A superoxide dismutase was purified 62-fold in seven steps to homogeneity from Methylobacillus sp. strain SK1, an obligate methanol-oxidizing bacterium, with a yield of 9.6%. The final specific activity was 4,831 units per milligram protein as determined by an assay based on a 50% decrease in the rate of cytochrome c reduction. The molecular weight of the native enzyme was estimated to be 44,000. Sodium dodecyl sulfate gel electrophoresis revealed two identical subunits of molecular weight 23,100. The isoelectric point of the purified enzyme was found to be 4.4. Maximum activity of the enzyme was measured at pH 8. The enzyme was stable at pH range from 6 to 8 and at high temperature. The enzyme showed an absorption peak at 280 nm with a shoulder at 292 nm. Hydrogen peroxide and sodium azide, but not sodium cyanide, was found to inhibit the purified enzyme. The enzyme activity in cell-free extracts prepared from cells grown in manganese-rich medium, however, was not inhibited by hydrogen peroxide but inhibited by sodium azide. The activity in cell extracts from cells grown in iron-rich medium was found to be highly sensitive to hydrogen peroxide and sodium azide. One mol of native enzyme was found to contain 1.1 g-atom of iron and 0.7 g-atom of manganese. The N-terminal amino acid sequence of the purified enzyme was Ala-Tyr-Thr-Leu-Pro-Pro-Leu-Asn-Tyr-Ala-Tyr. The superoxide dismutase of Methylobacillus sp. strain SK1 was found to have antigenic sites identical to those of Methylobacillus glycogenes enzyme. The enzyme, however, shared no antigenic sites with Mycobacterium sp. strain JC1, Methylovorus sp. strain SS1, Methylobacterium sp. strain SY1, and Methylosinus trichosproium enzymes.
International Journal of Industrial Entomology and Biomaterials
/
v.45
no.2
/
pp.93-98
/
2022
To assess the feasibility of silk sericin for non-textile application, the storage stability and biological safety of sericin were examined. It was extracted at 37℃, 70℃, 100℃, and 121℃ for 1, 3, and 5 h to elucidate the effect of extraction condition on the stability and safety of silk sericin. The solubility was increased till approximately 26% with extraction temperature of 121℃ for 1 h. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that the molecular weight distribution depended on the extraction conditions. Extracted sericin displayed typical UV absorption bands upon spectrometric analysis. To examine the reproducibility of its obtained conformation, sericin was extracted thrice and its circular dichroism (CD) spectra was measured each time. Most CD spectra showed reproducibility regardless of temperature and time except under 100℃ extraction condition. The diversity of CD spectrum showed gradual reduction and was finally coincident with extraction time from 1 to 5 h. Notably, sericin has a negative peak of approximately 200 nm attributed to random coil conformation, regardless of extraction condition. However, at the 100℃ extraction condition, sericin showed both bands to be negative bands of approximately 200 and 220 nm, respectively. Sericin was centrifuged to determine the stability of storage conditions. The sericin extracted at 100℃ and 121℃ for 1 h was found to form gel rapidly within 1 h, but at 121℃ condition, the gel fraction was approximately 20% within 1 h which retained its phase regardless of storage time. The gel fraction of sericin extracted at 100℃ for 5 h increased with time, however at the 121℃ for 5 h condition, the gel fraction was measured to be less than 10% regardless of increase in storage time. PetriflimTM AC plates test showed that sericin was safe from aerobic bacteria activity by extraction under high temperature.
Journal of Korean Society of Environmental Engineers
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v.29
no.8
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pp.895-903
/
2007
As for the increasing demanding on the development of direct-ecological landfill monitoring methods, it is needed for critically defining the condition of landfills and their influence on the environment, quantifying the amount of enzymes and bacteria mainly concerned with biochemical reaction in the landfills. This study was thus conducted to understand the fates of contaminants in association with groundwater quality parameters. For the study, groundwater was seasonally sampled from four closed unsanitary landfills(i.e. Cheonan(C), Wonju(W), Nonsan(N), Pyeongtaek(P) sites) in which microbial diversity was simultaneously obtained by 16S rDNA methods. Subsequently, a number of primer sets were prepared for quantifying the specific gene of representative bacteria and the gene of encoding enzymes dominantly found in the landfills. The relationship between water quality parameters and gene quantification were compared based on correlation factors. Correlation between DSR(Sulfate reduction bacteria) gene and BOD(Biochemical Oxygen Demand) was greater than 0.8 while NSR(Nitrification bacteria-Nitrospira sp.) gene and nitrate were related more than 0.9. A stabilization indicator(BOD/COD) and MTOT(Methane Oxidation bacteria), MCR(Methyl coenzyme M reductase), Dde(Dechloromonas denitrificans) genes were correlated over 0.8, but ferric iron and Fli(Ferribacterium limineticm) gene were at the lowest of 0.7. For MTOT, it was at the highest related at 100% over BOD/COD. In addition, anaerobic genes(i.e., nirS-Nitrite reductase, MCR. Dde, DSR) and DO were also related more than 0.8, which showing anaerobic reactions generally dependant upon DO. As demonstrated in the study, molecular biological investigation and water quality parameters are highly co-linked, so that quantitative real-time PCR could be cooperatively used for assessing landfill stabilization in association with the conventional monitoring parameters.
Sulfate reduction and the precipitation of metal sulfides may have great potential to improve water quality of mine effluents in wetland treatment systems. Laboratory experiments using sulfate reducing bacteria (SRB) and limestone to treat effluents from the abandoned Dalsung tungsten-copper mine show that encouraging results, that have been attributed to sulfate reduction. Fe, Al, Cd, Cu and Zn are reduced to below detection limits with $99{\sim}100%$ metal removal rates, Mn is reduced by at least 90% to below 8.0 mg/l, and the pH is raised from 5.12 to 7.60 after 53 days of experiments. In the staged design, laboratory experiments are initiated to determine what would be reasonable substrate materials for remediation of the mine effluents. A substrate mixture containing 70% oak compost and 30% mushroom compost maintains $0.03{\sim}0.04mM$ of lactate, which provides good condition for the SRB granule. A downflow SRB wetland system is proposed as follows : 1) The lower part of the treatment system consists with a 25 cm thick layer of high quality (above 95% of $CaCO_3$) of limestone; 2) The geotextile (geonet) is recommended to be spread on the limestone bed to prevent clogging the limestones with the substrates; 3) The mixture of substrates with 70% oak and 30% spent mushroom composts, and SRB granules overlain on top of the geonet with 25 cm height. The sizes of the passive treatment systems are calculated according to metal loading and permeability criteria : 1) $220m^3$ ($15{\times}15{\times}1m$) for -1 level effluents; 2) $28m^3$ ($5.3{\times}5.3{\times}1m$) for -2 level; and 3) $2700m^3$ ($52{\times}52{\times}1m$) for the -3 level. The -3 level system needs to be broken down into 5 to 15 cells.
This study was performed to investigate applicabilities of sewage sludge and papermill sludge as carbon sources in biologically treating abandoned acidic mine drainage, and effects of limestone. In spite of ranging average 3.3 in influent pH, SRB(Sulfate Reducing Bacteria) was well grown. because effluent pH was maintained by alkalinity a little under for whole stages. TCODcr was high in effluent with washing out in early stage. but its concentration was low with passed time and did not cause the problem of secondary pollution. Removal rate of Mn was remarkably low, but in case of heavy metals such as Fe, the fixation trends showed high as the volume of ${SO_4}^2$ reduction increased. In case of mixing sewage sludge and papermill sludge, when their mixing ratio were 1 : 1 and 2 : 1 respectively, rate of ${SO_4}^2$ reduction and removal rate of heavy metals were high. The mixing ratio 1 : 1 was considered as the most appropriate, because degradability of swewage sludge under short time was higher than that of papermill sludge. As a result of investigating efficiencies of limestone and biological treatment, biological treatment excelled limestone in neutralization and removal rate of heavy metals.
Acid mine drainage (AMD) from abandoned mine sites typically has low pH and contains high level of various heavy metals, aggravating ground- and surface water qualities and neighboring environments. This study investigated removal of heavy metals in a biological treatment system, mainly focusing on the removal by adsorption on a substrate material. Bench-scale batch experiments were performed with a mushroom compost to evaluate the adsorption characteristics of heavy metals leached out from a mine tailing sample and the role of SRB in the overall removal process. In addition, adsorption experiments were perform using an artificial AMD sample containing $Cd^{2+}$, $Cu^{2+}$, $Pb^{2+}$ and $Zn^{2+}$ to assess adsorption capacity of the mushroom compost. The results indicated Mn leached out from mine tailing was not subject to microbial stabilization or adsorption onto mushroom compost while microbially mediated stabilization played an important role in the removal of Zn. Fe leaching significantly increased in the presence of microbes as compared to autoclaved samples, and this was attributed to dissolution of Fe minerals in the mine tailing in a response to the depletion of $Fe^{3+}$ by iron reduction bacteria. Measurement of oxidation reduction potential (ORP) and pH indicated the reactive mixture maintained reducing condition and moderate pH during the reaction. The results of the adsorption experiments involving artificial AMD sample indicated adsorption removal efficiency was greater than 90% at pH 6 condition, but it decreased at pH 3 condition.
Kim, Young-Jae;Hahn, Se-Hyun;Lee, Sang-Hoon;Jang, Ki-Taeg;Kim, Chol-Chul
Journal of the korean academy of Pediatric Dentistry
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v.31
no.4
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pp.605-616
/
2004
Subinhibitory concentrations (sub-MICs) refer to concentrations below minimum inhibitory concentrations (MICs). The antimicrobial agents may be present at relatively high concentration, at least higher than bacterial MIC and thereafter be deserted off a surface and function at sub-MICs, perhaps by interfering with bacterial metabolism. Consequently, the aim of this study was to determine the effects of growth, in the presence of sub-MICs of antimicrobial agents, on the cell surface properties and virulence factors of mutans streptococci and to investigate the efficacy of a chemical approach in vitro. Streptococcus mutans Ingbritt and Streptococcus sobrinus 6715-7 were used. Eight antimicrobial agents (Sanguinaria extract;SG, Chlorhexidine digluconate;CHX, Fluoride;F, Propolis;PP, Hydrogen peroxide;HP, Triclosan;TC, Sodium dodecyl sulfate;SDS Cetylpyridinium chloride; CC) were diluted serially in broth to determine MICs and to compare the growth rate, acid production, hydrophobicity, adhesion activity to saliva coated hydroxyapatite, glucan synthesis and cellular aggregation of experiment groups (in the presence of sub-MICs) with those of control (in the absence of antimicrobial agents). Sub-MICs of antimicrobial agents affected the growth of cells, hydrophobicity, and adhesion of bacteria to saliva coated hydroxyapatite and glucan synthesis. They also resulted in a significant reduction in pH after 12 hours (p<0.05). By cells pretreated with proteinase K, either the aggregation induced by antimicrobial agents was completely inhibited or the aggregation titers were markedly increased. According to the results of the present study, each antimicrobial agent at sub-MICs could affect similar as its known action mechanism and could continually inhibit cariogenic bacteria at such concentrations. Thus, the use of these antimicrobial agents would be one of the effective methods to prevent dental caries.
The study site located in an industrial complex has a Precambrian age gneiss as a bedrock. The poorly-developed, disturbed soils in the study site have loamy-textured surface soil (1 to 2 m) and gravelly sand alluvium subsurface (2 to 6 m) on the top of weathered gneiss bedrock. The depth of the groundwater table was about 3.5 m below ground surface and increased toward down-gradient of the site. The hydraulic conductivity of transmitted zone (gravelly coarse sand) was in the range of 5.0${\times}$10$\^$-2/∼1.85${\times}$10$\^$-1/ cm/sec. The fine sand layer was in the range of 1.5${\times}$10$\^$-3/ to 7.6${\times}$10$\^$-3/ cm/sec. and the reclaimed upper soil layer was less than 10$\^$-4/ cm/sec. Toluene, ethylbenzene, and xylene (TEX) was the major contaminant in the soil and groundwater. The average depth of the soil contamination was about 1.5 m in the gravelly sand alluvium layer. At the depth interval 2.4∼4.8 m, the highest contamination in the soil is located approximately 50 to 70 m from the suspected source areas. The concentration of TEX in the groundwater was highest in the suspected source area and a lesser concentration in the center and southwest parts of the site. The TEX distribution in the groundwater is associated with their distribution in the soil. Microbial isolation showed that Pseudomonas flurescence, Burkholderia cepacia, and Acinetobactor lwoffi were the dominant aerobic bacteria in the contaminated soils. The analytical results of the groundwater indicated that the concentrations of dissolved oxygen (DO), nitrate, and sulfate in the contaminated area were significantly lower than their concentrations in the none-contaminated control area. The results also indicated that groundwater at the contaminated area is under anaerobic condition and sulfate reduction is the predominant terminal electron accepting process. The total attenuation rate was 0.0017 day$\^$-1/ and the estimated first-order degradation rate constant (λ) was 0.0008 day$\^$-1/.
The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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v.10
no.1
/
pp.1-7
/
2005
Tidal flats have been regarded to carry out transformation and removal of land-derived organic matter, and this purifying capability of organic matter by tidal flats is one of very important reasons for their conservation. However, integral biogeochemical studies on production and decomposition of organic matter by benthic microbes in tidal flats have been absent in Korea, although the information is indispensable to quantification of the purifying capability. Our major goals in this multidisciplinary research were to understand major biogeochemical processes and rates mediated by diverse groups of microbes dominating material cycles in the tidal flats, and to assess the contribution of benthic microbes to removal of organic matter and nutrients in the tidal flats. Our study sites were Ganghwa and Incheon north-port tidal flats that had been regarded as naturally well reserved and organically polluted, respectively. Our research group measured over 3 years primary production, biomass and community structure of primary producers, abundance and production of bacteria, enzyme activities, distribution of protozoa and protozoan grazing rates, rates of denitrification and sulfate reduction, early sediment diagenesis, primary production and respiration based on oxygen microelectrode. We analyzed major features of each biogeochemical process and their interactions. The results are compiled in the following articles in this special issue: An (2005), Hwang and Cho (2005), Mok et at. (2005), Na and Lee (2005), Yang et at. (2005), and Yoo and Choi (2005).
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