures were similar to the bacteria detected from the genomic DNA directly extracted from the bioaerosol samples. The 16S rDNA sequences of bacterial communities in bioaerosol samples were classified into 4 phylotypes belonging to the Bacilluscereus or Bacillussubtilis group. The bioaerosol samples collected at 600 m included 2 phylotypes belonging to B. subtilis, and one phylotype among all 4 phylotypes was identical between the samples at 10 and 600 m. In the atmosphere at 600 m, the halotolerant bacterial community was expected to remain viable, and the species composition was expected to include a few species of the genus Bacillus. During this investigation period, these atmospheric bacteria may have been vertically transported to the ground surface, where the long-range KOSA particle transport from China is frequently observed.

Alcohol fermentation of starch by a genetic recombinant yeast having glucoamylase activity

Alcohol fermentation of starch was investigated using a direct starch fermenting yeast, Saccharomyces cerevisiae SR93, constructed by integrating a glucoamylase-producing gene (STA1) into the chromosome of Saccharomyces cerevisiae SH1089. The glucoamylase was constitutively produced by the recombinant yeast. The ethanol concentration produced by the recombinant yeast was 14.3 g/L which was about 1.5-fold higher than by the conventional mixed culture using an amylolytic microorganism and a fermenting microorganism. About 60% of the starch was converted into ethanol by the recombinant yeast, and the ethanol yield reached its maximum value of 0.48 at the initial starch concentration of 50 g/L. The fed-batch culture, which maintains the starch concentration in the range of 30 to 50 g/L, was used to produce a large amount of ethanol from starch. The amount of ethanol produced in the fed-batch culture increased about 20% compared to the batch culture. (c) 1997 John Wiley & Sons, Inc.

Methane production from steam-exploded bamboo.

To convert unutilized plant biomass into a useful energy source, methane production from bamboo was investigated using a steam explosion pretreatment. Methane could not be produced from raw bamboo but methane production was enhanced by steam explosion. The maximum amount of methane produced, i.e., about 215 ml, was obtained from 1 g of exploded bamboo at a steam pressure of 3.53 MPa and a steaming time of 5 min. A negative correlation between the amount of methane produced and the amount of Klason lignin was observed in the methane fermentation of steam-exploded bamboo.

Variations in the structure of airborne bacterial communities in a downwind area during an Asian dust (Kosa) event

Asian dust (Kosa) events transport airborne microorganisms that significantly impact biological ecosystems, human health, and ice-cloud formation in downwind areas. However, the composition and population dynamics of airborne bacteria have rarely been investigated in downwind areas during Kosa events. In this study, air samplings were sequentially performed at the top of a 10-m high building within the Kosa event arrival area (Kanazawa City, Japan) from May 1 to May 7, 2011, during a Kosa event. The particle concentrations of bacterial cells and mineral particles were ten-fold higher during the Kosa event than on non-Kosa event days. A 16S ribosomal DNA clone library prepared from the air samples primarily contained sequences from three phyla: Cyanobacteria, Firmicutes, and Alphaproteobacteria. The clones from Cyanobacteria were mainly from a marine type of Synechococcus species that was dominant during the first phase of the Kosa event and was continuously detected throughout the Kosa event. The clones from Alphaproteobacteria were mainly detected at the initial and final periods of the Kosa event, and phylogenetic analysis showed that their sequences clustered with those from a marine bacterial clade (the SAR clade) and Sphingomonas spp. During the middle of the Kosa event, the Firmicutes species Bacillus subtilis and Bacillus pumilus were predominant; these species are known to be predominant in the atmosphere above the Chinese desert, which is the source of the dust during Kosa events. The clones obtained after the Kosa event had finished were mainly from Bacillus megaterium, which is thought to originate from local terrestrial areas. Our results suggest that airborne bacterial communities at the ground level in areas affected by Kosa events change their species compositions during a Kosa event toward those containing terrestrial and pelagic bacteria transported from the Sea of Japan and the continental area of China by the Kosa event.

Saccharification and alcohol fermentation in starch solution of steam-exploded potato.

Steam explosion pretreatment of potato for the efficient production of alcohol was experimentally studied. The amount of water-soluble starch increased with the increase of steam pressure, but the amounts of methanol-soluble material and Klason lignin remained insignificant, regardless of steam pressure. The potatoes exploded at high pressure were hydrolyzed into a low molecular liquid starch, and then easily converted into ethanol by simultaneous saccharification and fermentation using mixed microorganisms: an amylolytic microorganism,Aspergillus awamori, and a fermentation microorganism,Saccharomyces cerevisiae. The maximal ethanol concentration was 4.2 g/L in a batch culture at 15 g/L starch concentration, and 3.6 g/L in a continuous culture fed the same starch concentration. In the fed-batch culture, the maximal ethanol concentration increased more than twofold, compared to the batch culture.

Microbial treatment of kraft pulp wastewater pretreated with ozone

Kraft pulp wastewater was treated with a combination of ozone and activated sludge. The effects of pH on the degradation of lignin and the production of organic acids were examined experimentally in the ozonolysis of wastewater. The strong alkaline condition enhanced not only the degradation of lignin but also the production of organic acids. The dynamic behaviours of microbial growth and substrate consumption were investigated in the biodegradation of organic acids using activated sludge. Maleic acid and oxalic acid in the ozonized wastewater were degraded completely by the activated sludge. The immobilized activated sludge culture using polyurethane foam was effective in degrading organic acids.

Lung inflammation by fungus, Bjerkandera adusta isolated from Asian sand dust (ASD) aerosol and enhancement of ovalbumin-induced lung eosinophilia by ASD and the fungus in mice

BackgroundBjerkandera adusta (B. adusta) is one of the most important etiological fungi associated with chronic cough. However, precise details of the inflammatory response to exposure are not well understood yet. B. adusta was recently identified in Asian sand dust (ASD) aerosol. Therefore, in the present study the exacerbating effects of ASD on B. adusta-induced lung inflammation and B. adusta + ASD on ovalbumin (OVA)-induced murine lung eosinophilia were investigated using experimental mice.MethodsIn order to prepare testing samples, B. adusta obtained from ASD aerosol was inactivated by formalin and ASD collected from the atmosphere was heated to remove toxic organic substances (H-ASD). CD-1 mice were instilled intratracheally with 12 different samples prepared with various combinations of B. adusta, H-ASD, and OVA in a normal saline solution. The lung pathology, cytological profiles in bronchoalveolar lavage fluid (BALF), and the levels of inflammatory cytokines/chemokines in BALF were investigated.ResultsH-ASD aggravated the lung eosinophilia induced by B. adusta alone, which also aggravated the lung eosinophilia induced by OVA. The mixture of OVA, H-ASD, and B. adusta caused serious fibrous thickening of the subepithelial layer, eosinophil infiltration, and proliferation of goblet cells in the airways along with remarkable increases of IL-13, eotaxin, IL-5, and MCP-3 in BALF.ConclusionsThe results of the present study demonstrated that B. adusta isolated from ASD aerosol induces allergic lung diseases. H-ASD enhanced allergic reactions caused by OVA or B. adusta. A mixture of B. adusta, H-ASD, and OVA caused the most remarkable exacerbation to the allergic airway inflammation via remarkable increases of pro-inflammatory mediators.

Immobilization of cross‐linked lipase aggregates onto magnetic beads for enzymatic degradation of polycaprolactone

Candida rugosa lipase was immobilized on amino‐functionalized magnetic supports via cross‐linked enzyme aggregates (CLEA) and used to enhance the enzymatic degradation of polycaprolactone (PCL). The maximum amounts of lipase immobilized on the magnetic beads using glutaraldehyde as a coupling agent were determined to be 33.7 mg/g of beads with an 81% recovery of activity after immobilization. Compared to the free enzyme, the immobilized lipase showed the optimum pH at 1 unit higher (pH 8.0) and also retained its enzymatic activity at higher temperatures. There was 62.9% retention of lipase activity after 30 consecutive reuses, indicating its stability and reusability in aqueous media. Moreover, the immobilized lipase maintained more than 80% of its initial activity during 30 days storage period, while the free lipase lost all under same condition. In addition, the immobilized lipase showed a more than 6‐fold increase in biodegradability over the free lipase when the immobilized lipase was used to degrade PCL in a batch system. Higher thermal and storage stability, as well as good durability after repeated use of the immobilized lipase CLEA, highlights its potential applicability as large scale continuous systems for the enzymatic degradation of PCL. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

Phylogeographic analysis of filterable bacteria with special reference to Rhizobiales strains that occur in cryospheric habitats

Abstract Although the lower size limit of microorganisms was previously believed to be c. 0.2 μm, there is evidence for the existence of microorganisms that can pass through 0.2 μm-pore-size filters called ultramicrobacteria or nanobacteria. However, information on the phylogeny and biogeography of these bacteria is limited. We obtained 53 isolates of 0.2 μm-passable bacteria from 31 samples collected at 26 locations worldwide, including the Arctic Svalbard Islands, deserts, and Maritime Antarctica. Phylogenetic analysis of near full-length 16S rRNA gene sequences revealed that 18 of the 53 isolates were < 97% homologous with previously cultured isolates, representing potentially novel species. Two isolates (order Rhizobiales) (100% identical) collected from Byers Peninsula, Livingston Island in Maritime Antarctica, were closely related (99.8% similarity) to an isolate collected from intertidal sediments in East Antarctica. In addition, the sequence of this Antarctic isolate showed ≥ 97% similarity to 901 sequences derived from known isolates and samples collected at geographically disparate locations under various environmental conditions. Interestingly, among 13 sequences showing ≥ 99% similarity, ten were isolated from cryospheric habitats such as Arctic, Antarctic, and alpine environments. This implies that such Rhizobiales strains occur in the cryospheric regions, however, their abundance and biomass may be scarce depending on the geographic location.

Bioremediation of phenolic compounds having endocrine-disrupting activity using ozone oxidation and activated sludge treatment

The bioremediation of water system contaminated with phenolic compounds having endocrine-disrupting activity,i.e. 2,4-dichlorophenol, 2,4-dichlorophenoxy acetic acid (2,4-D), and 2,4,5-trichlorophenoxy acetic acid (2,4,5-T), was investigated by using ozone oxidation and activated sludge treatment. Ozone oxidation (ozonation time: 30 min) followed by activated sludge treatment (incubation time: 5 days) was an efficient treatment method for the conversion of phenolic compounds in water into carbon dioxide and decreased the value of total organic carbon (TOC) up to about 10% of initial value. Furthermore, 2,4-D was dissolved in water containing salt,i.e. artificial seawater (ASW), and this water was used as model coastal water contaminated with phenolic compounds. The activated sludge treatment (incubation time: 5 days) could consume significantly organic acids produced from 2,4-D in the model costal water by the ozone oxidation (ozonation time: 30 min) and decrease the value of TOC up to about 35% of initial value.