Electron microscopy study of L1(0)-FePtCu nanoparticles synthesized at 613K.

Finely dispersed hard magnetic L1o-FePtCu nanoparticles with 100 orientation were directly synthesized by RF-sputtering on NaCl substrate at a temperature of 613 K. The maximum coercivity of the particles was 1.4 kOe (at RT). Degrees of atomic long-range order (LRO) for the L1o-FePtCu nanoparticles with different sizes were obtained using nanobeam electron diffraction technique. The decrease of LRO parameter became remarkable when the size became below 8 nm. The coercivity value also decreased with decreasing the particle size. The relation between the LRO parameter decrease and the coercivity decrease with particle size was discussed.

Application of quantitative real-time PCR for quantification of Rhodococcus sp. EH831 in a polyurethane biofilter.

Rhodococcus sp. EH831 is a microbial species that can degrade volatile organic compounds. We optimized a method for monitoring quantitative real-time PCR (qRT-PCR) of EH831 that was incorporated into a polyurethane (PU) biofilter. When the genomic DNA of EH831 was directly extracted from a PU sample with immobilized EH831, the recovery efficiency was very low due to DNA absorption into the PU. DNA amplification during PCR was also inhibited by PU impurities. Therefore, a pre-treatment step was necessary. We successfully recovered cells from the PU by squeezing the matrix, adding sterilized water, and vortexing. The recovery efficiency ranged from 105 to 144%, and there was no statistically significant difference. We designed a novel TaqMan probe for EH831 and demonstrated its high specificity for EH831. The detection range for EH831 was 10(5)-10(11) CFU ml(-1). The method described in this study can be used to investigate the relationship between quantitative analysis of Rhodococcus sp. EH831 and PU biofilter performance.

Inhibitory effects on mushroom tyrosinase by flavones from the stem barks of Morus lhou (S.) Koidz.

Five flavones displaying tyrosinase inhibitory activity were isolated from the stem barks of Morus lhou (S.) Koidz., a cultivated edible plant. The isolated compounds were identified as mormin (1), cyclomorusin (2), morusin (3), kuwanon C (4), and norartocarpetin (5). Mormin (1) was characterized as a new flavone possesing a 3-hydroxymethyl-2-butenyl at C-3. The inhibitory potencies of these flavonoids toward monophenolase activity of mushroom tyrosinase were investigated. The IC50 values of compounds 1-5 for monophenolase activity were determined to be 0.088, 0.092, 0.250, 0.135 mM, and 1.2 microM, respectively. Mormin (1), cyclomorusin (2), kuwanon C (4) and norartocarpetin (5) exhibited competitive inhibition characteristics. Interestingly norartocarpetin (5) showed a time-dependent inhibition against oxidation of L-tyrosine: it also operated under the enzyme isomerization model (k5 = 0.8424 min(-1), k6 = 0.0576 min(-1), K(app)(i) = 1.354 microM).

Batch and repeated batch production of L (+)-lactic acid by Enterococcus faecalis RKY1 using wood hydrolyzate and corn steep liquor.

Lactic acid production was investigated for batch and repeated batch cultures of Enterococcus faecalis RKY1, using wood hydrolyzate and corn steep liquor. When wood hydrolyzate (equivalent to 50 g l(-1) glucose) supplemented with 15-60 g l(-1) corn steep liquor was used as a raw material for fermentation, up to 48.6 g l(-1) of lactic acid was produced with, volumetric productivities ranging between 0.8 and 1.4 g l(-1 )h(-1). When a medium containing wood hydrolyzate and 15 g l(-1) corn steep liquor was supplemented with 1.5 g l(-1) yeast extract, we observed 1.9-fold and 1.6-fold increases in lactic acid productivity and cell growth, respectively. In this case, the nitrogen source cost for producing 1 kg lactic acid can be reduced to 23% of that for fermentation from wood hydrolyzate using 15 g l(-1) yeast extract as a single nitrogen source. In addition, lactic acid productivity could be maximized by conducting a cell-recycle repeated batch culture of E. faecalis RKY1. The maximum productivity for this process was determined to be 4.0 g l(-1 )h(-1).

Microbial characterization of toluene-degrading denitrifying consortia obtained from terrestrial and marine ecosystems.

The degradation characteristics of toluene coupled to nitrate reduction were investigated in enrichment culture and the microbial communities of toluene-degrading denitrifying consortia were characterized by denaturing gradient gel electrophoresis (DGGE) technique. Anaerobic nitrate-reducing bacteria were enriched from oil-contaminated soil samples collected from terrestrial (rice field) and marine (tidal flat) ecosystems. Enriched consortia degraded toluene in the presence of nitrate as a terminal electron acceptor. The degradation rate of toluene was affected by the initial substrate concentration and co-existence of other hydrocarbons. The types of toluene-degrading denitrifying consortia depended on the type of ecosystem. The clone RS-7 obtained from the enriched consortium of the rice field was most closely related to a toluene-degrading and denitrifying bacterium, Azoarcus denitrificians (A. tolulyticus sp. nov.). The clone TS-11 detected in the tidal flat enriched consortium was affiliated to Thauera sp. strain S2 (T. aminoaromatica sp. nov.) that was able to degrade toluene under denitrifying conditions. This indicates that environmental factors greatly influence microbial communities obtained from terrestrial (rice field) and marine (tidal flat) ecosystems.

Leaching characteristics of heavy metals from sewage sludge by Acidithiobacillus thiooxidans MET.

An acidophilic, sulfur-oxidizing Acidithiobacillus thiooxidans MET bacterium was isolated from anaerobically digested, dewatered sewage sludge. This bacterium showed sulfur-oxidizing ability at both acidic and neutral conditions, and allowed metal leaching even at a high (130 g L(-1)) sludge solids concentration. We found that low metal leaching efficiency at high solids concentration was mainly due to an increase in buffering capacity resulting in retardation of pH reduction. Therefore, metal leaching was mainly influenced not by sludge solids concentration, but by the pH (or sulfate concentration per unit sludge mass) of the sludge solutions. The relationship between the pH of the sludge solution and the efficiency of metal leaching was obtained by quantitatively investigating the effect of pH reduction or the amount of sulfate produced per unit sludge mass on leaching of each metal. Furthermore, the relationship between total metal content in the sludge and metal leached to the solution was obtained for each metal. Such a relationship allowed estimation of leachable metal at various amounts of total metal content in sludge.

Perifusion model for detoxification of drugs in a bioartificial liver.

Detoxification of a drug in a bioartificial liver (BAL) during an in vitro experiment was theoretically carried out based on a perifusion model. The detoxification capacity assay, the rates of disappearance of the chemicals such as flow-limited and enzyme-limited drugs in the BAL system could be defined from models of hepatic perfusion-elimination relationships. When the flow-limited drug administrated under a quasi-equilibrium condition, a two-compartment model for the concentration behavior of the drug was introduced and compared with a one-compartment model. For both models, equations involving hepatic drug clearance and various pharmacokinetic parameters were derived under initial bolus loading and constant-rate infusion plus bolus loading conditions. The concentration of enzyme-limited drug in the BAL decreased linearly with time in contrast with the concentration profile of the flow-limited drug followed by exponential functions. The perifusion model offers a quantitative understanding of the elimination kinetics of chemicals such as flow-limited and enzyme-limited drugs in a bioartificial liver and a comparison between the BAL and human liver.

Hydrogen sulfide effects on ammonia removal by a biofilter seeded with earthworm casts.

Ammonia (NH3) removal efficencies were evaluated when hydrogen sulfide (H2S) and NH3 in binary mixture gases were supplied to a ceramic biofilter seeded with earthworm (Lumbricus terrestris) casts. The effect of inlet H2S concentration and space velocity (SV) on the removal of NH3 was investigated after the acclimation of the biofilter with NH3 gas. When NH3 was singly supplied to the biofilter, NH3 removal was maintained at almost 100% until inlet NH3 concentration was increased up to 600 microL L(-1) and SV up to 330 h(-1), at which the elimination capacity of NH3 was 148 g N m(-3) h(-1). When H2S was supplied simultaneously, however, the accumulation of toxic sulfide ions showed dual effects on NH3 removal efficiencies. First, no effects were observed at inlet H2S loading below 60 g S m(-3) h(-1); however, inhibition by H2S at higher loading was observed above 60 g S m(-3) h(-1). The point at which loading achieved a maximum of more than 99% NH3 removal efficiency was 139 g N m(-3) h(-1), when inlet H2S concentration was held under 100 microL L(-1), but it dropped to 76 and 30 g N m(-3) h(-1) when the inlet H2S concentration increased to 220 and 460 microL L(-1), respectively. The critical points of inlet H2S loading that guaranteed over 99% NH3 removal were determined as 100, 100, 60, and 40 g S m(-3) h(-1) at inlet NH3 concentrations of 100, 200, 400, and 600 microL L(-1), respectively. Inlet NH3 loading had synergic effects of increasing the inhibition of inlet H2S loading on the NH3 removability of the biofilter.

Production of a desulfurization biocatalyst by two-stage fermentation and its application for the treatment of model and diesel oils.

For the production of oil-desulfurizing biocatalyst, a two-stage fermentation strategy was adopted, in which the cell growth stage and desulfurization activity induction stage were separated. Sucrose was found to be the optimal carbon source for the growth of Gordonia nitida CYKS1. Magnesium sulfate was selected to be the sulfur source in the cell growth stage. The optimal ranges of sucrose and magnesium sulfate were 10-50 and 1-2.5 g x L(-1), respectively. Such a broad optimal concentration of sucrose made the fed-batch culture easy, while the sucrose concentration was maintained between 10-20 g x L(-1) in the actual operation. As a result, 92.6 g x L(-1) of cell mass was acquired by 120 h of fed-batch culture. This cell mass was over three times higher than a previously reported result, though the strain used was different. The desulfurization activity of the harvested cells from the first stage culture was induced by batch cultivation with dibenzothiophene as the sole sulfur source. The optimal induction time was found to be about 4 h. The resting-cell biocatalyst made from the induced cells was applied for the deep desulfurization of a diesel oil. It was observed that the sulfur content of the diesel oil decreased from 250 mg-sulfur x L-oil(-1) to as low as 61 mg-sulfur x L-oil(-1) in 20 h. It implied that the biocatalyst developed in this study had a good potential to be applied to a deep desulfurization process to produce ultra-low-sulfur fuel oils.

Characteristics and glycerol metabolism of fumarate-reducing Enterococcus faecalis RKY1.

Enterococcus faecalis RKY1, which converts fumarate to succinate with a high yield, was identified on the basis of a phylogenetic analysis of the 16S rDNA gene sequence. The strain was incubated at 38 degrees C for 18 h to examine the possible diversion of glucose or glycerol fermentation by fumarate. The products of glucose and glycerol fermentation with fumarate were quite different from those of normal fermentation, which ultimately produces lactate, in that mainly succinate is produced. Metabolic pathway stoichiometry was used to analyze the oxidation of glycerol to succinate by Enterococcus faecalis RKY1. The stoichiometric relationship between glycerol and fumarate was used as a guideline to accumulate succinate more efficiently.

Characterization of bioconversion of fumarate to succinate by alginate immobilized Enterococcus faecalis RKY1.

In this study, the immobilization characteristics of Enterococcus faecalis RKY1 for succinate production were examined. At first, three natural polymers--agar, kappa-carrageenan, and sodium alginate--were tried as immobilizing matrices. Among these, sodium alginate was selected as the best gel for immobilization of E. faecalis RKY1. Efficient conditions for immobilization were established to be with a 2% (w/v) sodium alginate solution and 2-mm-diameter bead. The bioconversion characteristics of the immobilized cells at various pH values and temperatures were examined and compared with those of free cells. The optimum pH and temperature of the immobilized cells were the same as for free cells, 7.0 and 38 degrees C respectively, but the conversion ratio was higher by immobilization for all the other pH and temperature conditions tested. When the seed volume of the immobilized cells was adjusted to 10% (v/v), 30 g/L of fumarate was completely converted to succinate (0.973 g/g conversion ratio) after 12 h. In addition, the immobilized cells maintained a conversion ratio of >0.95 g/g during 4 wk of storage at 4 degrees C in a 2% (w/v) CaCl2 solution. In repetitive bioconversion experiments, the activity of the immobilized cells decreased linearly according to the number of times of reuse.

Utilization of swine wastewater as a feedstock for the production of polyhydroxyalkanoates by Azotobacter vinelandii UWD.

Azotobacter vinelandii UWD produced 0.69 g.l(-1) poly(hydroxybutyrate-co-hydroxyvalerate, PHBV) with 7.9 mol% hydroxyvalerate (HV) from twofold-diluted swine wastewater (SW). When supplemented with 20 g.l(-1) glucose, twofold-diluted SW increased copolymer production by 8.6 times. When three organic acids (acetate, propionate and butyrate) present in SW were supplemented with 20 g.l(-1) glucose, PHBV production was comparable (5.5 g.l(-1)) to that in the case of using twofold-diluted SW supplemented with 20 g.l(-1) glucose. However, the HV level (1.1-1.3 mol%) was very low. The component in SW contributing to copolymer production was found to be valerate. By 20 mM valerate 0.2 g.l(-1) PHBV with 44.6 mol% HV was produced. With 30 g.l(-1) glucose supplementation, 4.0 g.l(-1) PHBV with 22 mol% HV was produced. The optimal ratios of carbon to phosphorus (C : P) and to nitrogen (C : N) were 165 : 1 and 22 : 1, respectively.

Leaching of Mn, Co, and Ni from manganese nodules using an anaerobic bioleaching method.

An anaerobic bioleaching of a manganese nodule by anaerobic Mn-reducing bacteria was evaluated for the leaching of metals, Mn, Co, and Ni. Insoluble Mn4+ in the nodule could be reduced to soluble Mn2+ by dissimilatory Mn-reducing bacteria that use a carbon source and Mn4+ as an electron donor and acceptor, respectively. As a result of the Mn reduction, Co and Ni could be leached from the loosed Mn matrix. Leaching experiments were carried out to optimize various process parameters, such as inoculation, pH, temperature, mineral salts, and particle size of the nodule used. The leaching efficiencies of Mn, Co, and Ni increased from 18, 7, and 10% to 77, 70, and 75%, respectively by the inoculation of the Mn-reducing enrichment culture broth. Metals could be efficiently recovered from the nodule in the ranges of pH from 5.0 to 6.5 and temperature from 30 to 45 degrees C by anaerobic bioleaching. External addition of mineral salts was not necessary for Mn, Co, and Ni leaching from the nodule. The optimum ratio of nodule to glucose was 0.1 (w/w). To obtain a leaching efficiency above 70%, the particle size of the nodules must be less than 0.6 mm.

Simultaneous determination of delta-aminolevulinic acid, porphobilinogen, levulinic acid and glycine in culture broth by capillary electrophoresis.

Capillary electrophoretic simultaneous determination of a mixture containing delta-aminolevulinic acid, porphobilinogen, levulinic acid and glycine was investigated. With increases in the sodium tetraborate buffer concentration (5-70 mM), resolution of the four components was improved, but the migration time was increased. Alternatively, with increases in the applied voltage (5-22.5 kV), a shortened migration time was seen but this adversely affected resolution. The components were separated with high resolution by using a fused-silica capillary column (75 cm x 75 microm I.D.) filled with 30 mM sodium tetraborate buffer (pH 9.3-9.4) under the applied voltage of 20 kV (constant voltage mode). When the established method was applied to the culture broth of Rhodopseudomonas sphaeroides, a photosynthetic bacterium, the four components mentioned above were separated with good resolution. Furthermore, the use of this method would provide a fast, sensitive and specific method for monitoring the administration of delta-aminolevulinic acid in photodynamic cancer therapy, for the measurement of delta-aminolevulinic acid dehydratase activity in erythrocytes, and for testing the delta-aminolevulinic acid assay and for impurities in drug formulation.

Recovery of poly(3-hydroxybutyrate) from coagulated Ralstonia eutropha using a chemical digestion method.

For economic recovery of poly(3-hydroxybutyrate) (PHB) from culture broths of Ralstonia eutropha containing PHB, Al-based and Fe-based coagulants were used in the pretreatment step. The coagulated cells were then separated by centrifugation, and PHB was extracted by chemical digestion with a sodium hypochlorite/chloroform dispersion solution. The practical upper limits of dosage were found to be 1, 500 mg-Al/L and 1,000 mg-Fe/L, respectively, for Al- and Fe-based coagulants. When the harvested cells were treated with a 50% sodium hypochlorite/chloroform dispersion solution, PHB recovery and purity were 90-94% and 98-99%, respectively. The influence of the use of coagulants on the PHB recovery process was found to be insignificant. Despite the residual Al and Fe in the recovered PHB (less than 450 mg-Al/kg-PHB and 750 mg-Fe/kg-PHB, respectively), no detectable amounts of Al and Fe were leached from films made of the recovered PHB under acidic conditions. The use of Fe-based coagulants is less recommended because the Fe impurity can cause an unwanted colorization problem in the final product.

Separation of Alcaligenes eutrophus cells containing Poly(3-hydroxybutyrate) from fermentation broth with pretreatment using Al- and Fe-based coagulants.

Alcaligenes eutrophus containing intracellular poly(3-hydroxybutyrate) was recovered from fermentation broth by centrifugation and filtration after pretreatment with Al- and Fe-based coagulants. Coagulation efficiency was largely affected by pH, and the optimum pH's for cell recovery were about 4.6-5.6 for the Al-based coagulants and about 5-8 for the Fe-based coagulants. Ammonium ions that combined with metals to form complex compounds increased the coagulant requirement, and the additional requirement of coagulant was found to be proportional to the ammonium concentration. In addition, various ligands in addition to ammonium ions contained in the culture medium interfered with the coagulation reaction and increased the coagulant requirement also. The coagulant requirement increased with the cell concentration regardless of coagulant type. The polymeric coagulants such as PACS, Hi-PAX, and Ferix-3 were more effective than nonpolymeric coagulants of aluminum sulfate and ferrous sulfate. The optimum dosages of the coagulants tested were determined over a broad range of cell concentration of 20.5-210 g/L. It was observed that the energy requirement for centrifugation could be greatly reduced with cell coagulation.

Desulfurization of light gas oil in immobilized-cell systems of Gordona sp. CYKS1 and Nocardia sp. CYKS2.

Desulfurizations of a model oil (hexadecane containing dibenzothiophene (DBT)) and a diesel oil by immobilized DBT-desulfurizing bacterial strains, Gordona sp. CYKS1 and Nocardia sp. CYKS2, were carried out. Celite bead was used as a biosupport for cell immobilization. Seven-eight cycles of repeated-batch desulfurization were conducted for each strain. Each batch reaction was carried out for 24 h. In the case of model oil treatment with strain CYKS1, about 4.0 mM of DBT in hexadecane (0.13 g sulfur l(oil)(-1)) was desulfurized during the first batch, while 0.25 g sulfur l(oil)(-1) during the final eighth batch. The mean desulfurization rate increased from 0.24 for the first batch to 0.48 mg sulfur l(dispersion)(-1) h(-1) for the final batch. The sulfur content in the light gas oil was decreased from 3 to 2.1 g l(oil)(-1) by strain CYKS1 in the first batch. The mean desulfurization rate was 1.81 mg sulfur l(dispersion)(-1) h(-1), which decreased slightly when the batch reaction was repeated. No significant changes in desulfurization rate were observed with strain CYKS2 when the batch reaction was repeated. When the immobilized cells were stored at 4 degrees C in 0.1 M phosphate buffer (pH 7.0) for 10 days, the residual desulfurization activity was about 50 approximately 70% of the initial value.

Biological deodorization of hydrogen sulfide using porous lava as a carrier of Thiobacillus thiooxidans.

Biological deodorization of hydrogen sulfide (H2S) was studied using porous lava as a carrier of Thiobacillus thiooxidans in a laboratory-scale biofilter. Three different samples of porous lava, A, B, and C, which were obtained from Cheju Island in Korea, were used. The water-holding capacities of samples A, B and C were 0.38, 0.25, and 0.47 g-H2O/g-lava, respectively. The pHs and densities of the lava samples ranged from 8.25-9.24 and 920-1190 kg/m3, respectively. The buffering capacities, expressed as the amount of sulfate added to lower the pH to 4, were 60 g-SO4(2-)/kg-lava for sample A, 50 g-SO4(2-)/kg-lava for B, and 90 g-SO4(2-)/kg-lava for C. To investigate the removal characteristics of H2S by the lava biofilters, T. thiooxidans was immobilized on the lava samples. Biofilters A and C showed a removal capacity of 428 g-S.m(-3).h(-1) when H2S was supplied with 428 g-S.m(-3).h(-1) of inlet load at a space velocity (SV) of 300 h(-1). At the same inlet load and SV, the removal capacity of biofilter B was 396 g-S.m(-3).h(-1). The H2S critical loads of biofilters A, B and C at a SV of 400 h(-1) were 396, 157 and 342 g-S.m(-3).h(-1), respectively. It is suggested that natural, porous lava is a promising candidate as a carrier of microorganisms in biofiltration.

Bioconversion of fumarate to succinate using glycerol as a carbon source.

In this study, a facultative bacterium that converts fumarate to succinate at a high yield was isolated. The yield of bioconversion was enhanced about 1.2 times by addition of glucose into culture medium at an initial concentration of 6 g/L. When the initial cell density was high (2 g/L), the succinate produced at pH 7.0 for initial fumarate concentrations of 30, 50, 80, and 100 g/L were 29.3, 40.9, 63.6, and 82.5 g/L, respectively, showing an increase with the initial fumarate concentration. The high yield of 96.8%/mole of fumarate in just 4 h was obtained at the initial fumarate concentration of 30 g/L. Comparing these values to those obtained with low cell culture (0.2 g/L), we found that the amount of succinate produced was similar, but the production rate in the high cell culture was about three times higher than was the case in the low cell culture. This strain converted fumarate to succinate at a rate of 3.5 g/L.h under the sparge of CO2.

Microbial removal of Fe(III) impurities from clay using dissimilatory iron reducers.

Fe(III) impurities, which detract refractoriness and whiteness from porcelain and pottery, could be biologically removed from low-quality clay by indigenous dissimilatory Fe(III)-reducing microorganisms. Insoluble Fe(III) in clay particles was leached out as soluble Fe(II), and the Fe(III) reduction reaction was coupled to the oxidation of sugars such as glucose, maltose and sucrose. A maximum removal of 44-45% was obtained when the relative amount of sugar was 5% (w/w; sugar/clay). By the microbial treatment, the whiteness of the clay was increased from 63.20 to 79.64, whereas the redness was clearly decreased from 13.47 to 3.55.